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Marchuk, Douglas Alan

Overview:

Vascular Morphogenesis: A Human Genetics Approach

Advances in our understanding of fundamental biological events can often be made by the analysis of defects manifested in inherited diseases. The genes responsible for these genetic syndromes often encode proteins that act at critical points of the pathways that control fundamental biological processes such as cell division, differentiation, and cell death. This approach has lead to the discovery of novel gene products and/or biochemical pathways involved in disease, genes that in turn play a fundamental role in normal biological processes.

My laboratory has taken this genetic approach for the study of angiogenesis, beginning with Mendelian disorders of vascular dysplasia and progressing to more complex vascular phenotypes. Our objectives are twofold: (1) to gain specific knowledge of the pathology observed in these disorders, and (2) to provide basic knowledge on the role of these genes and gene products in angiogenesis. The first step is to identify the genetic loci that underlie these syndromes. These mapping and positional cloning endeavors become the basis for future molecular biological studies on the role of the mutant gene products in the pathology of the disease, and the role of the normal proteins in vascular development. In order to investigate disease mechanism and pathogenesis, we then create an appropriate transgenic or knockout mouse as an animal model of the disease. The animal model serves both as a tool to further understand the pathophysiology of the disease, and a more tractable system to begin to identify other factors (genetic and environmental) that may alter the phenotype. Coming full circle, we can determine if the factors identified in the animal model also modify the clinical phenotype in the human disease. We currently have four projects that utilize this approach, and a new initiative that begins with a mouse model of hypertrophic cardiomyopathy.

Project 1. Molecular Genetics of Arterio-Venous Communication: Hereditary Hemorrhagic Telangiectasia: Hereditary Hemorrhagic Telangiectasia (HHT or Osler-Weber-Rendu disease) is an autosomal dominant disorder characterized by hemorrhagic stroke, gastrointestinal bleeding, and other vascular pathology. The clinical features result from the development of focal vascular malformations characterized by direct arteriovenous shunts with a loss of the capillary beds. The pathology of this disorder suggests a critical role for the HHT gene(s) in vascular development and angiogenesis of the capillary bed. However, until our work on this disorder, the nature of the molecular defect remained unknown. We have shown that HHT is actually a group of related disorders with overlapping but distinct phenotypes and genetic etiologies (McAllister et al., 1994a; Porteous et al., 1994; Berg et al., 1996). We established genetic linkage at two distinct loci for HHT, one on chromosome 9q33 (McDonald et al., 1994) and the other on 12q13 (Johnson DW, et al., 1995). We subsequently identified the gene for HHT 1 as endoglin (McAllister et al., 1994b), a transforming growth factor-ÿ(TGF-ÿÿ) binding protein of endothelial cells, and the HHT 2 locus as the activin-like kinase receptor, ALK-1, which has sequence homology to type I TGF-ÿÿ receptors (Johnson et al., 1996). Expression analyses of the mutant allele for endoglin (McAllister et al., 1995; Gallione et al., 1998) and for ALK-1 (Berg et al., 1997; Klaus et al., 1998) has shown that most alleles lead to unstable message or reduced levels of protein, suggesting inherited haploinsufficiency. These data suggest a critical role for the TGF-ÿÿ signal transduction pathway in the pathology of this disease, and more significantly, in the development and/or repair of blood vessels.

Our research has more recently involved a combination of genetic and molecular biological approaches to further study the role of ALK-1 and endoglin in cardiovascular disease and angiogenesis. We are investigating whether sequence polymorphisms in the endoglin and/or ALK-1 genes are risk factors for cerebrovascular disease in the general population with retrospective case-control association studies. We have already shown that one sequence polymorphism in endoglin is associated with increased risk for hemorrhagic stroke (Alberts et al., 1997), an observation which has now been replicated by others in the Japanese population (Takenaka et al., 1999). We are currently attempting to understand how this intronic polymorphism may affect RNA splicing or message stability. Three endoglin coding-polymorphisms are also under investigation, as the intronic polymorphism may be in linkage disequilibrium with one of these.

Although both endoglin and ALK-1 share sequence homology to the TGF-ÿ family of receptors, little is known about their precise role in signaling. Due to the lack of a signaling assay for ALK-1, there is no evidence other than sequence homology that ALK-1 is a TGF-ÿ receptor. We created a signaling assay for ALK-1 by constructing a chimeric receptor consisting of the extracellular ligand-binding domain of ALK-1 fused to the kinase domain of the TGF-ÿ receptor TÿRI, which can activate a reporter gene driven by the PAI-1 promoter. Using this chimera, we have demonstrated ligand specific activation for TGF-ÿ1, -ÿÿ3 (but not -ÿÿ2), and an additional uncharacterized ligand present in serum (Lux et al., 1999). Significantly, this ligand specificity for the TGF-ÿ isoforms parallels that of endoglin. We have also shown that ALK-1 and endoglin can be immunoprecipitated together in a receptor complex (Lux et al., 1999), suggesting that HHT is a result of altered signaling via an endothelial-specific TGF-ÿ receptor complex.

We have used this signaling assay to attempt to identify the novel ligand present in serum. Results with all available mammalian TGF-ÿ family members were negative (Lux et al., 1999). The Drosophila SAX gene encodes a receptor which may be the most similar in this species to mammalian ALK-1. We have now shown that the Drosophila SAX ligand screw (SCW) activates the ALK-1 chimera in our assay (Lux, Arora, and Marchuk, unpublished). The reciprocal experiment, performed in the laboratory of our collaborator Kavita Arora (UC, Irvine) suggests that mammalian ALK-1 can partially substitute for SCW in Drosophila development. The combined data suggest that ALK-1 signaling may involve a novel TGF-ÿ related ligand, and that a novel biochemical pathway involved in fundamental angiogenic processes awaits identification. Attempts to clone the mammalian homologue of the SCW ligand are in progress, using a variety of bioinformatic, molecular and biochemical approaches. Using our signaling assay, we are attempting to identify the downstream effectors of this pathway, as well as characterize the changes in gene expression due to signaling through these receptors.

We also wish to create animal models for HHT1 and 2. We now have ALK-1 and endoglin knockout mice. Although in both cases the mutant homozygotes show embryonic lethality, we have recently identified gastrointestinal vascular malformations (telangiectasias) in the ALK-1 heterozygotes (Yu, McLendon, and Marchuk, unpublished). The mice have become a critical resource for future studies on HHT, as will be outlined in the section on Future Directions.

Project 2. Molecular Genetics of Venous Angiogenesis: Familial Venous Malformations: Venous malformations are composed of large vascular lumens lined by a single-layered flat mature endothelium, which are thus prone to hemorrhage. The cutaneous and visceral lesions occur as single or multiple lesions with a nodular or tumor-like appearance. The presence of a few families exhibiting autosomal dominant inheritance of these vascular lesions suggested that genetic linkage analysis and positional cloning could be used to identify the responsible gene(s). We confirmed an earlier report by the laboratories of Bjorn Olsen and Matt Warman of linkage to chromosome 9p21 for autosomal dominant venous malformations (Gallione et al., 1995). We subsequently mapped the gene for the tie-2 receptor kinase to the candidate interval, and in collaboration with this group, identified an identical mutation in the kinase domain of tie-2 for two large kindreds (Vikkula et al., 1996). In our laboratory, we subsequently identified a second missense mutations in tie-2 which causes FVM, and have shown that both known mutations constitutively activate the receptor (Calvert et al., 1999). Although the tie-2/angiopoietin pathway has been implicated in vasculogenesis and embryonic angiogenesis from knockout studies in the mouse, our work suggests that this pathway has a role in the maintenance of proper vascular structure extending into adulthood. We speculate that this pathway is involved in endothelial-smooth muscle association based on the anatomy of the vascular lesion.

We are now creating a transgenic mouse model of this disorder, by expressing both tie-2 mutations under the control of its own promoter. This mouse model will be most useful for our understanding of the initiating event in lesion formation in these families, since although the phenotype is expressed in the heterozygous state, there appears to be no global vascular pathology outside of the vascular lesions themselves.

We have also identified 5 additional families with a clinical presentation similar to our tie-2 venous malformation families, but which exclude the tie-2 locus by linkage analysis. In some families the lesions were classified as glomangiomas, a benign tumor derived from the glomus cells (smooth muscle derivative) of the vasculature. We have recently shown that these families map to chromosome 1p (Calvert et al. submitted), and are actively searching for mutations in genes mapping to the critical region.

Project 3. Molecular Genetics of Cerebrovascular Angiogenesis: Cerebral Cavernous Malformations: Cerebral cavernous malformations (CCM) are congenital vascular anomalies of the brain comprising focal, thin-walled, grossly dilated vascular spaces. The lesions are responsible for significant neurologic disability, in particular, intractable migraine, seizures, and hemorrhagic stroke. Autosomal dominant forms of CCM have been described and we and others have shown that a gene for CCM (CCM1) maps to chromosome 7q (Marchuk et al., 1995). Taking advantage of a shared disease haplotype in Mexican-American families, we were able to narrow the critical region to under 500 kb (Johnson et al., 1995c; and unpublished data). We have now identified the CCM1 gene as KRIT1, a recently discovered binding partner of the Krev-1/rap1a tumor suppressor gene (Sahoo et al., in press). A common mutation in most Hispanic families (16 of 21 families analyzed) confirms the founder effect in this population. Other Hispanic and non-Hispanic families harbor different mutations, all of which appear to be null alleles. These data show the strength of this genetic approach to cardiovascular disease, as the Krev-1/rap1a signaling pathway, although implicated in cancer (e.g. Tuberous sclerosis), has not previously been shown to be required for angiogenesis, nor has it been previously implicated in any cardiovascular pathology. We are currently working to create a mouse model of CCM1, as well as to characterize the role of this biochemical pathway in the pathology of hemorrhagic stroke associated with CCM. Mutation analysis is already underway to determine the involvement of KRIT1 mutations/polymorphisms in common (non-hereditary) cerebral cavernous malformations.

Project 4. Molecular Genetics of Capillary Angiogenesis: Hemangiomas: Our work on the autosomal dominant vascular disorders has identified genes involved in the regulation of vascular growth. However, all of these germline mutations are compatible with normal vascular development. We also wish to identify novel genes that might be essential for vasculogenesis and embryonic angiogenesis. Such genes can in principle be identified using human phenotypes, by searching for somatic mutations underlying non-inherited vascular anomalies. We are using this approach to identify the gene(s) underlying the most common tumor (of any kind) in infancy - hemangiomas.
Hemangiomas are benign tumors consisting primarily of proliferating capillaries, which often occur as an elevated purple or red spot on the skin. Hemangiomas usually develop shortly after birth, but are self-limiting and go through a characteristic two-staged process of growth and regression. The rapid proliferation stage suggests an uncontrolled stage of angiogenesis. Our hypothesis is that hemangiomas arise from an early somatic mutation within a critical gene for capillary angiogenesis. Due to the localized nature of the tumor, our prediction is that the mutated gene products will be acting in a cell-autonomous nature, with the tumor resulting from a clonal expansion cell containing the original mutation. Using a clonality assay based on non-random X-chromosome inactivation, we have shown that most proliferative hemangiomas appear mono-clonal (Walter and Marchuk, in preparation). We have also shown significant loss of heterozygosity for markers on chromosome 5q (Berg et al., in press), in the same region that we have identified a genetic susceptibility locus (see below). We are currently searching for mutations in the genes involved in the vascular endothelial growth factor (VEGF) pathway.

Although the great majority of hemangiomas occur sporadically, we have identified a number of families displaying autosomal dominant segregation of childhood hemangiomas (Blei et al., 1998). Using these families to identify genetic loci that may predispose children to hemangioma development, we have found linkage to a region on distal chromosome 5q (Walter et al., 1999). We hope that these independent lines of analysis will converge as we discover that some of these genetic loci are involved in both familial and sporadic cases, the mutations being inherited in the familial cases and acquired somatically in the sporadic cases.

Project 5. Molecular Genetics of Hypertrophic Cardiomyopathy: A Mouse Model of Heart Failure: Heart failure is a common cause of mortality in the United States. It is the final outcome of a variety of conditions, both primary and secondary, that affect the heart. Myocardial hypertrophy and the progression to heart failure are highly heterogeneous and are the result of a combination of environmental and genetic factors. The genetic factors in particular have been recalcitrant to identification.

Mice with cardiac-specific over-expression of the calsequestrin (CSQ) gene develop a severe form of hypertrophic cardiomyopathy that serves as an appropriate mouse model of heart failure. Intriguingly, the extent of cardiomyopathy and the progression to heart failure are highly strain dependent, suggesting the existence of modifying genes in each strain. Using a backcross strategy and quantitative trait locus (QTL) mapping, we have identified three genetic loci that affect the progression of disease in this mouse model (Carlson, Suzuki, Marchuk and Rockman, in preparation). A gene on mouse chromosome 2 strongly affects the progression to heart failure, accounting for 37% of the phenotypic variation in the disease outcome. A locus on chromosome 4 shows a slightly lower effect on disease outcome. In addition, another locus on chromosome 3 strongly affects the extent of cardiomyopathy, as measured by % fractional shortening of the heart. We are in the process of refining the map positions of these three loci, with the goal of the identification of these genes affecting cardiomyopathy and heart failure in this mouse model. Polymorphic variants in these same genes in the human population may be the elusive genetic risk factors for heart failure. Once identified, these will be tested in case-control association studies with appropriate patient populations.

Positions:

James B. Duke Professor of Molecular Genetics and Microbiology

Molecular Genetics and Microbiology
School of Medicine

Professor of Molecular Genetics and Microbiology

Molecular Genetics and Microbiology
School of Medicine

Vice Chair, Department of Molecular Genetics & Microbiology

Molecular Genetics and Microbiology
School of Medicine

Director, Duke University Program in Genetics and Genomics

Molecular Genetics and Microbiology
School of Medicine

Director, Center for Experimental Genetics

Molecular Genetics and Microbiology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

Ph.D. 1985

Ph.D. — University of Chicago

Grants:

Multidisciplinary Heart and Vascular Diseases

Administered By
Medicine, Cardiology
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 1975
End Date
March 31, 2021

Signaling Aberrations and Cerebral Cavernous Malformation Pathogenesis

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 01, 2015
End Date
June 30, 2020

Genetics Training Grant

Administered By
Basic Science Departments
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
September 01, 1979
End Date
June 30, 2020

Organization and Function of Cellular Structure

Administered By
Basic Science Departments
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 1975
End Date
June 30, 2020

Functional Characterization of the GNAQ somatic mutation causing Sturge Weber syndrome

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
April 01, 2015
End Date
March 31, 2018

Development of BA-1049 for treatment of cerebral cavernous malformations

Administered By
Molecular Genetics and Microbiology
AwardedBy
BioAxone Biosciences, Inc.
Role
Principal Investigator
Start Date
August 25, 2016
End Date
July 31, 2017

Brain Vasular Malformation Consortium: Predictors of clinical course

Administered By
Molecular Genetics and Microbiology
AwardedBy
The Regents of the University of California
Role
Principal Investigator
Start Date
September 30, 2014
End Date
July 31, 2017

Duke Training Grant in Nephrology

Administered By
Medicine, Nephrology
AwardedBy
National Institutes of Health
Role
Preceptor
Start Date
September 20, 1995
End Date
June 30, 2017

ROCK INHIBITION AS THERAPY FOR CEREBRAL CAVERNOUS MALFORMATION

Administered By
Molecular Genetics and Microbiology
AwardedBy
University of Chicago
Role
Principal Investigator
Start Date
June 01, 2016
End Date
May 31, 2017

Institutional Training Grant in Pediatric Cardiology

Administered By
Pediatrics, Cardiology
AwardedBy
National Institutes of Health
Role
Faculty Member
Start Date
April 01, 2009
End Date
June 30, 2015

Project 2: Innovative approaches to gauge progression of Sturge-Weber syndrome

Administered By
Molecular Genetics and Microbiology
AwardedBy
University of California - San Francisco
Role
Principal Investigator
Start Date
September 30, 2009
End Date
September 29, 2014

Natural genetic variation regulating infarct volume

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
August 01, 2009
End Date
July 31, 2014

Peripheral endothelial and muscle cell pathology in cardiovascular disease

Administered By
Medicine, Cardiology
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
August 20, 2010
End Date
June 30, 2013

Studying Early-Stage Lesions in Mouse Models of Cerebral Cavernous Malformations

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
December 01, 2011
End Date
May 12, 2013

Genesis and Progression of Cerebral Cavernous Malformations

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
May 01, 2009
End Date
February 28, 2013

Genetic modifiers of dilated cardiomyopathy in adult Drosophila

Administered By
Medicine, Cardiology
AwardedBy
National Institutes of Health
Role
Significant Contributor
Start Date
August 01, 2007
End Date
June 30, 2012

Identification of genetics modifiers of heart disease

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
February 20, 2007
End Date
January 31, 2011

Genes That Regulate Progression of Kidney Disease and Its Cardiovascular Effects

Administered By
Medicine, Nephrology
AwardedBy
National Institutes of Health
Role
Consultant
Start Date
June 01, 2007
End Date
June 30, 2009

Investigation of the two-hit hypothesis for Cerebral Cavernous Malformations

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 01, 2008
End Date
May 10, 2009

Duke PREP: Minority Recruitment into Biomedical Sciences

Administered By
Biochemistry
AwardedBy
National Institutes of Health
Role
Advisor
Start Date
August 01, 2003
End Date
July 31, 2008

Identification of Modifier Genes in Heart Disease

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
June 20, 2005
End Date
June 19, 2008

Gene Discovery for Cerebral Cavernous Malformations

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
June 01, 2002
End Date
May 31, 2008

Identifying the CCM3 Gene: Understanding Familial Stroke

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
March 01, 2005
End Date
September 09, 2007

Modifier Genes in Heart Failure

Administered By
Medicine, Cardiology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
September 30, 2001
End Date
June 30, 2007

Mitochondrial DNA Stability and Mutagenesis

Administered By
Molecular Genetics and Microbiology
AwardedBy
Army Research Office
Role
Principal Investigator
Start Date
May 01, 2004
End Date
April 30, 2007

Genes that Regulate Target Organ Damage in Hypertension

Administered By
Medicine, Cardiology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
September 30, 2001
End Date
August 31, 2006

Genetics of Hereditary Hemorrhagic Telangiectasia

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
January 01, 1993
End Date
June 30, 2006

Investigating the function of the Mcgl Family

Administered By
Molecular Genetics and Microbiology
AwardedBy
Army Research Office
Role
Principal Investigator
Start Date
September 26, 2001
End Date
September 25, 2005

Characterization of the role of Din7 in mitochondrial DNA replication and genome integrity in the yeast Saccharomyces ce

Administered By
School of Medicine
AwardedBy
Army Research Office
Role
Principal Investigator
Start Date
May 01, 2002
End Date
April 30, 2004

A Mouse Model of Cerebral Cavernous Malformations

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
December 01, 2000
End Date
May 31, 2003

The Genetics Of Hereditary Hemorrhagic Telangiectasia

Administered By
School of Medicine
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
January 01, 1993
End Date
December 31, 1999

Diversity Of Tgf Beta Receptor Mutations In Hht Syndrome

Administered By
School of Medicine
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 01, 1995
End Date
August 31, 1998

Cloning The Hereditary Hemorrhagic Telangiectasia Gene

Administered By
School of Medicine
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
January 01, 1995
End Date
December 31, 1996

Cloning Of Hereditary Hemorrhagic Telangiectasia Gene

Administered By
School of Medicine
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
January 01, 1994
End Date
December 31, 1996
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Awards:

AAAS Fellows. American Association for the Advancement of Science, The.

Type
National
Awarded By
American Association for the Advancement of Science, The
Date
January 01, 2007

Publications:

RhoA Kinase Inhibition With Fasudil Versus Simvastatin in Murine Models of Cerebral Cavernous Malformations.

We sought to compare the effect of chronic treatment with commonly tolerated doses of Fasudil, a specific RhoA kinase (ROCK) inhibitor, and simvastatin (with pleiotropic effects including ROCK inhibition) on cerebral cavernous malformation (CCM) genesis and maturation in 2 models that recapitulate the human disease.Two heterozygous murine models, Ccm1+/-Msh2-/- and Ccm2+/-Trp53-/-, were treated from weaning to 4 to 5 months of age with Fasudil (100 mg/kg per day), simvastatin (40 mg/kg per day) or with placebo. Mouse brains were blindly assessed for CCM lesion burden, nonheme iron deposition (as a quantitative measure of chronic lesional hemorrhage), and ROCK activity.Fasudil, but not simvastatin, significantly decreased mature CCM lesion burden in Ccm1+/-Msh2-/- mice, and in meta-analysis of both models combined, when compared with mice receiving placebo. Fasudil and simvastatin both significantly decreased the integrated iron density per mature lesion area in Ccm1+/-Msh2-/- mice, and in both models combined, compared with mice given placebo. ROCK activity in mature lesions of Ccm1+/-Msh2-/- mice was similar with both treatments. Fasudil, but not simvastatin, improved survival in Ccm1+/-Msh2-/- mice. Fasudil and simvastatin treatment did not affect survival or lesion development significantly in Ccm2+/-Trp53-/- mice alone, and Fasudil benefit seemed limited to males.ROCK inhibitor Fasudil was more efficacious than simvastatin in improving survival and blunting the development of mature CCM lesions. Both drugs significantly decreased chronic hemorrhage in CCM lesions. These findings justify the development of ROCK inhibitors and the clinical testing of commonly used statin agents in CCM.

Authors
Shenkar, R; Shi, C; Austin, C; Moore, T; Lightle, R; Cao, Y; Zhang, L; Wu, M; Zeineddine, HA; Girard, R; McDonald, DA; Rorrer, A; Gallione, C; Pytel, P; Liao, JK; Marchuk, DA; Awad, IA
MLA Citation
Shenkar, R, Shi, C, Austin, C, Moore, T, Lightle, R, Cao, Y, Zhang, L, Wu, M, Zeineddine, HA, Girard, R, McDonald, DA, Rorrer, A, Gallione, C, Pytel, P, Liao, JK, Marchuk, DA, and Awad, IA. "RhoA Kinase Inhibition With Fasudil Versus Simvastatin in Murine Models of Cerebral Cavernous Malformations." Stroke 48.1 (January 2017): 187-194.
PMID
27879448
Source
epmc
Published In
Stroke
Volume
48
Issue
1
Publish Date
2017
Start Page
187
End Page
194
DOI
10.1161/strokeaha.116.015013

The pathobiology of vascular malformations: insights from human and model organism genetics.

Vascular malformations may arise in any of the vascular beds present in the human body. These lesions vary in location, type, and clinical severity of the phenotype. In recent years, the genetic basis of several vascular malformations has been elucidated. This review will consider how the identification of the genetic factors contributing to different vascular malformations, with subsequent functional studies in animal models, has provided a better understanding of these factors that maintain vascular integrity in vascular beds, as well as their role in the pathogenesis of vascular malformations. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Authors
Wetzel-Strong, SE; Detter, MR; Marchuk, DA
MLA Citation
Wetzel-Strong, SE, Detter, MR, and Marchuk, DA. "The pathobiology of vascular malformations: insights from human and model organism genetics." The Journal of pathology 241.2 (January 2017): 281-293.
PMID
27859310
Source
epmc
Published In
The Journal of Pathology
Volume
241
Issue
2
Publish Date
2017
Start Page
281
End Page
293
DOI
10.1002/path.4844

Micro-computed tomography in murine models of cerebral cavernous malformations as a paradigm for brain disease.

Cerebral cavernous malformations (CCMs) are hemorrhagic brain lesions, where murine models allow major mechanistic discoveries, ushering genetic manipulations and preclinical assessment of therapies. Histology for lesion counting and morphometry is essential yet tedious and time consuming. We herein describe the application and validations of X-ray micro-computed tomography (micro-CT), a non-destructive technique allowing three-dimensional CCM lesion count and volumetric measurements, in transgenic murine brains.We hereby describe a new contrast soaking technique not previously applied to murine models of CCM disease. Volumetric segmentation and image processing paradigm allowed for histologic correlations and quantitative validations not previously reported with the micro-CT technique in brain vascular disease.Twenty-two hyper-dense areas on micro-CT images, identified as CCM lesions, were matched by histology. The inter-rater reliability analysis showed strong consistency in the CCM lesion identification and staging (K=0.89, p<0.0001) between the two techniques. Micro-CT revealed a 29% greater CCM lesion detection efficiency, and 80% improved time efficiency.Serial integrated lesional area by histology showed a strong positive correlation with micro-CT estimated volume (r(2)=0.84, p<0.0001).Micro-CT allows high throughput assessment of lesion count and volume in pre-clinical murine models of CCM. This approach complements histology with improved accuracy and efficiency, and can be applied for lesion burden assessment in other brain diseases.

Authors
Girard, R; Zeineddine, HA; Orsbon, C; Tan, H; Moore, T; Hobson, N; Shenkar, R; Lightle, R; Shi, C; Fam, MD; Cao, Y; Shen, L; Neander, AI; Rorrer, A; Gallione, C; Tang, AT; Kahn, ML; Marchuk, DA; Luo, Z-X; Awad, IA
MLA Citation
Girard, R, Zeineddine, HA, Orsbon, C, Tan, H, Moore, T, Hobson, N, Shenkar, R, Lightle, R, Shi, C, Fam, MD, Cao, Y, Shen, L, Neander, AI, Rorrer, A, Gallione, C, Tang, AT, Kahn, ML, Marchuk, DA, Luo, Z-X, and Awad, IA. "Micro-computed tomography in murine models of cerebral cavernous malformations as a paradigm for brain disease." Journal of neuroscience methods 271 (September 2016): 14-24.
PMID
27345427
Source
epmc
Published In
Journal of Neuroscience Methods
Volume
271
Publish Date
2016
Start Page
14
End Page
24
DOI
10.1016/j.jneumeth.2016.06.021

Natural allelic variation of the IL-21 receptor modulates ischemic stroke infarct volume.

Risk for ischemic stroke has a strong genetic basis, but heritable factors also contribute to the extent of damage after a stroke has occurred. We previously identified a locus on distal mouse chromosome 7 that contributes over 50% of the variation in postischemic cerebral infarct volume observed between inbred strains. Here, we used ancestral haplotype analysis to fine-map this locus to 12 candidate genes. The gene encoding the IL-21 receptor (Il21r) showed a marked difference in strain-specific transcription levels and coding variants in neonatal and adult cortical tissue. Collateral vessel connections were moderately reduced in Il21r-deficient mice, and cerebral infarct volume increased 2.3-fold, suggesting that Il21r modulates both collateral vessel anatomy and innate neuroprotection. In brain slice explants, oxygen deprivation (OD) activated apoptotic pathways and increased neuronal cell death in IL-21 receptor-deficient (IL-21R-deficient) mice compared with control animals. We determined that the neuroprotective effects of IL-21R arose from signaling through JAK/STAT pathways and upregulation of caspase 3. Thus, natural genetic variation in murine Il21r influences neuronal cell viability after ischemia by modulating receptor function and downstream signal transduction. The identification of neuroprotective genes based on naturally occurring allelic variations has the potential to inform the development of drug targets for ischemic stroke treatment.

Authors
Lee, HK; Keum, S; Sheng, H; Warner, DS; Lo, DC; Marchuk, DA
MLA Citation
Lee, HK, Keum, S, Sheng, H, Warner, DS, Lo, DC, and Marchuk, DA. "Natural allelic variation of the IL-21 receptor modulates ischemic stroke infarct volume." The Journal of clinical investigation 126.8 (August 2016): 2827-2838.
PMID
27400126
Source
epmc
Published In
Journal of Clinical Investigation
Volume
126
Issue
8
Publish Date
2016
Start Page
2827
End Page
2838
DOI
10.1172/jci84491

B-Cell Depletion Reduces the Maturation of Cerebral Cavernous Malformations in Murine Models.

Cerebral cavernous malformations (CCMs) are relatively common vascular malformations, characterized by increased Rho kinase (ROCK) activity, vascular hyper-permeability and the presence of blood degradation products including non-heme iron. Previous studies revealed robust inflammatory cell infiltration, selective synthesis of IgG, in situ antigen driven B-cell clonal expansion, and deposition of immune complexes and complement proteins within CCM lesions. We aimed to evaluate the impact of suppressing the immune response on the formation and maturation of CCM lesions, as well as lesional iron deposition and ROCK activity. Two murine models of heterozygous Ccm3 (Pdcd10), which spontaneously develop CCM lesions with severe and milder phenotypes, were either untreated or received anti-mouse BR3 to deplete B cells. Brains from anti-mouse BR3-treated mice exhibited significantly fewer mature CCM lesions and smaller lesions compared to untreated mice. B cell depletion halted the progression of lesions into mature stage 2 lesions but did not prevent their genesis. Non-heme iron deposition and ROCK activity was decreased in lesions of B cell depleted mice. This represents the first report of the therapeutic benefit of B-cell depletion in the development and progression of CCMs, and provides a proof of principle that B cells play a critical role in CCM lesion genesis and maturation. These findings add biologics to the list of potential therapeutic agents for CCM disease. Future studies would characterize the putative antigenic trigger and further define the mechanism of immune response in the lesions.

Authors
Shi, C; Shenkar, R; Zeineddine, HA; Girard, R; Fam, MD; Austin, C; Moore, T; Lightle, R; Zhang, L; Wu, M; Cao, Y; Gunel, M; Louvi, A; Rorrer, A; Gallione, C; Marchuk, DA; Awad, IA
MLA Citation
Shi, C, Shenkar, R, Zeineddine, HA, Girard, R, Fam, MD, Austin, C, Moore, T, Lightle, R, Zhang, L, Wu, M, Cao, Y, Gunel, M, Louvi, A, Rorrer, A, Gallione, C, Marchuk, DA, and Awad, IA. "B-Cell Depletion Reduces the Maturation of Cerebral Cavernous Malformations in Murine Models." Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology 11.2 (June 2016): 369-377.
PMID
27086141
Source
epmc
Published In
Journal of NeuroImmune Pharmacology
Volume
11
Issue
2
Publish Date
2016
Start Page
369
End Page
377
DOI
10.1007/s11481-016-9670-0

Leveraging a Sturge-Weber Gene Discovery: An Agenda for Future Research

Authors
Comi, AM; Sahin, M; Hammill, A; Kaplan, EH; Juhász, C; North, P; Ball, KL; Levin, AV; Cohen, B; Morris, J; Lo, W; Roach, ES; Abreu, N; Acosta, M; Ball, KL; Berrocal, A; Bischoff, J; Brodie, J; Burkhart, C; Cohen, B; Comi, AM; Dymerska, G; Eckstein, D; Enriquez-Algeciras, M; Ewen, J; Fisher, B; Freedman, S; Germain-Lee, E; Geronemus, R; Gold, M; Gopal-Srivastava, R; Hammill, A; Hebert, A; Huang, L; Jampel, H; Juhász, C; Kaplan, EH; Kaseka, M; Kirkorian, Y; Kossoff, E; Levin, AV; Lin, D; Lo, W et al.
MLA Citation
Comi, AM, Sahin, M, Hammill, A, Kaplan, EH, Juhász, C, North, P, Ball, KL, Levin, AV, Cohen, B, Morris, J, Lo, W, Roach, ES, Abreu, N, Acosta, M, Ball, KL, Berrocal, A, Bischoff, J, Brodie, J, Burkhart, C, Cohen, B, Comi, AM, Dymerska, G, Eckstein, D, Enriquez-Algeciras, M, Ewen, J, Fisher, B, Freedman, S, Germain-Lee, E, Geronemus, R, Gold, M, Gopal-Srivastava, R, Hammill, A, Hebert, A, Huang, L, Jampel, H, Juhász, C, Kaplan, EH, Kaseka, M, Kirkorian, Y, Kossoff, E, Levin, AV, Lin, D, and Lo, W et al. "Leveraging a Sturge-Weber Gene Discovery: An Agenda for Future Research." Pediatric Neurology 58 (May 2016): 12-24.
Source
crossref
Published In
Pediatric Neurology
Volume
58
Publish Date
2016
Start Page
12
End Page
24
DOI
10.1016/j.pediatrneurol.2015.11.009

ADAM12: a genetic modifier of preclinical peripheral arterial disease.

In prior studies from multiple groups, outcomes following experimental peripheral arterial disease (PAD) differed considerably across inbred mouse strains. Similarly, in humans with PAD, disease outcomes differ, even when there are similarities in risk factors, disease anatomy, arteriosclerotic burden, and hemodynamic measures. Previously, we identified a locus on mouse chromosome 7, limb salvage-associated quantitative trait locus 1 (LSq-1), which was sufficient to modify outcomes following experimental PAD. We compared expression of genes within LSq-1 in Balb/c mice, which normally show poor outcomes following experimental PAD, with that in C57Bl/6 mice, which normally show favorable outcomes, and found that a disintegrin and metalloproteinase gene 12 (ADAM12) had the most differential expression. Augmentation of ADAM12 expression in vivo improved outcomes following experimental PAD in Balb/c mice, whereas knockdown of ADAM12 made outcomes worse in C57Bl/6 mice. In vitro, ADAM12 expression modulates endothelial cell proliferation, survival, and angiogenesis in ischemia, and this appeared to be dependent on tyrosine kinase with Ig-like and EGF-like domain 2 (Tie2) activation. ADAM12 is sufficient to modify PAD severity in mice, and this likely occurs through regulation of Tie2.

Authors
Dokun, AO; Chen, L; Okutsu, M; Farber, CR; Hazarika, S; Jones, WS; Craig, D; Marchuk, DA; Lye, RJ; Shah, SH; Annex, BH
MLA Citation
Dokun, AO, Chen, L, Okutsu, M, Farber, CR, Hazarika, S, Jones, WS, Craig, D, Marchuk, DA, Lye, RJ, Shah, SH, and Annex, BH. "ADAM12: a genetic modifier of preclinical peripheral arterial disease." American journal of physiology. Heart and circulatory physiology 309.5 (September 2015): H790-H803.
PMID
26163448
Source
epmc
Published In
American journal of physiology. Heart and circulatory physiology
Volume
309
Issue
5
Publish Date
2015
Start Page
H790
End Page
H803
DOI
10.1152/ajpheart.00803.2014

Exceptional aggressiveness of cerebral cavernous malformation disease associated with PDCD10 mutations.

The phenotypic manifestations of cerebral cavernous malformation disease caused by rare PDCD10 mutations have not been systematically examined, and a mechanistic link to Rho kinase-mediated hyperpermeability, a potential therapeutic target, has not been established.We analyzed PDCD10 small interfering RNA-treated endothelial cells for stress fibers, Rho kinase activity, and permeability. Rho kinase activity was assessed in cerebral cavernous malformation lesions. Brain permeability and cerebral cavernous malformation lesion burden were quantified, and clinical manifestations were assessed in prospectively enrolled subjects with PDCD10 mutations.We determined that PDCD10 protein suppresses endothelial stress fibers, Rho kinase activity, and permeability in vitro. Pdcd10 heterozygous mice have greater lesion burden than other Ccm genotypes. We demonstrated robust Rho kinase activity in murine and human cerebral cavernous malformation vasculature and increased brain vascular permeability in humans with PDCD10 mutation. Clinical phenotype is exceptionally aggressive compared with the more common KRIT1 and CCM2 familial and sporadic cerebral cavernous malformation, with greater lesion burden and more frequent hemorrhages earlier in life. We first report other phenotypic features, including scoliosis, cognitive disability, and skin lesions, unrelated to lesion burden or bleeding.These findings define a unique cerebral cavernous malformation disease with exceptional aggressiveness, and they inform preclinical therapeutic testing, clinical counseling, and the design of trials.Genet Med 17 3, 188-196.

Authors
Shenkar, R; Shi, C; Rebeiz, T; Stockton, RA; McDonald, DA; Mikati, AG; Zhang, L; Austin, C; Akers, AL; Gallione, CJ; Rorrer, A; Gunel, M; Min, W; Marcondes de Souza, J; Lee, C; Marchuk, DA; Awad, IA
MLA Citation
Shenkar, R, Shi, C, Rebeiz, T, Stockton, RA, McDonald, DA, Mikati, AG, Zhang, L, Austin, C, Akers, AL, Gallione, CJ, Rorrer, A, Gunel, M, Min, W, Marcondes de Souza, J, Lee, C, Marchuk, DA, and Awad, IA. "Exceptional aggressiveness of cerebral cavernous malformation disease associated with PDCD10 mutations." Genetics in medicine : official journal of the American College of Medical Genetics 17.3 (March 2015): 188-196.
PMID
25122144
Source
epmc
Published In
Genetics in Medicine
Volume
17
Issue
3
Publish Date
2015
Start Page
188
End Page
196
DOI
10.1038/gim.2014.97

Sturge-Weber Syndrome

© 2015 Elsevier Inc. All rights reserved.Sturge-Weber syndrome (SWS) is the association of the facial port-wine birthmark with malformed leptomeningeal blood vessels and abnormal venous eye vessels. Occurrence is sporadic and in both males and females and reported in all racial and ethnic backgrounds. The genetic cause accounting for SWS is a somatic mosaic mutation in the GNAQ gene encoding the Gαq protein. Testing for this somatic mosaic mutation may be useful in the future for differentiating a SWS diagnosis in these patients from other capillary malformation related syndromes, such as -megalencephaly-capillary malformation-polymicrogyria syndrome, which also have capillary malformations but are otherwise different in terms of prognosis and associated complications. With the discovery of the causative somatic mosaic mutation, current SWS research will likely result in new in vitro and animal models, potential novel treatment strategies, and new insights into the pathophysiology of this vascular malformation disorder.

Authors
Comi, AM; Marchuk, DA; Pevsner, J
MLA Citation
Comi, AM, Marchuk, DA, and Pevsner, J. "Sturge-Weber Syndrome." Rosenberg's Molecular and Genetic Basis of Neurological and Psychiatric Disease: Fifth Edition. November 13, 2014. 945-953.
Source
scopus
Publish Date
2014
Start Page
945
End Page
953
DOI
10.1016/B978-0-12-410529-4.00081-4

Lesions from patients with sporadic cerebral cavernous malformations harbor somatic mutations in the CCM genes: evidence for a common biochemical pathway for CCM pathogenesis.

Cerebral cavernous malformations (CCMs) are vascular lesions affecting the central nervous system. CCM occurs either sporadically or in an inherited, autosomal dominant manner. Constitutional (germline) mutations in any of three genes, KRIT1, CCM2 and PDCD10, can cause the inherited form. Analysis of CCM lesions from inherited cases revealed biallelic somatic mutations, indicating that CCM follows a Knudsonian two-hit mutation mechanism. It is still unknown, however, if the sporadic cases of CCM also follow this genetic mechanism. We extracted DNA from 11 surgically excised lesions from sporadic CCM patients, and sequenced the three CCM genes in each specimen using a next-generation sequencing approach. Four sporadic CCM lesion samples (36%) were found to contain novel somatic mutations. Three of the lesions contained a single somatic mutation, and one lesion contained two biallelic somatic mutations. Herein, we also describe evidence of somatic mosaicism in a patient presenting with over 130 CCM lesions localized to one hemisphere of the brain. Finally, in a lesion regrowth sample, we found that the regrown CCM lesion contained the same somatic mutation as the original lesion. Together, these data bolster the idea that all forms of CCM have a genetic underpinning of the two-hit mutation mechanism in the known CCM genes. Recent studies have found aberrant Rho kinase activation in inherited CCM pathogenesis, and we present evidence that this pathway is activated in sporadic CCM patients. These results suggest that all CCM patients, including those with the more common sporadic form, are potentially amenable to the same therapy.

Authors
McDonald, DA; Shi, C; Shenkar, R; Gallione, CJ; Akers, AL; Li, S; De Castro, N; Berg, MJ; Corcoran, DL; Awad, IA; Marchuk, DA
MLA Citation
McDonald, DA, Shi, C, Shenkar, R, Gallione, CJ, Akers, AL, Li, S, De Castro, N, Berg, MJ, Corcoran, DL, Awad, IA, and Marchuk, DA. "Lesions from patients with sporadic cerebral cavernous malformations harbor somatic mutations in the CCM genes: evidence for a common biochemical pathway for CCM pathogenesis." Human molecular genetics 23.16 (August 2014): 4357-4370.
PMID
24698976
Source
epmc
Published In
Human Molecular Genetics
Volume
23
Issue
16
Publish Date
2014
Start Page
4357
End Page
4370
DOI
10.1093/hmg/ddu153

Endoglin deficiency impairs stroke recovery.

Endoglin deficiency causes hereditary hemorrhagic telangiectasia-1 and impairs myocardial repair. Pulmonary arteriovenous malformations in patients with hereditary hemorrhagic telangiectasia-1 are associated with a high incidence of paradoxical embolism in the cerebral circulation and ischemic brain injury. We hypothesized that endoglin deficiency impairs stroke recovery.Eng heterozygous (Eng+/-) and wild-type mice underwent permanent distal middle cerebral artery occlusion (pMCAO). Pial collateral vessels were quantified before pMCAO. Infarct/atrophic volume, vascular density, and macrophages were quantified in various days after pMCAO, and behavioral function was assessed using corner and adhesive removal tests on days 3, 15, 30, and 60 after pMCAO. The association between ENG 207G>A polymorphism and brain arteriovenous malformation rupture and surgery outcome was analyzed using logistic regression analysis in 256 ruptured and 157 unruptured patients.After pMCAO, Eng+/- mice showed larger infarct/atrophic volumes at all time points (P<0.05) and showed worse behavior performance (P<0.05) at 15, 30, and 60 days when compared with wild-type mice. Eng+/- mice had fewer macrophages on day 3 (P=0.009) and more macrophages on day 60 (P=0.02) in the peri-infarct region. Although Eng+/- and wild-type mice had similar numbers of pial collateral vessels before pMCAO, Eng+/- mice had lower vascular density in the peri-infarct region (P=0.05) on day 60 after pMCAO. In humans, ENG 207A allele has been associated with worse outcomes after arteriovenous malformation rupture or surgery of patients with unruptured arteriovenous malformation.Endoglin deficiency impairs brain injury recovery. Reduced angiogenesis, impaired macrophage homing, and delayed inflammation resolution could be the underlying mechanism.

Authors
Shen, F; Degos, V; Chu, P-L; Han, Z; Westbroek, EM; Choi, E-J; Marchuk, D; Kim, H; Lawton, MT; Maze, M; Young, WL; Su, H
MLA Citation
Shen, F, Degos, V, Chu, P-L, Han, Z, Westbroek, EM, Choi, E-J, Marchuk, D, Kim, H, Lawton, MT, Maze, M, Young, WL, and Su, H. "Endoglin deficiency impairs stroke recovery." Stroke 45.7 (July 2014): 2101-2106.
PMID
24876084
Source
epmc
Published In
Stroke
Volume
45
Issue
7
Publish Date
2014
Start Page
2101
End Page
2106
DOI
10.1161/strokeaha.114.005115

Inhibition of the cardiomyocyte-specific troponin I-interacting kinase limits oxidative stress, injury, and adverse remodeling due to ischemic heart disease.

Ischemia–reperfusion injury is strongly associated with increased oxidative stress, mitochondrial dysfunction, and cell death. These processes are diminished in an animal model of ischemia–reperfusion by the genetic loss or pharmacological inhibition of troponin I–interacting kinase.

Authors
Abraham, DM; Marchuk, DA
MLA Citation
Abraham, DM, and Marchuk, DA. "Inhibition of the cardiomyocyte-specific troponin I-interacting kinase limits oxidative stress, injury, and adverse remodeling due to ischemic heart disease." Circulation research 114.6 (March 2014): 938-940.
PMID
24625723
Source
epmc
Published In
Circulation Research
Volume
114
Issue
6
Publish Date
2014
Start Page
938
End Page
940
DOI
10.1161/circresaha.113.303238

EndoU is a novel regulator of AICD during peripheral B cell selection.

Balanced transmembrane signals maintain a competent peripheral B cell pool limited in self-reactive B cells that may produce pathogenic autoantibodies. To identify molecules regulating peripheral B cell survival and tolerance to self-antigens (Ags), a gene modifier screen was performed with B cells from CD22-deficient C57BL/6 (CD22(-/-[B6])) mice that undergo activation-induced cell death (AICD) and fail to up-regulate c-Myc expression after B cell Ag receptor ligation. Likewise, lysozyme auto-Ag-specific B cells in Ig(Tg) hen egg lysozyme (HEL) transgenic mice inhabit the spleen but undergo AICD after auto-Ag encounter. This gene modifier screen identified EndoU, a single-stranded RNA-binding protein of ancient origin, as a major regulator of B cell survival in both models. EndoU gene disruption prevents AICD and normalizes c-Myc expression. These findings reveal that EndoU is a critical regulator of an unexpected and novel RNA-dependent pathway controlling peripheral B cell survival and Ag responsiveness that may contribute to peripheral B cell tolerance.

Authors
Poe, JC; Kountikov, EI; Lykken, JM; Natarajan, A; Marchuk, DA; Tedder, TF
MLA Citation
Poe, JC, Kountikov, EI, Lykken, JM, Natarajan, A, Marchuk, DA, and Tedder, TF. "EndoU is a novel regulator of AICD during peripheral B cell selection." J Exp Med 211.1 (January 13, 2014): 57-69.
PMID
24344237
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
211
Issue
1
Publish Date
2014
Start Page
57
End Page
69
DOI
10.1084/jem.20130648

A needle in a haystack: Sturge-Weber syndrome gene discovery

Authors
Comi, AM; Marchuk, DA; Pevsner, J
MLA Citation
Comi, AM, Marchuk, DA, and Pevsner, J. "A needle in a haystack: Sturge-Weber syndrome gene discovery." Pediatric Neurology 49.6 (December 1, 2013): 391-392. (Review)
Source
scopus
Published In
Pediatric Neurology
Volume
49
Issue
6
Publish Date
2013
Start Page
391
End Page
392
DOI
10.1016/j.pediatrneurol.2013.07.009

A needle in a haystack: Sturge-Weber syndrome gene discovery.

Authors
Comi, AM; Marchuk, DA; Pevsner, J
MLA Citation
Comi, AM, Marchuk, DA, and Pevsner, J. "A needle in a haystack: Sturge-Weber syndrome gene discovery." Pediatr Neurol 49.6 (December 2013): 391-392.
PMID
24075845
Source
pubmed
Published In
Pediatric Neurology
Volume
49
Issue
6
Publish Date
2013
Start Page
391
End Page
392
DOI
10.1016/j.pediatrneurol.2013.07.009

A novel genetic locus modulates infarct volume independently of the extent of collateral circulation.

In the mouse model of permanent, middle cerebral artery occlusion, infarct volume varies widely across inbred strains but generally is inversely correlated with collateral vessel number. However, we also observed certain mouse strains that share similar collateral vessel anatomy but exhibit significantly different infarct volume. To identify genetic factors determining infarct volume in a collateral vessel-independent manner, we performed quantitative trait locus analysis on a F2 cross between C57BL/6J and C3H/HeJ strains. We mapped four novel loci (Civq4 through Civq7) that modulate infarct volume. Civq4, on chromosome 8, is the strongest locus (logarithm of the odds 9.8) that contributes 21% of the phenotypic variance of infarct volume in the cross. The Civq4 and Civq6 loci represent transgressive B6 alleles that render animals susceptible to larger infarcts. Based on genomic sequence and microarray analyses, we propose candidate genes for the Civq4 locus. By selecting inbred strains with similar collateral vessel anatomy but that vary significantly in infarct volume, we have mapped four loci determining infarct volume in a mouse model of ischemic stroke. Two of the loci appear to modulate infarct volume through a collateral vessel-independent mechanism. Based on strain-specific sequence variants and differences in transcript levels, Msr1 and Mtmr7 appear to be strong candidate genes for Civq4. Identifying the underlying genetic factors of these loci will elucidate the genetic architecture response to cerebral ischemia, shed new light on disease mechanisms of ischemic stroke, and identify potential therapeutic targets for clinical applications.

Authors
Chu, P-L; Keum, S; Marchuk, DA
MLA Citation
Chu, P-L, Keum, S, and Marchuk, DA. "A novel genetic locus modulates infarct volume independently of the extent of collateral circulation." Physiol Genomics 45.17 (September 3, 2013): 751-763.
PMID
23800850
Source
pubmed
Published In
Physiological genomics
Volume
45
Issue
17
Publish Date
2013
Start Page
751
End Page
763
DOI
10.1152/physiolgenomics.00063.2013

Sturge-Weber syndrome and port-wine stains caused by somatic mutation in GNAQ.

BACKGROUND: The Sturge-Weber syndrome is a sporadic congenital neurocutaneous disorder characterized by a port-wine stain affecting the skin in the distribution of the ophthalmic branch of the trigeminal nerve, abnormal capillary venous vessels in the leptomeninges of the brain and choroid, glaucoma, seizures, stroke, and intellectual disability. It has been hypothesized that somatic mosaic mutations disrupting vascular development cause both the Sturge-Weber syndrome and port-wine stains, and the severity and extent of presentation are determined by the developmental time point at which the mutations occurred. To date, no such mutation has been identified. METHODS: We performed whole-genome sequencing of DNA from paired samples of visibly affected and normal tissue from 3 persons with the Sturge-Weber syndrome. We tested for the presence of a somatic mosaic mutation in 97 samples from 50 persons with the Sturge-Weber syndrome, a port-wine stain, or neither (controls), using amplicon sequencing and SNaPshot assays, and investigated the effects of the mutation on downstream signaling, using phosphorylation-specific antibodies for relevant effectors and a luciferase reporter assay. RESULTS: We identified a nonsynonymous single-nucleotide variant (c.548G→A, p.Arg183Gln) in GNAQ in samples of affected tissue from 88% of the participants (23 of 26) with the Sturge-Weber syndrome and from 92% of the participants (12 of 13) with apparently nonsyndromic port-wine stains, but not in any of the samples of affected tissue from 4 participants with an unrelated cerebrovascular malformation or in any of the samples from the 6 controls. The prevalence of the mutant allele in affected tissues ranged from 1.0 to 18.1%. Extracellular signal-regulated kinase activity was modestly increased during transgenic expression of mutant Gαq. CONCLUSIONS: The Sturge-Weber syndrome and port-wine stains are caused by a somatic activating mutation in GNAQ. This finding confirms a long-standing hypothesis. (Funded by the National Institutes of Health and Hunter's Dream for a Cure Foundation.).

Authors
Shirley, MD; Tang, H; Gallione, CJ; Baugher, JD; Frelin, LP; Cohen, B; North, PE; Marchuk, DA; Comi, AM; Pevsner, J
MLA Citation
Shirley, MD, Tang, H, Gallione, CJ, Baugher, JD, Frelin, LP, Cohen, B, North, PE, Marchuk, DA, Comi, AM, and Pevsner, J. "Sturge-Weber syndrome and port-wine stains caused by somatic mutation in GNAQ." N Engl J Med 368.21 (May 23, 2013): 1971-1979.
PMID
23656586
Source
pubmed
Published In
The New England journal of medicine
Volume
368
Issue
21
Publish Date
2013
Start Page
1971
End Page
1979
DOI
10.1056/NEJMoa1213507

Overexpression of TNNI3K, a cardiac-specific MAPKKK, promotes cardiac dysfunction.

Cardiac troponin I-interacting kinase (TNNI3K) is a cardiac-specific kinase whose biological function remains largely unknown. We have recently shown that TNNI3K expression greatly accelerates cardiac dysfunction in mouse models of cardiomyopathy, indicating an important role in modulating disease progression. To further investigate TNNI3K kinase activity in vivo, we have generated transgenic mice expressing both wild-type and kinase-dead versions of the human TNNI3K protein. Importantly, we show that the increased TNNI3K kinase activity induces mouse cardiac remodeling, and its kinase activity promotes accelerated disease progression in a left-ventricular pressure overload model of mouse cardiomyopathy. Using an in vitro kinase assay and proteomics analysis, we show that TNNI3K is a dual-function kinase with Tyr and Ser/Thr kinase activity. TNNI3K expression induces a series of cellular and molecular changes, including a reduction of sarcomere length and changes in titin isoform composition, which are indicative of cardiac remodeling. Using antisera to TNNI3K, we show that TNNI3K protein is located at the sarcomere Z disc. These combined data suggest that TNNI3K mediates cell signaling to modulate cardiac response to stress.

Authors
Tang, H; Xiao, K; Mao, L; Rockman, HA; Marchuk, DA
MLA Citation
Tang, H, Xiao, K, Mao, L, Rockman, HA, and Marchuk, DA. "Overexpression of TNNI3K, a cardiac-specific MAPKKK, promotes cardiac dysfunction." J Mol Cell Cardiol 54 (January 2013): 101-111.
PMID
23085512
Source
pubmed
Published In
Journal of Molecular and Cellular Cardiology
Volume
54
Publish Date
2013
Start Page
101
End Page
111
DOI
10.1016/j.yjmcc.2012.10.004

Overexpression of TNNI3K, a cardiac-specific MAPKKK, promotes cardiac dysfunction

Cardiac troponin I-interacting kinase (TNNI3K) is a cardiac-specific kinase whose biological function remains largely unknown. We have recently shown that TNNI3K expression greatly accelerates cardiac dysfunction in mouse models of cardiomyopathy, indicating an important role in modulating disease progression. To further investigate TNNI3K kinase activity in vivo, we have generated transgenic mice expressing both wild-type and kinase-dead versions of the human TNNI3K protein. Importantly, we show that the increased TNNI3K kinase activity induces mouse cardiac remodeling, and its kinase activity promotes accelerated disease progression in a left-ventricular pressure overload model of mouse cardiomyopathy. Using an in vitro kinase assay and proteomics analysis, we show that TNNI3K is a dual-function kinase with Tyr and Ser/Thr kinase activity. TNNI3K expression induces a series of cellular and molecular changes, including a reduction of sarcomere length and changes in titin isoform composition, which are indicative of cardiac remodeling. Using antisera to TNNI3K, we show that TNNI3K protein is located at the sarcomere Z disc. These combined data suggest that TNNI3K mediates cell signaling to modulate cardiac response to stress. © 2012 Elsevier Ltd.

Authors
Tang, H; Xiao, K; Mao, L; Rockman, HA; Marchuk, DA
MLA Citation
Tang, H, Xiao, K, Mao, L, Rockman, HA, and Marchuk, DA. "Overexpression of TNNI3K, a cardiac-specific MAPKKK, promotes cardiac dysfunction." Journal of Molecular and Cellular Cardiology 54.1 (2013): 101-111.
Source
scival
Published In
Journal of Molecular and Cellular Cardiology
Volume
54
Issue
1
Publish Date
2013
Start Page
101
End Page
111
DOI
10.1016/j.yjmcc.2012.10.004

Hereditary Hemorrhagic Telangiectasia (Osler-Weber-Rendu Syndrome)

Authors
Guttmacher, AE; Marchuk, DA; Trerotola, SO; Pyeritz, RE
MLA Citation
Guttmacher, AE, Marchuk, DA, Trerotola, SO, and Pyeritz, RE. "Hereditary Hemorrhagic Telangiectasia (Osler-Weber-Rendu Syndrome)." Emery and Rimoin's Principles and Practice of Medical Genetics (2013): 1-18.
Source
scival
Published In
Emery and Rimoin's Principles and Practice of Medical Genetics
Publish Date
2013
Start Page
1
End Page
18
DOI
10.1016/B978-0-12-383834-6.00055-0

Natural genetic variation of integrin alpha L (Itgal) modulates ischemic brain injury in stroke.

During ischemic stroke, occlusion of the cerebrovasculature causes neuronal cell death (infarction), but naturally occurring genetic factors modulating infarction have been difficult to identify in human populations. In a surgically induced mouse model of ischemic stroke, we have previously mapped Civq1 to distal chromosome 7 as a quantitative trait locus determining infarct volume. In this study, genome-wide association mapping using 32 inbred mouse strains and an additional linkage scan for infarct volume confirmed that the size of the infarct is determined by ancestral alleles of the causative gene(s). The genetically isolated Civq1 locus in reciprocal recombinant congenic mice refined the critical interval and demonstrated that infarct size is determined by both vascular (collateral vessel anatomy) and non-vascular (neuroprotection) effects. Through the use of interval-specific SNP haplotype analysis, we further refined the Civq1 locus and identified integrin alpha L (Itgal) as one of the causative genes for Civq1. Itgal is the only gene that exhibits both strain-specific amino acid substitutions and expression differences. Coding SNPs, a 5-bp insertion in exon 30b, and increased mRNA and protein expression of a splice variant of the gene (Itgal-003, ENSMUST00000120857), all segregate with infarct volume. Mice lacking Itgal show increased neuronal cell death in both ex vivo brain slice and in vivo focal cerebral ischemia. Our data demonstrate that sequence variation in Itgal modulates ischemic brain injury, and that infarct volume is determined by both vascular and non-vascular mechanisms.

Authors
Keum, S; Lee, HK; Chu, P-L; Kan, MJ; Huang, M-N; Gallione, CJ; Gunn, MD; Lo, DC; Marchuk, DA
MLA Citation
Keum, S, Lee, HK, Chu, P-L, Kan, MJ, Huang, M-N, Gallione, CJ, Gunn, MD, Lo, DC, and Marchuk, DA. "Natural genetic variation of integrin alpha L (Itgal) modulates ischemic brain injury in stroke." PLoS Genet 9.10 (2013): e1003807-.
PMID
24130503
Source
pubmed
Published In
PLoS genetics
Volume
9
Issue
10
Publish Date
2013
Start Page
e1003807
DOI
10.1371/journal.pgen.1003807

Skeletal muscle-specific genetic determinants contribute to the differential strain-dependent effects of hindlimb ischemia in mice.

Genetics plays an important role in determining peripheral arterial disease (PAD) pathology, which causes a spectrum of clinical disorders that range from clinically silent reductions in blood flow to limb-threatening ischemia. The cell-type specificity of PAD pathology, however, has received little attention. To determine whether strain-dependent differences in skeletal muscle cells might account for the differential responses to ischemia observed in C57BL/6 and BALB/c mice, endothelial and skeletal muscle cells were subjected to hypoxia and nutrient deprivation (HND) in vitro, to mimic ischemia. Muscle cells were more susceptible to HND than were endothelial cells. In vivo, C57BL/6 and BALB/c mice displayed strain-specific differences in myofiber responses after hindlimb ischemia, with significantly greater myofiber atrophy, greater apoptosis, and attenuated myogenic regulatory gene expression and stress-responsive signaling in BALB/c mice. Strain-specific deficits were recapitulated in vitro in primary muscle cells from both strains after HND. Muscle cells from BALB/c mice congenic for the C57BL/6 Lsq-1 quantitative trait locus were protected from HND-induced atrophy, and gene expression of vascular growth factors and their receptors was significantly greater in C57BL/6 primary muscle cells. Our results indicate that the previously identified specific genetic locus regulating strain-dependent collateral vessel density has a nonvascular or muscle cell-autonomous role involving both the myogenic program and traditional vascular growth factor receptor expression.

Authors
McClung, JM; McCord, TJ; Keum, S; Johnson, S; Annex, BH; Marchuk, DA; Kontos, CD
MLA Citation
McClung, JM, McCord, TJ, Keum, S, Johnson, S, Annex, BH, Marchuk, DA, and Kontos, CD. "Skeletal muscle-specific genetic determinants contribute to the differential strain-dependent effects of hindlimb ischemia in mice." Am J Pathol 180.5 (May 2012): 2156-2169.
PMID
22445571
Source
pubmed
Published In
The American journal of pathology
Volume
180
Issue
5
Publish Date
2012
Start Page
2156
End Page
2169
DOI
10.1016/j.ajpath.2012.01.032

Updates and future horizons on the understanding, diagnosis, and treatment of Sturge-Weber syndrome brain involvement.

AIM: To review recent developments in the understanding, diagnosis, and treatment of Sturge-Weber syndrome (SWS). METHOD: Members of the Brain Vascular Malformation Consortium Sturge-Weber Syndrome National Workgroup contributed their expertise to review the literature and present promising directions for research. RESULTS: The increasing number of reports dealing with SWS over the last decade reflects progress in the diagnosis and understanding of the neurological involvement. The proliferation of centers and advocacy groups to care for patients with SWS and to stimulate research has aided the development of new insights into the clinical manifestations and the pathophysiology of neurological progression, and the development of novel hypotheses to direct future research. Many key questions remain, but the tools and networks to answer them are being developed. INTERPRETATION: This review summarizes important new knowledge and presents new research directions that are likely to provide further insights, earlier diagnosis, improved treatments, and possibly, prevention of this syndrome.

Authors
Lo, W; Marchuk, DA; Ball, KL; Juhász, C; Jordan, LC; Ewen, JB; Comi, A; Brain Vascular Malformation Consortium National Sturge-Weber Syndrome Workgroup,
MLA Citation
Lo, W, Marchuk, DA, Ball, KL, Juhász, C, Jordan, LC, Ewen, JB, Comi, A, and Brain Vascular Malformation Consortium National Sturge-Weber Syndrome Workgroup, . "Updates and future horizons on the understanding, diagnosis, and treatment of Sturge-Weber syndrome brain involvement." Dev Med Child Neurol 54.3 (March 2012): 214-223. (Review)
PMID
22191476
Source
pubmed
Published In
Developmental Medicine & Child Neurology
Volume
54
Issue
3
Publish Date
2012
Start Page
214
End Page
223
DOI
10.1111/j.1469-8749.2011.04169.x

Fasudil decreases lesion burden in a murine model of cerebral cavernous malformation disease.

BACKGROUND AND PURPOSE: Cerebral cavernous malformations (CCMs) are characterized by grossly dilated capillaries, associated with vascular leak and hemorrhage, and occur in sporadic or inherited (autosomal-dominant) forms with mutations in 1 of 3 gene loci (CCM 1, 2 or 3). We previously reported that the CCM1 protein (KRIT1) localizes to endothelial cell-cell junctions and loss of KRIT1 leads to junctional instability associated with activation of RhoA and its effector Rho kinase. Although Rho kinase inhibition has been proposed as potential therapy for CCM, there has been no demonstration of a therapeutic effect on CCM lesion genesis in vivo. METHODS: Our recently generated a model of CCM1 disease (Ccm1(+/-)Msh2(-/-)) was treated with the Rho kinase inhibitor fasudil (100 mg/kg/day administered in drinking water from weaning to 5 months of age), or placebo, and blindly assessed CCM lesion burden by systematic survey of animals' brains. For comparison, we also assessed therapeutic effect in previously described Ccm2(+/-)Trp53(-/-) mice treated with the same dose and duration of fasudil and placebo. RESULTS: Fasudil-treated Ccm1(+/-)Msh2(-/-) mice had a significantly decreased prevalence of CCM lesions compared with placebo controls. Lesions in treated animals were smaller and less likely associated with hemorrhage, inflammation, and endothelial proliferation and exhibited decreased expression of Rho kinase activation biomarkers. A therapeutic effect was also documented in Ccm2(+/-)Trp53(-/-) mice. CONCLUSIONS: This represents the first report of therapeutic benefit of pharmacological therapy in development and progression of CCMs and indicates that Rho kinase activation is a critical step in CCM lesion genesis and maturation.

Authors
McDonald, DA; Shi, C; Shenkar, R; Stockton, RA; Liu, F; Ginsberg, MH; Marchuk, DA; Awad, IA
MLA Citation
McDonald, DA, Shi, C, Shenkar, R, Stockton, RA, Liu, F, Ginsberg, MH, Marchuk, DA, and Awad, IA. "Fasudil decreases lesion burden in a murine model of cerebral cavernous malformation disease." Stroke 43.2 (February 2012): 571-574.
PMID
22034008
Source
pubmed
Published In
Stroke
Volume
43
Issue
2
Publish Date
2012
Start Page
571
End Page
574
DOI
10.1161/STROKEAHA.111.625467

Dissection of a quantitative trait locus for PR interval duration identifies Tnni3k as a novel modulator of cardiac conduction.

Atrio-ventricular conduction disease is a common feature in Mendelian rhythm disorders associated with sudden cardiac death and is characterized by prolongation of the PR interval on the surface electrocardiogram (ECG). Prolongation of the PR interval is also a strong predictor of atrial fibrillation, the most prevalent sustained cardiac arrhythmia. Despite the significant genetic component in PR duration variability, the genes regulating PR interval duration remain largely elusive. We here aimed to dissect the quantitative trait locus (QTL) for PR interval duration that we previously mapped in murine F2 progeny of a sensitized 129P2 and FVBN/J cross. To determine the underlying gene responsible for this QTL, genome-wide transcriptional profiling was carried out on myocardial tissue from 109 F2 mice. Expression QTLs (eQTLs) were mapped and the PR interval QTL was inspected for the co-incidence of eQTLs. We further determined the correlation of each of these transcripts to the PR interval. Tnni3k was the only eQTL, mapping to the PR-QTL, with an established abundant cardiac-specific expression pattern and a significant correlation to PR interval duration. Genotype inspection in various inbred mouse strains revealed the presence of at least three independent haplotypes at the Tnni3k locus. Measurement of PR interval duration and Tnni3k mRNA expression levels in six inbred lines identified a positive correlation between the level of Tnni3k mRNA and PR interval duration. Furthermore, in DBA/2J mice overexpressing hTNNI3K, and in DBA.AKR.hrtfm2 congenic mice, which harbor the AKR/J "high-Tnni3k expression" haplotype in the DBA/2J genetic background, PR interval duration was prolonged as compared to DBA/2J wild-type mice ("low-Tnni3k expression" haplotype). Our data provide the first evidence for a role of Tnni3k in controlling the electrocardiographic PR interval indicating a function of Tnni3k in atrio-ventricular conduction.

Authors
Lodder, EM; Scicluna, BP; Milano, A; Sun, AY; Tang, H; Remme, CA; Moerland, PD; Tanck, MWT; Pitt, GS; Marchuk, DA; Bezzina, CR
MLA Citation
Lodder, EM, Scicluna, BP, Milano, A, Sun, AY, Tang, H, Remme, CA, Moerland, PD, Tanck, MWT, Pitt, GS, Marchuk, DA, and Bezzina, CR. "Dissection of a quantitative trait locus for PR interval duration identifies Tnni3k as a novel modulator of cardiac conduction." PLoS Genet 8.12 (2012): e1003113-.
PMID
23236294
Source
pubmed
Published In
PLoS genetics
Volume
8
Issue
12
Publish Date
2012
Start Page
e1003113
DOI
10.1371/journal.pgen.1003113

A founder mutation in the Ashkenazi Jewish population affecting messenger RNA splicing of the CCM2 gene causes cerebral cavernous malformations.

PURPOSE: Cerebral cavernous malformations can occur sporadically or are caused by mutations in one of three identified genes. Cerebral cavernous malformations often remain clinically silent until a mutation carrier suffers a stroke or seizure. Presymptomatic genetic testing has been valuable to follow and manage cerebral cavernous malformation mutation carriers. During routine diagnostic testing, we identified a two base pair change in seven unrelated people of Ashkenazi Jewish heritage. Because of the location of the variant beyond the invariant splice donor sequence, the change was reported as a variant of unknown significance. In this study, we determined whether this change was a disease-causing mutation and whether it represents a founder mutation in the Ashkenazi Jewish population. METHODS: Transcripts arising from the normal and mutant alleles were examined by reverse transcription-polymerase chain reaction from affected and unaffected Ashkenazi Jewish cerebral cavernous malformation family members. A synthetic splicing system using a chimeric exon was used to visualize the effects of the change on splice donor site utilization. RESULTS: The two base pair change in CCM2, c.30 + 5_6delinsTT, segregated with affected status in the study families. Reverse transcription-polymerase chain reaction revealed loss of the transcript allele that was in phase with the mutation. The two base pair change, when tested in an in vitro synthetic splicing system, altered splice donor site utilization. Resequencing of the genomic region proximal and distal to the CCM2 gene mutation revealed a common single-nucleotide polymorphism haplotype in affected individuals. CONCLUSIONS: The two base pair change in CCM2, c.30 + 5_6delinsTT, disrupted proper splice donor utilization leading to a degraded transcript. Single nucleotide polymorphism haplotype analysis demonstrated that this mutation was due to a founder in the Ashkenazi Jewish population. These data have the potential to simplify genetic testing for cerebral cavernous malformation in the Ashkenazi Jewish population.

Authors
Gallione, CJ; Solatycki, A; Awad, IA; Weber, JL; Marchuk, DA
MLA Citation
Gallione, CJ, Solatycki, A, Awad, IA, Weber, JL, and Marchuk, DA. "A founder mutation in the Ashkenazi Jewish population affecting messenger RNA splicing of the CCM2 gene causes cerebral cavernous malformations." Genet Med 13.7 (July 2011): 662-666.
PMID
21543988
Source
pubmed
Published In
Genetics in Medicine
Volume
13
Issue
7
Publish Date
2011
Start Page
662
End Page
666
DOI
10.1097/GIM.0b013e318211ff8b

A novel mouse model of cerebral cavernous malformations based on the two-hit mutation hypothesis recapitulates the human disease.

Cerebral cavernous malformations (CCMs) are vascular lesions of the central nervous system appearing as multicavernous, blood-filled capillaries, leading to headache, seizure and hemorrhagic stroke. CCM occurs either sporadically or as an autosomal dominant disorder caused by germline mutation of one of the three genes: CCM1/KRIT1, CCM2/MGC4607 and CCM3/PDCD10. Surgically resected human CCM lesions have provided molecular and immunohistochemical evidence for a two-hit (germline plus somatic) mutation mechanism. In contrast to the equivalent human genotype, mice heterozygous for a Ccm1- or Ccm2-null allele do not develop CCM lesions. Based on the two-hit hypothesis, we attempted to improve the penetrance of the model by crossing Ccm1 and Ccm2 heterozygotes into a mismatch repair-deficient Msh2(-/-) background. Ccm1(+/-)Msh2(-/-) mice exhibit CCM lesions with high penetrance as shown by magnetic resonance imaging and histology. Significantly, the CCM lesions range in size from early-stage, isolated caverns to large, multicavernous lesions. A subset of endothelial cells within the CCM lesions revealed somatic loss of CCM protein staining, supporting the two-hit mutation mechanism. The late-stage CCM lesions displayed many of the characteristics of human CCM lesions, including hemosiderin deposits, immune cell infiltration, increased endothelial cell proliferation and increased Rho-kinase activity. Some of these characteristics were also seen, but to a lesser extent, in early-stage lesions. Tight junctions were maintained between CCM lesion endothelial cells, but gaps were evident between endothelial cells and basement membrane was defective. In contrast, the Ccm2(+/-)Msh2(-/-) mice lacked cerebrovascular lesions. The CCM1 mouse model provides an in vivo tool to investigate CCM pathogenesis and new therapies.

Authors
McDonald, DA; Shenkar, R; Shi, C; Stockton, RA; Akers, AL; Kucherlapati, MH; Kucherlapati, R; Brainer, J; Ginsberg, MH; Awad, IA; Marchuk, DA
MLA Citation
McDonald, DA, Shenkar, R, Shi, C, Stockton, RA, Akers, AL, Kucherlapati, MH, Kucherlapati, R, Brainer, J, Ginsberg, MH, Awad, IA, and Marchuk, DA. "A novel mouse model of cerebral cavernous malformations based on the two-hit mutation hypothesis recapitulates the human disease." Hum Mol Genet 20.2 (January 15, 2011): 211-222.
PMID
20940147
Source
pubmed
Published In
Human Molecular Genetics
Volume
20
Issue
2
Publish Date
2011
Start Page
211
End Page
222
DOI
10.1093/hmg/ddq433

Two genes on A/J chromosome 18 are associated with susceptibility to Staphylococcus aureus infection by combined microarray and QTL analyses.

Although it has recently been shown that A/J mice are highly susceptible to Staphylococcus aureus sepsis as compared to C57BL/6J, the specific genes responsible for this differential phenotype are unknown. Using chromosome substitution strains (CSS), we found that loci on chromosomes 8, 11, and 18 influence susceptibility to S. aureus sepsis in A/J mice. We then used two candidate gene selection strategies to identify genes on these three chromosomes associated with S. aureus susceptibility, and targeted genes identified by both gene selection strategies. First, we used whole genome transcription profiling to identify 191 (56 on chr. 8, 100 on chr. 11, and 35 on chr. 18) genes on our three chromosomes of interest that are differentially expressed between S. aureus-infected A/J and C57BL/6J. Second, we identified two significant quantitative trait loci (QTL) for survival post-infection on chr. 18 using N(2) backcross mice (F(1) [C18A]xC57BL/6J). Ten genes on chr. 18 (March3, Cep120, Chmp1b, Dcp2, Dtwd2, Isoc1, Lman1, Spire1, Tnfaip8, and Seh1l) mapped to the two significant QTL regions and were also identified by the expression array selection strategy. Using real-time PCR, 6 of these 10 genes (Chmp1b, Dtwd2, Isoc1, Lman1, Tnfaip8, and Seh1l) showed significantly different expression levels between S. aureus-infected A/J and C57BL/6J. For two (Tnfaip8 and Seh1l) of these 6 genes, siRNA-mediated knockdown of gene expression in S. aureus-challenged RAW264.7 macrophages induced significant changes in the cytokine response (IL-1 beta and GM-CSF) compared to negative controls. These cytokine response changes were consistent with those seen in S. aureus-challenged peritoneal macrophages from CSS 18 mice (which contain A/J chromosome 18 but are otherwise C57BL/6J), but not C57BL/6J mice. These findings suggest that two genes, Tnfaip8 and Seh1l, may contribute to susceptibility to S. aureus in A/J mice, and represent promising candidates for human genetic susceptibility studies.

Authors
Ahn, S-H; Deshmukh, H; Johnson, N; Cowell, LG; Rude, TH; Scott, WK; Nelson, CL; Zaas, AK; Marchuk, DA; Keum, S; Lamlertthon, S; Sharma-Kuinkel, BK; Sempowski, GD; Fowler, VG
MLA Citation
Ahn, S-H, Deshmukh, H, Johnson, N, Cowell, LG, Rude, TH, Scott, WK, Nelson, CL, Zaas, AK, Marchuk, DA, Keum, S, Lamlertthon, S, Sharma-Kuinkel, BK, Sempowski, GD, and Fowler, VG. "Two genes on A/J chromosome 18 are associated with susceptibility to Staphylococcus aureus infection by combined microarray and QTL analyses. (Published online)" PLoS Pathog 6.9 (September 2, 2010): e1001088-.
Website
http://hdl.handle.net/10161/13319
PMID
20824097
Source
pubmed
Published In
PLoS pathogens
Volume
6
Issue
9
Publish Date
2010
Start Page
e1001088
DOI
10.1371/journal.ppat.1001088

Functional conservation of human Spastin in a Drosophila model of autosomal dominant-hereditary spastic paraplegia.

Mutations in spastin are the most frequent cause of the neurodegenerative disease autosomal dominant-hereditary spastic paraplegia (AD-HSP). Drosophila melanogaster lacking spastin exhibit striking behavioral similarities to human patients suffering from AD-HSP, suggesting conservation of Spastin function between the species. Consistent with this, we show that exogenous expression of wild-type Drosophila or human spastin rescues behavioral and cellular defects in spastin null flies equivalently. This enabled us to generate genetically representative models of AD-HSP, which arises from dominant mutations in spastin rather than a complete loss of the gene. Flies co-expressing one copy of wild-type human spastin and one encoding the K388R catalytic domain mutation in the fly spastin null background, exhibit aberrant distal synapse morphology and microtubule distribution, similar to but less severe than spastin nulls. R388 or a separate nonsense mutation act dominantly and are furthermore sufficient to confer partial rescue, supporting in vitro evidence for additional, non-catalytic Spastin functions. Using this model, we tested the observation from human pedigrees that S44L and P45Q are trans-acting modifiers of mutations affecting the Spastin catalytic domain. As in humans, both L44 and Q45 are largely silent when heterozygous, but exacerbate mutant phenotypes when expressed in trans with R388. These transgenic 'AD-HSP' flies therefore provide a powerful and tractable model to enhance our understanding of the cellular and behavioral consequences of human spastin mutations and test hypotheses directly relevant to the human disease.

Authors
Du, F; Ozdowski, EF; Kotowski, IK; Marchuk, DA; Sherwood, NT
MLA Citation
Du, F, Ozdowski, EF, Kotowski, IK, Marchuk, DA, and Sherwood, NT. "Functional conservation of human Spastin in a Drosophila model of autosomal dominant-hereditary spastic paraplegia." Hum Mol Genet 19.10 (May 15, 2010): 1883-1896.
PMID
20154342
Source
pubmed
Published In
Human Molecular Genetics
Volume
19
Issue
10
Publish Date
2010
Start Page
1883
End Page
1896
DOI
10.1093/hmg/ddq064

Overlapping spectra of SMAD4 mutations in juvenile polyposis (JP) and JP-HHT syndrome.

Juvenile polyposis (JP) and hereditary hemorrhagic telangiectasia (HHT) are clinically distinct diseases caused by mutations in SMAD4 and BMPR1A (for JP) and endoglin and ALK1 (for HHT). Recently, a combined syndrome of JP-HHT was described that is also caused by mutations in SMAD4. Although both JP and JP-HHT are caused by SMAD4 mutations, a possible genotype:phenotype correlation was noted as all of the SMAD4 mutations in the JP-HHT patients were clustered in the COOH-terminal MH2 domain of the protein. If valid, this correlation would provide a molecular explanation for the phenotypic differences, as well as a pre-symptomatic diagnostic test to distinguish patients at risk for the overlapping but different clinical features of the disorders. In this study, we collected 19 new JP-HHT patients from which we identified 15 additional SMAD4 mutations. We also reviewed the literature for other reports of JP patients with HHT symptoms with confirmed SMAD4 mutations. Our combined results show that although the SMAD4 mutations in JP-HHT patients do show a tendency to cluster in the MH2 domain, mutations in other parts of the gene also cause the combined syndrome. Thus, any mutation in SMAD4 can cause JP-HHT. Any JP patient with a SMAD4 mutation is, therefore, at risk for the visceral manifestations of HHT and any HHT patient with SMAD4 mutation is at risk for early onset gastrointestinal cancer. In conclusion, a patient who tests positive for any SMAD4 mutation must be considered at risk for the combined syndrome of JP-HHT and monitored accordingly.

Authors
Gallione, C; Aylsworth, AS; Beis, J; Berk, T; Bernhardt, B; Clark, RD; Clericuzio, C; Danesino, C; Drautz, J; Fahl, J; Fan, Z; Faughnan, ME; Ganguly, A; Garvie, J; Henderson, K; Kini, U; Leedom, T; Ludman, M; Lux, A; Maisenbacher, M; Mazzucco, S; Olivieri, C; Ploos van Amstel, JK; Prigoda-Lee, N; Pyeritz, RE; Reardon, W; Vandezande, K; Waldman, JD; White, RI; Williams, CA; Marchuk, DA
MLA Citation
Gallione, C, Aylsworth, AS, Beis, J, Berk, T, Bernhardt, B, Clark, RD, Clericuzio, C, Danesino, C, Drautz, J, Fahl, J, Fan, Z, Faughnan, ME, Ganguly, A, Garvie, J, Henderson, K, Kini, U, Leedom, T, Ludman, M, Lux, A, Maisenbacher, M, Mazzucco, S, Olivieri, C, Ploos van Amstel, JK, Prigoda-Lee, N, Pyeritz, RE, Reardon, W, Vandezande, K, Waldman, JD, White, RI, Williams, CA, and Marchuk, DA. "Overlapping spectra of SMAD4 mutations in juvenile polyposis (JP) and JP-HHT syndrome." Am J Med Genet A 152A.2 (February 2010): 333-339.
PMID
20101697
Source
pubmed
Published In
American Journal of Medical Genetics Part A
Volume
152A
Issue
2
Publish Date
2010
Start Page
333
End Page
339
DOI
10.1002/ajmg.a.33206

A locus mapping to mouse chromosome 7 determines infarct volume in a mouse model of ischemic stroke.

BACKGROUND: In a mouse model of focal cerebral ischemia, infarct volume is highly variable and strain dependent, but the natural genetic determinants responsible for this difference remain unknown. To identify genetic determinants regulating ischemic neuronal damage and to dissect apart the role of individual genes and physiological mechanisms in infarction in mice, we performed quantitative trait locus analysis of surgically induced cerebral infarct volume. METHODS AND RESULTS: After permanent occlusion of the distal middle cerebral artery, infarct volume was determined for 16 inbred strains of mice, chromosome substitution strains, and for 2 intercross cohorts, F2 (B6xBALB/c) and F2 (B6xSWR/J). Genome-wide linkage analysis was performed for infarct volume as a quantitative trait. Infarct volume varied up to 30-fold between strains, with heritability estimated at 0.88. Overall, 3 quantitative trait locus were identified that modulate infarct volume, with a major locus (Civq1) on chromosome 7 accounting for >50% of the variation, with a combined LOD score of 21.7. Interval-specific single nucleotide polymorphism haplotype analysis for Civq1 results in 12 candidate genes. CONCLUSIONS: The extent of ischemic tissue damage after distal middle cerebral artery occlusion in inbred strains of mice is modulated by genetic variation mapping to at least 3 different loci. A single locus on chromosome 7 determines the majority of the observed variation in the trait. This locus seems to be identical to LSq1, a locus conferring limb salvage and reperfusion in a mouse model of hindlimb ischemia. The identification of the genes underlying these loci may uncover novel genetic and physiological pathways that modulate cerebral infarction and provide new targets for therapeutic intervention in ischemic stroke, and possibly other ischemic diseases.

Authors
Keum, S; Marchuk, DA
MLA Citation
Keum, S, and Marchuk, DA. "A locus mapping to mouse chromosome 7 determines infarct volume in a mouse model of ischemic stroke." Circ Cardiovasc Genet 2.6 (December 2009): 591-598.
PMID
20031639
Source
pubmed
Published In
Circulation: Cardiovascular Genetics
Volume
2
Issue
6
Publish Date
2009
Start Page
591
End Page
598
DOI
10.1161/CIRCGENETICS.109.883231

Tnni3k modifies disease progression in murine models of cardiomyopathy.

The Calsequestrin (Csq) transgenic mouse model of cardiomyopathy exhibits wide variation in phenotypic progression dependent on genetic background. Seven heart failure modifier (Hrtfm) loci modify disease progression and outcome. Here we report Tnni3k (cardiac Troponin I-interacting kinase) as the gene underlying Hrtfm2. Strains with the more susceptible phenotype exhibit high transcript levels while less susceptible strains show dramatically reduced transcript levels. This decrease is caused by an intronic SNP in low-transcript strains that activates a cryptic splice site leading to a frameshifted transcript, followed by nonsense-mediated decay of message and an absence of detectable protein. A transgenic animal overexpressing human TNNI3K alone exhibits no cardiac phenotype. However, TNNI3K/Csq double transgenics display severely impaired systolic function and reduced survival, indicating that TNNI3K expression modifies disease progression. TNNI3K expression also accelerates disease progression in a pressure-overload model of heart failure. These combined data demonstrate that Tnni3k plays a critical role in the modulation of different forms of heart disease, and this protein may provide a novel target for therapeutic intervention.

Authors
Wheeler, FC; Tang, H; Marks, OA; Hadnott, TN; Chu, P-L; Mao, L; Rockman, HA; Marchuk, DA
MLA Citation
Wheeler, FC, Tang, H, Marks, OA, Hadnott, TN, Chu, P-L, Mao, L, Rockman, HA, and Marchuk, DA. "Tnni3k modifies disease progression in murine models of cardiomyopathy." PLoS Genet 5.9 (September 2009): e1000647-.
PMID
19763165
Source
pubmed
Published In
PLoS genetics
Volume
5
Issue
9
Publish Date
2009
Start Page
e1000647
DOI
10.1371/journal.pgen.1000647

An N-ethyl-N-nitrosourea mutagenesis recessive screen identifies two candidate regions for murine cardiomyopathy that map to chromosomes 1 and 15.

N-ethyl-N-nitrosourea (ENU) mutagenesis screens have been successful for identifying genes that affect important biological processes and diseases. However, for heart-related phenotypes, these screens have been employed exclusively for developmental phenotypes, and to date no adult cardiomyopathy-causing genes have been discovered through a mutagenesis screen. To identify novel disease-causing and disease-modifying genes for cardiomyopathy, we performed an ENU recessive mutagenesis screen in adult mice. Using noninvasive echocardiography to screen for abnormalities in cardiac function, we identified a heritable cardiomyopathic phenotype in two families. To identify the chromosomal regions where the mutations are localized, we used a single nucleotide polymorphism (SNP) panel for genetic mapping of mouse mutations. This panel provided whole-genome linkage information and identified the mutagenized candidate regions at the proximal end of chromosome 1 (family EN1), and at the distal end of chromosome 15 (family EN25). We have identified 94 affected mice in family EN1 and have narrowed the candidate interval to 1 Mb. We have identified 20 affected mice in family EN25 and have narrowed the candidate interval to 12 Mb. The identification of the genes responsible for the observed phenotype in these families will be strong candidates for disease-causing or disease-modifying genes in patients with heart failure.

Authors
Fernandez, L; Marchuk, DA; Moran, JL; Beier, DR; Rockman, HA
MLA Citation
Fernandez, L, Marchuk, DA, Moran, JL, Beier, DR, and Rockman, HA. "An N-ethyl-N-nitrosourea mutagenesis recessive screen identifies two candidate regions for murine cardiomyopathy that map to chromosomes 1 and 15." Mamm Genome 20.5 (May 2009): 296-304.
PMID
19387734
Source
pubmed
Published In
Mammalian Genome
Volume
20
Issue
5
Publish Date
2009
Start Page
296
End Page
304
DOI
10.1007/s00335-009-9184-7

Biallelic somatic and germline mutations in cerebral cavernous malformations (CCMs): evidence for a two-hit mechanism of CCM pathogenesis.

Cerebral cavernous malformations (CCMs) are vascular anomalies of the central nervous system, comprising dilated blood-filled capillaries lacking structural support. The lesions are prone to rupture, resulting in seizures or hemorrhagic stroke. CCM can occur sporadically, manifesting as solitary lesions, but also in families, where multiple lesions generally occur. Familial cases follow autosomal-dominant inheritance due to mutations in one of three genes, CCM1/KRIT1, CCM2/malcavernin or CCM3/PDCD10. The difference in lesion burden between familial and sporadic CCM, combined with limited molecular data, suggests that CCM pathogenesis may follow a two-hit molecular mechanism, similar to that seen for tumor suppressor genes. In this study, we investigate the two-hit hypothesis for CCM pathogenesis. Through repeated cycles of amplification, subcloning and sequencing of multiple clones per amplicon, we identify somatic mutations that are otherwise invisible by direct sequencing of the bulk amplicon. Biallelic germline and somatic mutations were identified in CCM lesions from all three forms of inherited CCMs. The somatic mutations are found only in a subset of the endothelial cells lining the cavernous vessels and not in interstitial lesion cells. These data suggest that CCM lesion genesis requires complete loss of function for one of the CCM genes. Although widely expressed in the different cell types of the brain, these data also suggest a unique role for the CCM proteins in endothelial cell biology.

Authors
Akers, AL; Johnson, E; Steinberg, GK; Zabramski, JM; Marchuk, DA
MLA Citation
Akers, AL, Johnson, E, Steinberg, GK, Zabramski, JM, and Marchuk, DA. "Biallelic somatic and germline mutations in cerebral cavernous malformations (CCMs): evidence for a two-hit mechanism of CCM pathogenesis." Hum Mol Genet 18.5 (March 1, 2009): 919-930.
PMID
19088123
Source
pubmed
Published In
Human Molecular Genetics
Volume
18
Issue
5
Publish Date
2009
Start Page
919
End Page
930
DOI
10.1093/hmg/ddn430

The cerebral cavernous malformation signaling pathway promotes vascular integrity via Rho GTPases.

Cerebral cavernous malformation (CCM) is a common vascular dysplasia that affects both systemic and central nervous system blood vessels. Loss of function mutations in the CCM2 gene cause CCM. Here we show that targeted disruption of Ccm2 in mice results in failed lumen formation and early embryonic death through an endothelial cell autonomous mechanism. We show that CCM2 regulates endothelial cytoskeletal architecture, cell-to-cell interactions and lumen formation. Heterozygosity at Ccm2, a genotype equivalent to that in human CCM, results in impaired endothelial barrier function. On the basis of our biochemical studies indicating that loss of CCM2 results in activation of RHOA GTPase, we rescued the cellular phenotype and barrier function in heterozygous mice with simvastatin, a drug known to inhibit Rho GTPases. These data offer the prospect for pharmacological treatment of a human vascular dysplasia with a widely available and safe drug.

Authors
Whitehead, KJ; Chan, AC; Navankasattusas, S; Koh, W; London, NR; Ling, J; Mayo, AH; Drakos, SG; Jones, CA; Zhu, W; Marchuk, DA; Davis, GE; Li, DY
MLA Citation
Whitehead, KJ, Chan, AC, Navankasattusas, S, Koh, W, London, NR, Ling, J, Mayo, AH, Drakos, SG, Jones, CA, Zhu, W, Marchuk, DA, Davis, GE, and Li, DY. "The cerebral cavernous malformation signaling pathway promotes vascular integrity via Rho GTPases." Nat Med 15.2 (February 2009): 177-184.
PMID
19151728
Source
pubmed
Published In
Nature Medicine
Volume
15
Issue
2
Publish Date
2009
Start Page
177
End Page
184
DOI
10.1038/nm.1911

Erratum: The cerebral cavernous malformation signaling pathway promotes vascular integrity via Rho GTPases (Nature Medicine (2009) 15 (177-184))

Authors
Whitehead, KJ; Chan, AC; Navankasattusas, S; Koh, W; London, NR; Ling, J; Mayo, AH; Drakos, SG; Jones, CA; Zhu, W; Marchuk, DA; Davis, GE; Li, DY
MLA Citation
Whitehead, KJ, Chan, AC, Navankasattusas, S, Koh, W, London, NR, Ling, J, Mayo, AH, Drakos, SG, Jones, CA, Zhu, W, Marchuk, DA, Davis, GE, and Li, DY. "Erratum: The cerebral cavernous malformation signaling pathway promotes vascular integrity via Rho GTPases (Nature Medicine (2009) 15 (177-184))." Nature Medicine 15.4 (2009): 462--.
Source
scival
Published In
Nature Medicine
Volume
15
Issue
4
Publish Date
2009
Start Page
462-
DOI
10.1038/nm0409-462c

Increased tissue perfusion promotes capillary dysplasia in the ALK1-deficient mouse brain following VEGF stimulation.

Loss-of-function activin receptor-like kinase 1 gene mutation (ALK1+/-) is associated with brain arteriovenous malformations (AVM) in hereditary hemorrhagic telangiectasia type 2. Other determinants of the lesional phenotype are unknown. In the present study, we investigated the influence of high vascular flow rates on ALK1+/- mice by manipulating cerebral blood flow (CBF) using vasodilators. Adult male ALK1+/- mice underwent adeno-associated viral-mediated vascular endothelial growth factor (AAVVEGF) or lacZ (AAVlacZ as a control) gene transfer into the brain. Two weeks after vector injection, hydralazine or nicardipine was infused intraventricularly for another 14 days. CBF was measured to evaluate relative tissue perfusion. We analyzed the number and morphology of capillaries. Results demonstrated that hydralazine or nicardipine infusion increased focal brain perfusion in all mice. It was noted that focal CBF increased most in AAVVEGF-injected ALK1+/- mice following hydralazine or nicardipine infusion (145+/-23% or 150+/-11%; P<0.05). There were more detectable dilated and dysplastic capillaries (2.4+/-0.3 or 2.0+/-0.4 dysplasia index; P<0.01) in the brains of ALK1+/- mice treated with AAVVEGF and hydralazine or nicardipine compared with the mice treated with them individually. We concluded that increased focal tissue perfusion and angiogenic factor VEGF stimulation could have a synergistic effect to promote capillary dysplasia in a genetic deficit animal model, which may have relevance to further studies of AVMs.

Authors
Hao, Q; Su, H; Marchuk, DA; Rola, R; Wang, Y; Liu, W; Young, WL; Yang, G-Y
MLA Citation
Hao, Q, Su, H, Marchuk, DA, Rola, R, Wang, Y, Liu, W, Young, WL, and Yang, G-Y. "Increased tissue perfusion promotes capillary dysplasia in the ALK1-deficient mouse brain following VEGF stimulation." Am J Physiol Heart Circ Physiol 295.6 (December 2008): H2250-H2256.
PMID
18835925
Source
pubmed
Published In
American journal of physiology. Heart and circulatory physiology
Volume
295
Issue
6
Publish Date
2008
Start Page
H2250
End Page
H2256
DOI
10.1152/ajpheart.00083.2008

Advanced magnetic resonance imaging of cerebral cavernous malformations: part II. Imaging of lesions in murine models.

OBJECTIVE: We sought to assess the appearance of cerebral cavernous malformations (CCM) on magnetic resonance imaging (MRI) scans in murine Ccm1 and Ccm2 gene knockout models and to develop a technique of lesion localization for correlative pathobiological studies METHODS: Brains from 18 CCM mutant mice (Ccm1 Trp53 and Ccm2 Trp53) and 28 control animals were imaged by gradient recalled echo (T2*)-weighted MRI scans at 4.7- and 14.1-T in vivo and/or ex vivo. After MRI scanning, the brains were removed and stained with hematoxylin and eosin, and cells were laser-microdissected for molecular biological studies. RESULTS: T2*-weighted MRI scans of brains in vivo and ex vivo revealed lesions similar to human CCMs in mutant mice, but not in control animals. Stereotactic localization and hematoxylin and eosin staining of correlative tissue sections confirmed lesion histology and revealed other areas of dilated capillaries in the same brains. Some lesions were identified by MRI scans at 14.1-T, but not at 4.7-T. Polymerase chain reaction amplification from Ccm1 and beta-actin genes was demonstrated from nucleic acids extracted from laser microdissected lesional and perilesional cells. CONCLUSION: The high-field MRI techniques offer new opportunities for further investigation of disease pathogenesis in vivo, and the localization, staging, and histobiological dissection of lesions, including the presumed earliest stages of CCM lesion development.

Authors
Shenkar, R; Venkatasubramanian, PN; Wyrwicz, AM; Zhao, J-C; Shi, C; Akers, A; Marchuk, DA; Awad, IA
MLA Citation
Shenkar, R, Venkatasubramanian, PN, Wyrwicz, AM, Zhao, J-C, Shi, C, Akers, A, Marchuk, DA, and Awad, IA. "Advanced magnetic resonance imaging of cerebral cavernous malformations: part II. Imaging of lesions in murine models." Neurosurgery 63.4 (October 2008): 790-797.
PMID
18981891
Source
pubmed
Published In
Neurosurgery
Volume
63
Issue
4
Publish Date
2008
Start Page
790
End Page
797
DOI
10.1227/01.NEU.0000315862.24920.49

ZPLD1 gene is disrupted in a patient with balanced translocation that exhibits cerebral cavernous malformations.

The past few years have seen rapid advances in our understanding of the genetics and molecular biology of cerebral cavernous malformations (CCM) with the identification of the CCM1, CCM2, and CCM3 genes. Recently, we have recruited a patient with an X/3 balanced translocation that exhibits CCM. By fluorescent in situ hybridization analysis, sequence analysis tools and database mining procedures, we refined the critical region to an interval of 200-kb and identified the interrupted ZPLD1 gene. We detected that the mRNA expression level of ZPLD1 gene is consistently decreased 2.5-fold versus control (P=0.0006) with allelic loss of gene expression suggesting that this protein may be part of the complex signaling pathway implicated in CCM formation.

Authors
Gianfrancesco, F; Esposito, T; Penco, S; Maglione, V; Liquori, CL; Patrosso, MC; Zuffardi, O; Ciccodicola, A; Marchuk, DA; Squitieri, F
MLA Citation
Gianfrancesco, F, Esposito, T, Penco, S, Maglione, V, Liquori, CL, Patrosso, MC, Zuffardi, O, Ciccodicola, A, Marchuk, DA, and Squitieri, F. "ZPLD1 gene is disrupted in a patient with balanced translocation that exhibits cerebral cavernous malformations." Neuroscience 155.2 (August 13, 2008): 345-349.
PMID
18632209
Source
pubmed
Published In
Neuroscience
Volume
155
Issue
2
Publish Date
2008
Start Page
345
End Page
349
DOI
10.1016/j.neuroscience.2008.05.030

A quantitative trait locus (LSq-1) on mouse chromosome 7 is linked to the absence of tissue loss after surgical hindlimb ischemia.

BACKGROUND: Peripheral arterial disease (PAD) caused by occlusive atherosclerosis of the lower extremity has 2 major clinical manifestations. Critical limb ischemia is characterized by rest pain and/or tissue loss and has a > or = 40% risk of death and major amputation. Intermittent claudication causes pain on walking, has no tissue loss, and has amputation plus mortality rates of 2% to 4% per year. Progression from claudication to limb ischemia is infrequent. Risk factors in most PAD patients overlap. Thus, we hypothesized that genetic variations may be linked to presence or absence of tissue loss in PAD. METHODS AND RESULTS: Hindlimb ischemia (murine model of PAD) was induced in C57BL/6, BALB/c, C57BL/6 x BALB/c (F1), F1 x BALB/c (N2), A/J, and C57BL/6J-Chr7(A/J)/NaJ chromosome substitution strains. Mice were monitored for perfusion recovery and tissue necrosis. Genome-wide scanning with polymorphic markers across the 19 murine autosomes was performed on the N2 mice. Greater tissue loss and poorer perfusion recovery occurred in BALB/c than in the C57BL/6 strain. Analysis of 105 N2 progeny identified a single quantitative trait locus on chromosome 7 that exhibited significant linkage to both tissue necrosis and extent of perfusion recovery. Using the appropriate chromosome substitution strain, we demonstrate that C57BL/6-derived chromosome 7 is required for tissue preservation. CONCLUSIONS: We have identified a quantitative trait locus on murine chromosome 7 (LSq-1) that is associated with the absence of tissue loss in a preclinical model of PAD and may be useful in identifying gene(s) that influence PAD in humans.

Authors
Dokun, AO; Keum, S; Hazarika, S; Li, Y; Lamonte, GM; Wheeler, F; Marchuk, DA; Annex, BH
MLA Citation
Dokun, AO, Keum, S, Hazarika, S, Li, Y, Lamonte, GM, Wheeler, F, Marchuk, DA, and Annex, BH. "A quantitative trait locus (LSq-1) on mouse chromosome 7 is linked to the absence of tissue loss after surgical hindlimb ischemia." Circulation 117.9 (March 4, 2008): 1207-1215.
PMID
18285563
Source
pubmed
Published In
Circulation
Volume
117
Issue
9
Publish Date
2008
Start Page
1207
End Page
1215
DOI
10.1161/CIRCULATIONAHA.107.736447

Different spectra of genomic deletions within the CCM genes between Italian and American CCM patient cohorts.

Cerebral cavernous malformations (CCMs) are vascular abnormalities of the brain that can result in hemorrhagic stroke and seizures. Familial forms of CCM are inherited in an autosomal-dominant fashion, and three CCM genes have been identified. We recently determined that large genomic deletions in the CCM2 gene represent 22% of mutations in a large CCM cohort from the USA. In particular, a 77.6 kb deletion spanning CCM2 exons 2-10 displays an identical recombination event in eight CCM probands/families and appears to be common in the US population. In the current study, we report the identification of six additional probands/families from the USA with this same large deletion. Haplotype analysis strongly suggests that this common deletion derives from an ancestral founder. We also examined an Italian CCM cohort consisting of 24 probands/families who tested negative for mutations in the CCM1, CCM2, and CCM3 genes by DNA sequence analysis. Surprisingly, the common CCM2 deletion spanning exons 2-10 is not present in this population. Further analysis of the Italian cohort by multiplex ligation-dependent probe analysis identified a total of ten deletions and one duplication. The overall spectrum of genomic rearrangements in the Italian cohort is thus quite different than that seen in a US cohort. These results suggest that there are elements within all three of the CCM genes that predispose them to large deletion/duplication events but that the common deletion spanning CCM2 exons 2-10 appears to be specific to the US population due to a founder effect.

Authors
Liquori, CL; Penco, S; Gault, J; Leedom, TP; Tassi, L; Esposito, T; Awad, IA; Frati, L; Johnson, EW; Squitieri, F; Marchuk, DA; Gianfrancesco, F
MLA Citation
Liquori, CL, Penco, S, Gault, J, Leedom, TP, Tassi, L, Esposito, T, Awad, IA, Frati, L, Johnson, EW, Squitieri, F, Marchuk, DA, and Gianfrancesco, F. "Different spectra of genomic deletions within the CCM genes between Italian and American CCM patient cohorts." Neurogenetics 9.1 (February 2008): 25-31.
PMID
18060436
Source
pubmed
Published In
Neurogenetics
Volume
9
Issue
1
Publish Date
2008
Start Page
25
End Page
31
DOI
10.1007/s10048-007-0109-x

Childhood socioeconomic status and serotonin transporter gene polymorphism enhance cardiovascular reactivity to mental stress.

OBJECTIVE: To test the hypothesis that low socioeconomic status (SES) and the 5HTTLPR L allele are associated with increased cardiovascular reactivity (CVR) to stress in a larger sample and that SES and 5HTTLPR genotypes interact to enhance CVR to stress. CVR to mental stress has been proposed as one mechanism linking stress to the pathogenesis of cardiovascular disease. The more transcriptionally efficient long (L) allele of a polymorphism of the serotonin transporter gene promoter (5HTTLPR) has been found associated with increased risk of myocardial infarction. We found the long allele associated with larger CVR to mental stress in a preliminary study of 54 normal volunteers. METHODS: Subjects included 165 normal community volunteers stratified for race, gender, and SES, who underwent mental stress testing. RESULTS: Childhood SES as indexed by Father's Education Level was associated with larger systolic blood pressure (SBP) (p < .05) and diastolic blood pressure (DBP) (p = .01) responses to mental stress. The L allele was associated with larger SBP (p = .04), DBP (p < .0001), and heart rate (p = .04) responses to mental stress compared with the short (S) allele. Subjects with the SS genotype and high Father's Education exhibited smaller SBP (5.2 mm Hg) and DBP (2.9 mm Hg) responses than subjects with LL genotype and low Father's Education (SBP = 13.3 mm Hg, p = .002; DBP = 9.7 mm Hg, p < .0001). CONCLUSIONS: Both the 5HTTLPR long allele and low SES, particularly during childhood, are associated with increased CVR to mental stress, which could account, at least in part, for the increased cardiovascular disease risk associated with these characteristics. If confirmed in further research, these characteristics could be used to identify persons who might benefit from preventive interventions.

Authors
Williams, RB; Marchuk, DA; Siegler, IC; Barefoot, JC; Helms, MJ; Brummett, BH; Surwit, RS; Lane, JD; Kuhn, CM; Gadde, KM; Ashley-Koch, A; Svenson, IK; Schanberg, SM
MLA Citation
Williams, RB, Marchuk, DA, Siegler, IC, Barefoot, JC, Helms, MJ, Brummett, BH, Surwit, RS, Lane, JD, Kuhn, CM, Gadde, KM, Ashley-Koch, A, Svenson, IK, and Schanberg, SM. "Childhood socioeconomic status and serotonin transporter gene polymorphism enhance cardiovascular reactivity to mental stress." Psychosom Med 70.1 (January 2008): 32-39.
PMID
18158371
Source
pubmed
Published In
Psychosomatic Medicine
Volume
70
Issue
1
Publish Date
2008
Start Page
32
End Page
39
DOI
10.1097/PSY.0b013e31815f66c3

Genetic considerations relevant to intracranial hemorrhage and brain arteriovenous malformations.

Brain arteriovenous malformations (AVMs) cause intracranial hemorrhage (ICH), especially in young adults. Molecular characterization of lesional tissue provides evidence for involvement of both angiogenic and inflammatory pathways, but the pathogenesis remains obscure and medical therapy is lacking. Abnormal expression patterns have been observed for proteins related to angiogenesis (e.g., vascular endothelial growth factor, angiopoietin-2, matrix metalloproteinase-9), and inflammation (e.g., interleukin-6 [IL-6] and myeloperoxidase). Macrophage and neutrophil invasion have also been observed in the absence of prior ICH. Candidate gene association studies have identified a number of germline variants associated with clinical ICH course and AVM susceptibility. A single nucleotide polymorphism (SNP) in activin receptor-like kinase-1 (ALK-1) is associated with AVM susceptibility, and SNPs in IL-6, tumor necrosis factor-alpha (TNF-alpha), and apolipoprotein-E (APOE) are associated with AVM rupture. These observations suggest that even without a complete understanding of the determinants of AVM development, the recent discoveries of downstream derangements in vascular function and integrity may offer potential targets for therapy development. Further, biomarkers can now be established for assessing ICH risk. These data will generate hypotheses that can be tested mechanistically in model systems, including surrogate phenotypes, such as vascular dysplasia and/or models recapitulating the clinical syndrome of recurrent spontaneous ICH.

Authors
Kim, H; Marchuk, DA; Pawlikowska, L; Chen, Y; Su, H; Yang, GY; Young, WL
MLA Citation
Kim, H, Marchuk, DA, Pawlikowska, L, Chen, Y, Su, H, Yang, GY, and Young, WL. "Genetic considerations relevant to intracranial hemorrhage and brain arteriovenous malformations." Acta Neurochir Suppl 105 (2008): 199-206. (Review)
PMID
19066109
Source
pubmed
Published In
Acta neurochirurgica. Supplementum
Volume
105
Publish Date
2008
Start Page
199
End Page
206

Neonatal co-infection with helicobacter species markedly accelerates the development of inflammation-associated colonic neoplasia in IL-10(-/-) mice.

BACKGROUND: Inflammatory bowel disease (IBD) is hypothesized to represent an aberrant immune response against enteric bacteria that occurs in a genetically susceptible host. Humans and mice with IBD are at markedly increased risk for colonic neoplasia. However, the long lead time required before development of inflammation-associated colon neoplasia in commonly used murine models of IBD slows the development of effective chemopreventative therapies. MATERIALS AND METHODS: Neonatal coinfection with Helicobacter typhlonius and Helicobacter rodentium was used to trigger the onset of IBD in mice deficient in the immunoregulatory cytokine interleukin (IL)-10. The severity of colon inflammation and incidence of neoplasia was determined histologically. RESULTS: IL-10(-/-) mice demonstrated early onset, severe colon inflammation following neonatal infection with H. typhlonius and H. rodentium. The incidence of inflammation-associated colon neoplasia was approximately 95% at a mean age of 21 +/- 2 weeks. Mutation of endoglin, an accessory receptor for TGF-beta, did not affect the severity of IBD or the incidence of neoplasia in this model. CONCLUSIONS: The rapid onset of severe colon inflammation and multiple neoplastic lesions in the colons of IL-10(-/-) mice neonatally coinfected with H. typhlonius and H. rodentium makes this model well-suited for investigating the mechanisms involved in inflammation-associated colon cancer as well as its chemoprevention.

Authors
Hale, LP; Perera, D; Gottfried, MR; Maggio-Price, L; Srinivasan, S; Marchuk, D
MLA Citation
Hale, LP, Perera, D, Gottfried, MR, Maggio-Price, L, Srinivasan, S, and Marchuk, D. "Neonatal co-infection with helicobacter species markedly accelerates the development of inflammation-associated colonic neoplasia in IL-10(-/-) mice." Helicobacter 12.6 (December 2007): 598-604.
PMID
18001399
Source
pubmed
Published In
Helicobacter
Volume
12
Issue
6
Publish Date
2007
Start Page
598
End Page
604
DOI
10.1111/j.1523-5378.2007.00552.x

Hypertension and albuminuria in chronic kidney disease mapped to a mouse chromosome 11 locus.

Chronic kidney disease (CKD) is a key cause of hypertension and a potent independent risk for cardiovascular disease. Epidemiological studies suggest a strong genetic component determining susceptibility for renal disease and, by inference, the associated cardiovascular risk. With a subtotal nephrectomy model of kidney disease, we found the 129S6 mouse strain to be very susceptible to the development of hypertension, albuminuria, and kidney injury, whereas the C57BL/6 strain is relatively resistant. Accordingly, we set out to map quantitative trait loci conferring susceptibility to hypertension and albuminuria using this model with F2 mice. We found significant linkage of the blood pressure trait to two loci. At D11Mit143, mice homozygous for the 129S6 allele had significantly higher systolic blood pressure than mice heterozygous or homozygous for the C57BL/6 allele. Similarly, at D1Mit308, there was an excellent correlation between genotype and the blood pressure phenotype. The effect of the chromosome 11 locus was verified with a separate cohort of F2 mice. For the albuminuria trait, a significant locus was found at D11Mit143, which overlaps the blood pressure trait locus. Our studies have identified a region spanning approximately 8 cM on mouse chromosome 11 that is associated with susceptibility to hypertension and albuminuria in CKD.

Authors
Salzler, HR; Griffiths, R; Ruiz, P; Chi, L; Frey, C; Marchuk, DA; Rockman, HA; Le, TH
MLA Citation
Salzler, HR, Griffiths, R, Ruiz, P, Chi, L, Frey, C, Marchuk, DA, Rockman, HA, and Le, TH. "Hypertension and albuminuria in chronic kidney disease mapped to a mouse chromosome 11 locus." Kidney Int 72.10 (November 2007): 1226-1232.
PMID
17851470
Source
pubmed
Published In
Kidney international
Volume
72
Issue
10
Publish Date
2007
Start Page
1226
End Page
1232
DOI
10.1038/sj.ki.5002519

Highly variable penetrance in subjects affected with cavernous cerebral angiomas (CCM) carrying novel CCM1 and CCM2 mutations.

Cavernous vascular malformations may affect brain and out-of-brain tissues. In most cases, cerebral cavernous malformations (CCMs) involve the brain alone, and are rarely associated with skin hemangiomas, spinal cord, retinal, hepatic or vertebral lesions. CCMs can cause seizures, intracranial and spinal haemorrhages, focal neurological deficits, and migraine-like headaches. After collecting CCM families of Italian origin and investigating the genetic basis of the disorder we disclosed two novel molecular variations in the KRIT1 and MGC4607 genes. We found a novel CCM1 gene mutation (Q66X) in a family with apparently asymptomatic old-aged mutation carriers and patients who either had skin angiomas alone or the full association of cerebral, spinal, and skin lesions. In this family we report the highest variability in mutation penetrance so far described, including the presence of CCM in one subject since birth (surgery at 19 months of age), a condition to our knowledge so far unreported. In a CCM2 affected family, we also report a novel causative mutation, (54_55delAC) in exon 2 of the MGC4607 gene, that produces a truncated protein containing only 22 amino acids. These data describe novel CCM mutations associated with a particularly high variability of the penetrance causing, in some cases, reduced expression of clinical symptoms and sporadic cases with apparent negative family history. Hence they emphasize the importance of DNA-based diagnostics and genetic counseling to identify unaffected mutation carriers subjects, even at advanced age.

Authors
Gianfrancesco, F; Cannella, M; Martino, T; Maglione, V; Esposito, T; Innocenzi, G; Vitale, E; Liquori, CL; Marchuk, DA; Squitieri, F
MLA Citation
Gianfrancesco, F, Cannella, M, Martino, T, Maglione, V, Esposito, T, Innocenzi, G, Vitale, E, Liquori, CL, Marchuk, DA, and Squitieri, F. "Highly variable penetrance in subjects affected with cavernous cerebral angiomas (CCM) carrying novel CCM1 and CCM2 mutations." Am J Med Genet B Neuropsychiatr Genet 144B.5 (July 5, 2007): 691-695.
PMID
17440989
Source
pubmed
Published In
American Journal of Medical Genetics Part B: Neuropsychiatric Genetics
Volume
144B
Issue
5
Publish Date
2007
Start Page
691
End Page
695
DOI
10.1002/ajmg.b.30381

Arteriovenous malformation.

Authors
Young, WL; Kwok, P-Y; Pawlikowska, L; Lawton, MT; Kim, H; Hysi, PG; Marchuk, DA
MLA Citation
Young, WL, Kwok, P-Y, Pawlikowska, L, Lawton, MT, Kim, H, Hysi, PG, and Marchuk, DA. "Arteriovenous malformation." J Neurosurg 106.4 (April 2007): 731-732. (Letter)
PMID
17432733
Source
pubmed
Published In
Journal of neurosurgery
Volume
106
Issue
4
Publish Date
2007
Start Page
731
End Page
732
DOI
10.3171/jns.2007.106.4.731

Deletions in CCM2 are a common cause of cerebral cavernous malformations.

Cerebral cavernous malformations (CCMs) are vascular abnormalities of the brain that can result in a variety of neurological disabilities, including hemorrhagic stroke and seizures. Mutations in the gene KRIT1 are responsible for CCM1, mutations in the gene MGC4607 are responsible for CCM2, and mutations in the gene PDCD10 are responsible for CCM3. DNA sequence analysis of the known CCM genes in a cohort of 63 CCM-affected families showed that a high proportion (40%) of these lacked any identifiable mutation. We used multiplex ligation-dependent probe analysis to screen 25 CCM1, -2, and -3 mutation-negative probands for potential deletions or duplications within all three CCM genes. We identified a total of 15 deletions: 1 in the CCM1 gene, 0 in the CCM3 gene, and 14 in the CCM2 gene. In our cohort, mutation screening that included sequence and deletion analyses gave disease-gene frequencies of 40% for CCM1, 38% for CCM2, 6% for CCM3, and 16% with no mutation detected. These data indicate that the prevalence of CCM2 is much higher than previously predicted, nearly equal to CCM1, and that large genomic deletions in the CCM2 gene represent a major component of this disease. A common 77.6-kb deletion spanning CCM2 exons 2-10 was identified, which is present in 13% of our entire CCM cohort. Eight probands exhibit an apparently identical recombination event in the CCM2 gene, involving an AluSx in intron 1 and an AluSg distal to exon 10. Haplotype analysis revealed that this CCM2 deletion occurred independently at least twice in our families. We hypothesize that these deletions occur in a hypermutable region because of surrounding repetitive sequence elements that may catalyze the formation of intragenic deletions.

Authors
Liquori, CL; Berg, MJ; Squitieri, F; Leedom, TP; Ptacek, L; Johnson, EW; Marchuk, DA
MLA Citation
Liquori, CL, Berg, MJ, Squitieri, F, Leedom, TP, Ptacek, L, Johnson, EW, and Marchuk, DA. "Deletions in CCM2 are a common cause of cerebral cavernous malformations." Am J Hum Genet 80.1 (January 2007): 69-75.
PMID
17160895
Source
pubmed
Published In
The American Journal of Human Genetics
Volume
80
Issue
1
Publish Date
2007
Start Page
69
End Page
75
DOI
10.1086/510439

Reply

Authors
Donahue, MP; Marchuk, DA; Rockman, HA
MLA Citation
Donahue, MP, Marchuk, DA, and Rockman, HA. "Reply." Journal of the American College of Cardiology 49.10 (2007): 1106-1107.
Source
scival
Published In
JACC - Journal of the American College of Cardiology
Volume
49
Issue
10
Publish Date
2007
Start Page
1106
End Page
1107
DOI
10.1016/j.jacc.2006.12.019

Genetic analysis of a family with hereditary glomuvenous malformations

Glomuvenous malformations (MIM 138000) are rare vascular malformations consisting of glomus cells, and in affected individuals, lesions may appear in any number anywhere on the body. We analysed the DNA of one family with hereditary glomuvenous malformations and identified the mutation causing the disease in the glomulin gene on chromosome 1 p22. The deletion started at base pair 157: 157delAAGAA, which is a deletion of five base pairs. This mutation has been found in Europe, the USA and Australia, suggesting a founder effect with common ancestry. Thus far, no second-hit mutation for the 157delAAGAA mutation has been identified. © 2007 The Authors.

Authors
Ostberg, A; Moreno, G; Su, T; Trisnowati, N; Marchuk, D; Murrell, D
MLA Citation
Ostberg, A, Moreno, G, Su, T, Trisnowati, N, Marchuk, D, and Murrell, D. "Genetic analysis of a family with hereditary glomuvenous malformations." Australasian Journal of Dermatology 48.3 (2007): 170-173.
Source
scival
Published In
Australasian Journal of Dermatology
Volume
48
Issue
3
Publish Date
2007
Start Page
170
End Page
173
DOI
10.1111/j.1440-0960.2007.00373.x

Erratum: Genetic analysis of a family with hereditary glomuvenous malformations (Australasian Journal of Dermatology (2007) 48, (170-173))

Authors
Ostberg, A; Moreno, G; Su, T; Trisnowati, N; Marchuk, D; Murrell, D
MLA Citation
Ostberg, A, Moreno, G, Su, T, Trisnowati, N, Marchuk, D, and Murrell, D. "Erratum: Genetic analysis of a family with hereditary glomuvenous malformations (Australasian Journal of Dermatology (2007) 48, (170-173))." Australasian Journal of Dermatology 48.4 (2007): 261--.
Source
scival
Published In
Australasian Journal of Dermatology
Volume
48
Issue
4
Publish Date
2007
Start Page
261-
DOI
10.1111/j.1440-0960.2007.00407.x

No evidence for maternal-fetal microchimerism in infantile hemangioma: a molecular genetic investigation.

In this study, using the placental origin theory as a basis, we set out to explore whether hemangioma endothelial cells (HEC) were maternal in origin. We rigorously addressed this hypothesis using several molecular genetic techniques. Fluorescent in situ hybridization on surgical specimens of proliferating hemangiomas (n=8) demonstrated no XX-labeled HEC from resected tumors of male infants. This analysis was followed by PCR genotyping of HEC (n=11) using microsatellite markers where cellular components were genotyped and compared to genomic DNA of corresponding mother-child pairs. In the seven informative mother-child pairs, HEC matched the genotype of the child and not the maternal genotype. Concerned that HEC represented a mixed population of cells, we subsequently enriched for cells using the placental-specific endothelial cell (EC) marker, Fc gammaRII. Three informative mother-child pairs exhibited only the genotype of the child in our enriched cell population. Using sequence analysis, we identified an informative single nucleotide polymorphism in an exon of the placental-EC-specific protein, GLUT1. When comparing GLUT1 complementary DNA (cDNA) with mother-child DNA, the genotype of the cDNA matched the constitutional DNA of the child. Our results indicate that hemangiomas are not microchimeric in origin. This study provides further insight into the origin of a tumor whose pathogenesis remains elusive.

Authors
Pittman, KM; Losken, HW; Kleinman, ME; Marcus, JR; Blei, F; Gurtner, GC; Marchuk, DA
MLA Citation
Pittman, KM, Losken, HW, Kleinman, ME, Marcus, JR, Blei, F, Gurtner, GC, and Marchuk, DA. "No evidence for maternal-fetal microchimerism in infantile hemangioma: a molecular genetic investigation." J Invest Dermatol 126.11 (November 2006): 2533-2538.
PMID
16902414
Source
pubmed
Published In
Journal of Investigative Dermatology
Volume
126
Issue
11
Publish Date
2006
Start Page
2533
End Page
2538
DOI
10.1038/sj.jid.5700516

Redefining heart failure: the utility of genomics.

In this era of genomics, new technologies and the information that they generate have a wide range of potential applications to heart failure. Though there has not been widespread practical use of genomic information in everyday practice, there are many examples of how this information is beginning to transform the way we look at disease states in terms of diagnosis, prognosis, and treatment. The experience of oncology and other fields helps inform the heart failure field of not only the use of this information in investigating diagnosis, prognosis, and treatment response, but the reciprocal nature of this information. This information can be clinically useful (for instance, predicting treatment response) as well as further drive laboratory investigation (teasing out the biological pathways in non-responders to treatment can be a focus of new drug discovery); this is the essence of translational medicine. We believe that this is a good time to review where new technologies and information they generate can be placed into our classic understanding of heart failure: that is how we might redefine cardiomyopathy given our new information. Here we will review genomic evidence to date and how it can and may be considered in the evaluation and management of cardiomyopathies.

Authors
Donahue, MP; Marchuk, DA; Rockman, HA
MLA Citation
Donahue, MP, Marchuk, DA, and Rockman, HA. "Redefining heart failure: the utility of genomics." J Am Coll Cardiol 48.7 (October 3, 2006): 1289-1298. (Review)
PMID
17010784
Source
pubmed
Published In
Journal of the American College of Cardiology
Volume
48
Issue
7
Publish Date
2006
Start Page
1289
End Page
1298
DOI
10.1016/j.jacc.2006.05.062

SMAD4 mutations found in unselected HHT patients.

BACKGROUND: Hereditary haemorrhagic telangiectasia (HHT) is an autosomal dominant disease exhibiting multifocal vascular telangiectases and arteriovenous malformations. The majority of cases are caused by mutations in either the endoglin (ENG) or activin receptor-like kinase 1 (ALK1, ACVRL1) genes; both members of the transforming growth factor (TGF)-beta pathway. Mutations in SMAD4, another TGF-beta pathway member, are seen in patients with the combined syndrome of juvenile polyposis (JP) and HHT (JP-HHT). METHODS: We sought to determine if HHT patients without any apparent history of JP, who were undergoing routine diagnostic testing, would have mutations in SMAD4. We tested 30 unrelated HHT patients, all of whom had been referred for DNA based testing for HHT and were found to be negative for mutations in ENG and ALK1. RESULTS: Three of these people harboured mutations in SMAD4, a rate of 10% (3/30). The SMAD4 mutations were similar to those found in other patients with the JP-HHT syndrome. CONCLUSIONS: The identification of SMAD4 mutations in HHT patients without prior diagnosis of JP has significant and immediate clinical implications, as these people are likely to be at risk of having JP-HHT with the associated increased risk of gastrointestinal cancer. We propose that routine DNA based testing for HHT should include SMAD4 for samples in which mutations in neither ENG nor ALK1 are identified. HHT patients with SMAD4 mutations should be screened for colonic and gastric polyps associated with JP.

Authors
Gallione, CJ; Richards, JA; Letteboer, TGW; Rushlow, D; Prigoda, NL; Leedom, TP; Ganguly, A; Castells, A; Ploos van Amstel, JK; Westermann, CJJ; Pyeritz, RE; Marchuk, DA
MLA Citation
Gallione, CJ, Richards, JA, Letteboer, TGW, Rushlow, D, Prigoda, NL, Leedom, TP, Ganguly, A, Castells, A, Ploos van Amstel, JK, Westermann, CJJ, Pyeritz, RE, and Marchuk, DA. "SMAD4 mutations found in unselected HHT patients." J Med Genet 43.10 (October 2006): 793-797.
PMID
16613914
Source
pubmed
Published In
Journal of medical genetics
Volume
43
Issue
10
Publish Date
2006
Start Page
793
End Page
797
DOI
10.1136/jmg.2006.041517

Precocious osteoarthritis in a family with recurrent COL2A1 mutation.

OBJECTIVE: To examine the genotypic and phenotypic characteristics of a Micronesian kindred with autosomal dominant precocious osteoarthritis (OA). METHODS: We reviewed records and radiographs of 3 index patients and their parents, administered questionnaires to 16 additional kindred members, performed whole-genome scans of 24 family members, and sequenced relevant genes from 16 family members. RESULTS: The kindred displayed early onset OA, enlarged epiphyses, platyspondyly, and brachydactyly with dysplastic findings consistent with mild spondyloepiphyseal dysplasia. Genetic analysis revealed an arginine to cysteine substitution at position 75 of the collagen 2A1 gene, a mutation that has been described in 4 other geographically distinct families. The major phenotypic differences among the families were in height (ranging from short to tall) and hearing loss noted in 3 of the 5 families. CONCLUSION: The presence of the COL2A1 Arg75Cys mutation in 5 geographically distinct areas helps to confirm a potential mutational hotspot. The diverse phenotypic spectrum suggests that modifier genes and environmental factors play a role in the expression of this mutation.

Authors
Carlson, KM; Yamaga, KM; Reinker, KA; Hsia, YE; Carpenter, C; Abe, LM; Perry, AK; Person, DA; Marchuk, DA; Raney, EM
MLA Citation
Carlson, KM, Yamaga, KM, Reinker, KA, Hsia, YE, Carpenter, C, Abe, LM, Perry, AK, Person, DA, Marchuk, DA, and Raney, EM. "Precocious osteoarthritis in a family with recurrent COL2A1 mutation." J Rheumatol 33.6 (June 2006): 1133-1136.
PMID
16755660
Source
pubmed
Published In
The Journal of rheumatology
Volume
33
Issue
6
Publish Date
2006
Start Page
1133
End Page
1136

A new locus for dominant hereditary spastic paraplegia maps to chromosome 2p12.

Authors
Züchner, S; Kail, ME; Nance, MA; Gaskell, PC; Svenson, IK; Marchuk, DA; Pericak-Vance, MA; Ashley-Koch, AE
MLA Citation
Züchner, S, Kail, ME, Nance, MA, Gaskell, PC, Svenson, IK, Marchuk, DA, Pericak-Vance, MA, and Ashley-Koch, AE. "A new locus for dominant hereditary spastic paraplegia maps to chromosome 2p12." Neurogenetics 7.2 (May 2006): 127-129. (Letter)
PMID
16565863
Source
pubmed
Published In
Neurogenetics
Volume
7
Issue
2
Publish Date
2006
Start Page
127
End Page
129
DOI
10.1007/s10048-006-0029-1

Apolipoprotein E epsilon 2 is associated with new hemorrhage risk in brain arteriovenous malformations.

OBJECTIVE: Patients with brain arteriovenous malformation (AVM) are at life-threatening risk of intracranial hemorrhage (ICH). Identification of genetic variants associated with increased new ICH risk would facilitate risk stratification and guide therapeutic intervention. METHODS: Brain AVM patients evaluated at University of California, San Francisco or Kaiser Permanente Northern California were followed longitudinally. Primary outcome was new ICH after diagnosis; censoring events were any AVM treatment or last follow-up examination. The association of ApoE epsilon2 and epsilon4 genotype with new ICH was evaluated by Kaplan-Meier survival analysis and further characterized via a Cox proportional hazards model. RESULTS: We genotyped 284 brain AVM patients (50% women; 57% Caucasian; median follow-up time, 0.3 yr) including 18 patients with a history of new ICH). ApoE epsilon2, but not ApoE epsilon4 genotype, was associated with new ICH (P = 0.0052). ApoE epsilon2 carriers had fivefold increased risk of new ICH (hazard ratio, 5.09; 95% confidence interval, 1.46-17.7; P = 0.010; Cox proportional hazards model adjusting for race/ethnicity and clinical presentation). Subset analysis in the largest homogenous ethnic subcohort (Caucasians) confirmed the increased risk of new ICH in ApoE epsilon2 carriers (hazard ratio, 8.71; 95% confidence interval, 1.4-53.9; P = 0.020; multivariate model adjusting for clinical presentation). CONCLUSION: ApoE genotype may influence the risk of ICH in the natural course of brain AVM. The identification of genetic predictors of ICH risk may facilitate estimation of AVM natural history risk and individualize clinical decision-making and therapeutic recommendations.

Authors
Pawlikowska, L; Poon, KYT; Achrol, AS; McCulloch, CE; Ha, C; Lum, K; Zaroff, JG; Ko, NU; Johnston, SC; Sidney, S; Marchuk, DA; Lawton, MT; Kwok, P-Y; Young, WL
MLA Citation
Pawlikowska, L, Poon, KYT, Achrol, AS, McCulloch, CE, Ha, C, Lum, K, Zaroff, JG, Ko, NU, Johnston, SC, Sidney, S, Marchuk, DA, Lawton, MT, Kwok, P-Y, and Young, WL. "Apolipoprotein E epsilon 2 is associated with new hemorrhage risk in brain arteriovenous malformations." Neurosurgery 58.5 (May 2006): 838-843.
PMID
16639317
Source
pubmed
Published In
Neurosurgery
Volume
58
Issue
5
Publish Date
2006
Start Page
838
End Page
843
DOI
10.1227/01.NEU.0000209605.18358.E5

ALK1 signalling analysis identifies angiogenesis related genes and reveals disparity between TGF-beta and constitutively active receptor induced gene expression.

BACKGROUND: TGF-beta1 is an important angiogenic factor involved in the different aspects of angiogenesis and vessel maintenance. TGF-beta signalling is mediated by the TbetaRII/ALK5 receptor complex activating the Smad2/Smad3 pathway. In endothelial cells TGF-beta utilizes a second type I receptor, ALK1, activating the Smad1/Smad5 pathway. Consequently, a perturbance of ALK1, ALK5 or TbetaRII activity leads to vascular defects. Mutations in ALK1 cause the vascular disorder hereditary hemorrhagic telangiectasia (HHT). METHODS: The identification of ALK1 and not ALK5 regulated genes in endothelial cells, might help to better understand the development of HHT. Therefore, the human microvascular endothelial cell line HMEC-1 was infected with a recombinant constitutively active ALK1 adenovirus, and gene expression was studied by using gene arrays and quantitative real-time PCR analysis. RESULTS: After 24 hours, 34 genes were identified to be up-regulated by ALK1 signalling. Analysing ALK1 regulated gene expression after 4 hours revealed 13 genes to be up- and 2 to be down-regulated. Several of these genes, including IL-8, ET-1, ID1, HPTPeta and TEAD4 are reported to be involved in angiogenesis. Evaluation of ALK1 regulated gene expression in different human endothelial cell types was not in complete agreement. Further on, disparity between constitutively active ALK1 and TGF-beta1 induced gene expression in HMEC-1 cells and primary HUVECs was observed. CONCLUSION: Gene array analysis identified 49 genes to be regulated by ALK1 signalling and at least 14 genes are reported to be involved in angiogenesis. There was substantial agreement between the gene array and quantitative real-time PCR data. The angiogenesis related genes might be potential HHT modifier genes. In addition, the results suggest endothelial cell type specific ALK1 and TGF-beta signalling.

Authors
Lux, A; Salway, F; Dressman, HK; Kröner-Lux, G; Hafner, M; Day, PJR; Marchuk, DA; Garland, J
MLA Citation
Lux, A, Salway, F, Dressman, HK, Kröner-Lux, G, Hafner, M, Day, PJR, Marchuk, DA, and Garland, J. "ALK1 signalling analysis identifies angiogenesis related genes and reveals disparity between TGF-beta and constitutively active receptor induced gene expression. (Published online)" BMC Cardiovasc Disord 6 (April 4, 2006): 13-.
PMID
16594992
Source
pubmed
Published In
BMC Cardiovascular Disorders
Volume
6
Publish Date
2006
Start Page
13
DOI
10.1186/1471-2261-6-13

Genomic characterization of POS5, the Saccharomyces cerevisiae mitochondrial NADH kinase.

Disruption of the Saccharomyces cerevisiae mitochondrial NADH kinase POS5 increases the mitochondrial mutation rate 50-fold. Whereas most multicellular eukaryotic genomes have one NADH kinase gene, the yeast genome contains three distinct genes encoding NAD/H kinase activity. To determine if all three genes are essential for viability we constructed combinations of gene knockouts. We show that only the pos5Deltautr1Delta combination is synthetically lethal, demonstrating an essential overlapping function, and showing that NAD/H kinase activity is essential for eukaryotic viability. The single human NAD/H kinase gene can rescue the lethality of the double knockout in yeast, demonstrating that the single human gene can fill the various functions provided by the three yeast genes. The human NAD/H kinase gene harbors very common sequence variants, but all of these equally complement the synthetic lethality in yeast, illustrating that each of these are functionally wild-type. To understand the molecular mechanism of the mitochondrial genome instability of pos5 mutation we performed gene expression analysis on the pos5Delta. The pos5Delta resulted in an increase in expression of most of the iron transport genes including key genes involved in iron-sulfur cluster assembly. Decreased expression occurred in many genes involved in the electron transport chain. We show that the pos5Delta expression pattern is similar to the frataxin homolog knockout (yfh1Delta), the yeast model for Friedreich's ataxia. These combined data show that the POS5 NAD/H kinase is an important protein required for a variety of essential cellular pathways and that deficient iron-sulfur cluster assembly may play a critical role in the mitochondrial mutator phenotype observed in the pos5Delta.

Authors
Shianna, KV; Marchuk, DA; Strand, MK
MLA Citation
Shianna, KV, Marchuk, DA, and Strand, MK. "Genomic characterization of POS5, the Saccharomyces cerevisiae mitochondrial NADH kinase." Mitochondrion 6.2 (April 2006): 94-101.
PMID
16621727
Source
pubmed
Published In
Mitochondrion
Volume
6
Issue
2
Publish Date
2006
Start Page
94
End Page
101
DOI
10.1016/j.mito.2006.02.003

Neuronal expression of the Ccm2 gene in a new mouse model of cerebral cavernous malformations.

Cerebral cavernous malformations are vascular defects of the central nervous system consisting of clusters of dilated vessels that are subject to frequent hemorrhaging. The genes mutated in three forms of autosomal dominant cerebral cavernous malformations have been cloned, but it remains unclear which cell type is ultimately responsible for the lesion. In this article we describe mice with a gene trap insertion in the Ccm2 gene. Consistent with the human phenotype, heterozygous animals develop cerebral vascular malformations, although penetrance is low. Beta-galactosidase activity in heterozygous brain and in situ hybridization in wild-type brain revealed Ccm2 expression in neurons and choroid plexus but not in vascular endothelium of small vessels in the brain. The expression pattern of Ccm2 is similar to that of the Ccm1 gene and its interacting protein ICAP1 (Itgb1bp1). These data suggest that cerebral cavernous malformations arise as a result of defects in the neural parenchyma surrounding the vascular endothelial cells in the brain.

Authors
Plummer, NW; Squire, TL; Srinivasan, S; Huang, E; Zawistowski, JS; Matsunami, H; Hale, LP; Marchuk, DA
MLA Citation
Plummer, NW, Squire, TL, Srinivasan, S, Huang, E, Zawistowski, JS, Matsunami, H, Hale, LP, and Marchuk, DA. "Neuronal expression of the Ccm2 gene in a new mouse model of cerebral cavernous malformations." Mamm Genome 17.2 (February 2006): 119-128.
PMID
16465592
Source
pubmed
Published In
Mammalian Genome
Volume
17
Issue
2
Publish Date
2006
Start Page
119
End Page
128
DOI
10.1007/s00335-005-0098-8

Low frequency of PDCD10 mutations in a panel of CCM3 probands: potential for a fourth CCM locus.

Cerebral cavernous malformations (CCMs) are vascular abnormalities of the brain that can result in a variety of neurological disabilities, including stroke and seizures. Linkage analyses using autosomal dominant families manifesting CCMs have identified three different causative loci on chromosomes 7q21.2 (CCM1), 7p13 (CCM2), and 3q25.2-q27 (CCM3). Mutations in the gene Krit1 are responsible for CCM1, mutations in the gene MGC4607 are responsible for CCM2, and mutations in the gene PDCD10 were recently reported to be responsible for CCM3. We report here that sequence analysis of PDCD10 in a panel of 29 probands lacking Krit1 and MGC4607 mutations revealed only three mutations. The frequency of identified mutations in the PDCD10 gene was surprisingly low, especially given that this panel was heavily biased towards non-CCM1, non-CCM2 probands. These data are in stark contrast with the linkage data, which suggests that 40% of inherited cases would be due to mutations in this gene. Interestingly, when examining the haplotypes of previously published CCM3 families, we found a distinct recombination event in one of the largest CCM3 families that excludes the PDCD10 gene. Although there are many potential explanations for this observation, when combined with the apparent under-representation of causative CCM mutations in PDCD10, this recombination event in a CCM3-linked family suggests that there may be an additional CCM gene in the same chromosomal region.

Authors
Liquori, CL; Berg, MJ; Squitieri, F; Ottenbacher, M; Sorlie, M; Leedom, TP; Cannella, M; Maglione, V; Ptacek, L; Johnson, EW; Marchuk, DA
MLA Citation
Liquori, CL, Berg, MJ, Squitieri, F, Ottenbacher, M, Sorlie, M, Leedom, TP, Cannella, M, Maglione, V, Ptacek, L, Johnson, EW, and Marchuk, DA. "Low frequency of PDCD10 mutations in a panel of CCM3 probands: potential for a fourth CCM locus." Hum Mutat 27.1 (January 2006): 118-.
PMID
16329096
Source
pubmed
Published In
Human Mutation
Volume
27
Issue
1
Publish Date
2006
Start Page
118
DOI
10.1002/humu.9389

Tumor necrosis factor-alpha-238G>A promoter polymorphism is associated with increased risk of new hemorrhage in the natural course of patients with brain arteriovenous malformations.

BACKGROUND AND PURPOSE: Identification of single-nucleotide polymorphisms (SNPs) associated with increased risk of new intracranial hemorrhage (ICH) after brain arteriovenous malformation (BAVM) diagnosis would facilitate risk stratification and identify potential targets for therapeutic intervention. METHODS: Patients with BAVM were longitudinally followed. Primary outcome was new ICH after diagnosis; censoring events were last follow-up or any BAVM treatment. We genotyped 4 promoter SNPs in 2 inflammatory cytokine genes: interleukin-6 (IL-6-174G>C; IL-6-572G>C) and tumor necrosis factor-alpha (TNF-alpha-238G>A; TNF-alpha-308G>A). Association of genotype with risk of new ICH was screened using chi2; SNPs associated with new ICH were further characterized using Cox proportional hazards. RESULTS: We genotyped 280 patients (50% female; 59% white, mean+/-SD age at diagnosis 37+/-17 years; 40% presenting with ICH). TNF-alpha-238G>A was associated with increased risk of new ICH after diagnosis (chi2; P=0.003). After adjusting for age, race/ethnicity, and clinical presentation, the risk of new ICH was increased for patients with TNF-alpha-238 AG genotype (hazard ratio, 4.01; P=0.015). No other SNP was found to be associated with new ICH. CONCLUSIONS: A TNF-alpha SNP was associated with increased risk of new ICH in the natural course of BAVMs. The role of inflammatory cytokines in the pathogenesis of BAVM hemorrhage merits further study.

Authors
Achrol, AS; Pawlikowska, L; McCulloch, CE; Poon, KYT; Ha, C; Zaroff, JG; Johnston, SC; Lee, C; Lawton, MT; Sidney, S; Marchuk, DA; Kwok, P-Y; Young, WL; UCSF BAVM Study Project,
MLA Citation
Achrol, AS, Pawlikowska, L, McCulloch, CE, Poon, KYT, Ha, C, Zaroff, JG, Johnston, SC, Lee, C, Lawton, MT, Sidney, S, Marchuk, DA, Kwok, P-Y, Young, WL, and UCSF BAVM Study Project, . "Tumor necrosis factor-alpha-238G>A promoter polymorphism is associated with increased risk of new hemorrhage in the natural course of patients with brain arteriovenous malformations." Stroke 37.1 (January 2006): 231-234.
PMID
16322490
Source
pubmed
Published In
Stroke
Volume
37
Issue
1
Publish Date
2006
Start Page
231
End Page
234
DOI
10.1161/01.STR.0000195133.98378.4b

Genomic characterization of POS5, the Saccharomyces cerevisiae mitochondrial NADH kinase.

Disruption of the Saccharomyces cerevisiae mitochondrial NADH kinase POS5 increases the mitochondrial mutation rate 50-fold. Whereas most multicellular eukaryotic genomes have one NADH kinase gene, the yeast genome contains three distinct genes encoding NAD/H kinase activity. To determine if all three genes are essential for viability we constructed combinations of gene knockouts. We show that only the pos5Deltautr1Delta combination is synthetically lethal, demonstrating an essential overlapping function, and showing that NAD/H kinase activity is essential for eukaryotic viability. The single human NAD/H kinase gene can rescue the lethality of the double knockout in yeast, demonstrating that the single human gene can fill the various functions provided by the three yeast genes. The human NAD/H kinase gene harbors very common sequence variants, but all of these equally complement the synthetic lethality in yeast, illustrating that each of these are functionally wild-type. To understand the molecular mechanism of the mitochondrial genome instability of pos5 mutation we performed gene expression analysis on the pos5Delta. The pos5Delta resulted in an increase in expression of most of the iron transport genes including key genes involved in iron-sulfur cluster assembly. Decreased expression occurred in many genes involved in the electron transport chain. We show that the pos5Delta expression pattern is similar to the frataxin homolog knockout (yfh1Delta), the yeast model for Friedreich's ataxia. These combined data show that the POS5 NAD/H kinase is an important protein required for a variety of essential cellular pathways and that deficient iron-sulfur cluster assembly may play a critical role in the mitochondrial mutator phenotype observed in the pos5Delta.

Authors
Shianna, KV; Marchuk, DA; Strand, MK
MLA Citation
Shianna, KV, Marchuk, DA, and Strand, MK. "Genomic characterization of POS5, the Saccharomyces cerevisiae mitochondrial NADH kinase." Mitochondrion. 6.2 (2006): 94-101.
Source
scival
Published In
Mitochondrion
Volume
6
Issue
2
Publish Date
2006
Start Page
94
End Page
101

Molecular classification of patients with unexplained hamartomatous and hyperplastic polyposis.

CONTEXT: Significant proportions of patients with hamartomatous polyposis or with hyperplastic/mixed polyposis remain without specific clinical and molecular diagnosis or present atypically. Assigning a syndromic diagnosis is important because it guides management, especially surveillance and prophylactic surgery. OBJECTIVE: To systematically classify patients with unexplained hamartomatous or hyperplastic/mixed polyposis by extensive molecular analysis in the context of central rereview of histopathology results. DESIGN, SETTING, AND PATIENTS: Prospective, referral-based study of 49 unrelated patients from outside institutions (n = 28) and at a comprehensive cancer center (n = 21), conducted from May 2, 2002, until December 15, 2004. Germline analysis of PTEN, BMPR1A, STK11 (sequence, deletion), SMAD4, and ENG (sequence), specific exon screening of BRAF, MYH, and BHD, and rereview of polyp histology results were performed. MAIN OUTCOME MEASURES: Molecular, clinical, and histopathological findings in patients with unexplained polyposis. RESULTS: Of the 49 patients, 11 (22%) had germline mutations. Of 14 patients with juvenile polyposis, 2 with early-onset disease had mutations in ENG, encoding endoglin, previously only associated with hereditary hemorrhagic telangiectasia; 1 had hemizygous deletion encompassing PTEN and BMPR1A; and 1 had an SMAD4 mutation. One individual previously classified with Peutz-Jeghers syndrome had a PTEN deletion. Among 9 individuals with an unknown hamartomatous polyposis, 4 had mutations in STK11 (1), BMPR1A (2), and SMAD4 (1). Of the 23 patients with hyperplastic/mixed polyposis, 2 had PTEN mutations. Substantial discrepancies in histopathology results were seen. CONCLUSIONS: Systematic molecular classification of 49 patients with unexplained hamartomatous or hyperplastic polyposis uncovered a potential novel susceptibility gene, ENG, for juvenile polyposis. Importantly, given the substantial proportion of patients found to have germline mutations, more extensive analysis of the known susceptibility genes is indicated. Rereview of histology results by a dedicated gastrointestinal pathologist should be considered routinely, as organ-specific surveillance rests on defining syndromic diagnosis.

Authors
Sweet, K; Willis, J; Zhou, X-P; Gallione, C; Sawada, T; Alhopuro, P; Khoo, SK; Patocs, A; Martin, C; Bridgeman, S; Heinz, J; Pilarski, R; Lehtonen, R; Prior, TW; Frebourg, T; Teh, BT; Marchuk, DA; Aaltonen, LA; Eng, C
MLA Citation
Sweet, K, Willis, J, Zhou, X-P, Gallione, C, Sawada, T, Alhopuro, P, Khoo, SK, Patocs, A, Martin, C, Bridgeman, S, Heinz, J, Pilarski, R, Lehtonen, R, Prior, TW, Frebourg, T, Teh, BT, Marchuk, DA, Aaltonen, LA, and Eng, C. "Molecular classification of patients with unexplained hamartomatous and hyperplastic polyposis." JAMA 294.19 (November 16, 2005): 2465-2473.
PMID
16287957
Source
pubmed
Published In
JAMA : the journal of the American Medical Association
Volume
294
Issue
19
Publish Date
2005
Start Page
2465
End Page
2473
DOI
10.1001/jama.294.19.2465

CCM1 and CCM2 protein interactions in cell signaling: implications for cerebral cavernous malformations pathogenesis.

Cerebral cavernous malformations (CCMs) are sporadically acquired or inherited vascular lesions of the central nervous system consisting of clusters of dilated thin-walled blood vessels that predispose individuals to seizures and stroke. Familial CCM is caused by mutations in KRIT1 (CCM1) or in malcavernin (CCM2), the murine ortholog of which was concurrently characterized as osmosensing scaffold for MEKK3 (OSM). The roles of the CCM proteins in the pathogenesis of the disorder remain largely unknown. Here, we use co-immunoprecipitation, fluorescence resonance energy transfer and subcellular localization strategies to show that the CCM1 gene product, KRIT1, interacts with the CCM2 gene product, malcavernin/OSM. Analogous to the established interactions of CCM1 and beta1 integrin with ICAP1, the CCM1/CCM2 association is dependent upon the phosphotyrosine binding (PTB) domain of CCM2. A familial CCM2 missense mutation abrogates the CCM1/CCM2 interaction, suggesting that loss of this interaction may be critical in CCM pathogenesis. CCM2 and ICAP1 bound to CCM1 via their respective PTB domains differentially influence the subcellular localization of CCM1. Furthermore, we expand upon the established involvement of CCM2 in the p38 mitogen-activated protein kinase signaling module by demonstrating that CCM1 associates with CCM2 and MEKK3 in a ternary complex. These data indicate that the genetic heterogeneity observed in familial CCM may reflect mutation of different molecular members of a coordinated signaling complex.

Authors
Zawistowski, JS; Stalheim, L; Uhlik, MT; Abell, AN; Ancrile, BB; Johnson, GL; Marchuk, DA
MLA Citation
Zawistowski, JS, Stalheim, L, Uhlik, MT, Abell, AN, Ancrile, BB, Johnson, GL, and Marchuk, DA. "CCM1 and CCM2 protein interactions in cell signaling: implications for cerebral cavernous malformations pathogenesis." Hum Mol Genet 14.17 (September 1, 2005): 2521-2531.
PMID
16037064
Source
pubmed
Published In
Human Molecular Genetics
Volume
14
Issue
17
Publish Date
2005
Start Page
2521
End Page
2531
DOI
10.1093/hmg/ddi256

Subcellular localization of spastin: implications for the pathogenesis of hereditary spastic paraplegia.

Hereditary spastic paraplegia (HSP) is a group of clinically and genetically heterogeneous diseases characterized by neuronal degeneration that is maximal at the distal ends of the longest axons of the central nervous system. The most common cause of autosomal dominant HSP is mutation of a novel gene encoding spastin, a protein whose function is still being elucidated. One clue concerning spastin function is its intracellular localization. Here, we describe a novel anti-spastin antiserum designed to a unique epitope contained within all splicing isoforms. The antiserum exhibits specific immunostaining of recombinant spastin in intact, fixed cells. Using this reagent, we find that endogenous spastin is located at the centrosome in a variety of cell types at all points in the cell cycle. This localization is resistant to microtubule disruption, suggesting that spastin may be an integral centrosomal protein. In addition to the centrosome, spastin also localizes at discrete focal regions along the axons of primary cultured neurons. These data lend additional support to the emerging hypothesis that spastin plays a role in microtubule dynamics, with a crucial role in microtubule organization.

Authors
Svenson, IK; Kloos, MT; Jacon, A; Gallione, C; Horton, AC; Pericak-Vance, MA; Ehlers, MD; Marchuk, DA
MLA Citation
Svenson, IK, Kloos, MT, Jacon, A, Gallione, C, Horton, AC, Pericak-Vance, MA, Ehlers, MD, and Marchuk, DA. "Subcellular localization of spastin: implications for the pathogenesis of hereditary spastic paraplegia." Neurogenetics 6.3 (September 2005): 135-141.
PMID
15891913
Source
pubmed
Published In
Neurogenetics
Volume
6
Issue
3
Publish Date
2005
Start Page
135
End Page
141
DOI
10.1007/s10048-005-0219-2

Genetics of cerebral cavernous malformations.

The past few years have seen rapid advances in our understanding of the genetics and molecular biology of cerebral cavernous malformations (CCM). This article summarizes the recent cloning of the CCM1, CCM2, and CCM3 genes, which are responsible for autosomal dominant CCM, and also describes current hypotheses for their roles in integrin and p38 mitogen-activated protein kinase- mediated regulation of angiogenesis. A mouse model of CCM has been generated by mutation of the Ccm1 gene, and it indicates a role for that protein in arterial development. Future studies will probably focus on integration of data from each of the three CCM genes into a single model of the pathogenesis of cavernous malformation.

Authors
Plummer, NW; Zawistowski, JS; Marchuk, DA
MLA Citation
Plummer, NW, Zawistowski, JS, and Marchuk, DA. "Genetics of cerebral cavernous malformations." Curr Neurol Neurosci Rep 5.5 (September 2005): 391-396. (Review)
PMID
16131422
Source
pubmed
Published In
Current Neurology and Neuroscience Reports
Volume
5
Issue
5
Publish Date
2005
Start Page
391
End Page
396

QTL mapping in a mouse model of cardiomyopathy reveals an ancestral modifier allele affecting heart function and survival.

The progression from myocardial hypertrophy to heart failure is a complex process, involving genetic and environmental factors. Elucidating the genetic components contributing to heart failure has been difficult, largely because of the heterogeneity of human populations. We have employed a strategy to map genetic loci that modify the heart failure phenotype in a transgenic mouse model of cardiomyopathy caused by cardiac-specific overexpression of calsequestrin. Strain-specific differences in both cardiac function and survival are observed when the transgene is moved into different inbred mouse strains. We have previously reported linkage results from mapping in reciprocal backcrosses between C57/BL6 (BL6) and DBA/2J (DBA) and a backcross between DBA/AKR and AKR. Here we report the results of a genome-wide linkage scan in the reciprocal backcross between DBA/AKR and DBA. We identified one novel locus on Chromosome (Chr) 18 that affects heart function and a second on Chr 3 that shows significant linkage to both survival and heart function. Intriguingly, the Chr 3 allele of AKR shows a susceptibility effect on phenotype, whereas the overall effect of the AKR genetic background is protective. The Chr 3 locus also completely overlaps the Hrtfm2 locus, which was previously mapped in crosses between DBA and BL6. Mapping the same QTL in two different crosses allowed us to use ancestral haplotypes to narrow the candidate gene interval from 9 to 2 Mb. Identification of the genes at these QTLs in the mouse will provide novel candidate genes that can be evaluated for their role in human heart failure.

Authors
Wheeler, FC; Fernandez, L; Carlson, KM; Wolf, MJ; Rockman, HA; Marchuk, DA
MLA Citation
Wheeler, FC, Fernandez, L, Carlson, KM, Wolf, MJ, Rockman, HA, and Marchuk, DA. "QTL mapping in a mouse model of cardiomyopathy reveals an ancestral modifier allele affecting heart function and survival." Mamm Genome 16.6 (June 2005): 414-423.
PMID
16075368
Source
pubmed
Published In
Mammalian Genome
Volume
16
Issue
6
Publish Date
2005
Start Page
414
End Page
423
DOI
10.1007/s00335-005-2468-7

Influence of serotonin transporter promoter region polymorphisms on hippocampal volumes in late-life depression.

CONTEXT: Polymorphisms in the promoter region of the serotonin transporter gene (5-HTTLPR) influence transcription and may play a role in the pathogenesis and course of depression. Recent research demonstrates that specific polymorphisms may be associated with differences in hippocampal volumes in subjects with depression. OBJECTIVE: To examine associations between 5-HTTLPR genotype and hippocampal volumes in elderly control subjects and elderly subjects classified as having early or late onset of depression. DESIGN: Cohort study examining baseline characteristics. PARTICIPANTS: Subjects were community dwelling and 60 years or older. Using a definition of early-onset depression as depression first occurring at 50 years or younger, we examined 72 subjects with early-onset depression, 63 subjects with late-onset depression, and 83 healthy control subjects. MAIN OUTCOME MEASURES: All subjects underwent genotyping for the 5-HTTLPR and underwent brain magnetic resonance imaging. Analyses of hippocampal volumes were controlled for total cerebral volume, age, and sex. RESULTS: The interaction between diagnosis and 5-HTTLPR genotype was statistically significant for the right hippocampus (P = .04). Subjects with late-onset depression who were homozygous for the long (L) allele (L /L genotype) had significantly smaller right hippocampal volumes than did L /L subjects with early-onset depression (P = .046) or L /L control subjects (P = .01). Post hoc analyses showed that later age of depression onset was associated with smaller hippocampal volumes in subjects with the L /L genotype, but earlier age of onset was associated with smaller hippocampal volumes in subjects who were homozygous for the short (S) allele (S/S genotype). CONCLUSIONS: Subjects with late-onset depression who were homozygous for the L allele exhibited smaller hippocampal volumes than other groups. Genotype also mediated the effect of age of onset on hippocampal volumes. Our findings differ from previous work; however, we examined an older and larger cohort of subjects than previous studies. Possible explanations for these findings include interactions between the serotonergic system and neurotrophic factors or cortisol response to stresses, each of which may affect hippocampal volumes.

Authors
Taylor, WD; Steffens, DC; Payne, ME; MacFall, JR; Marchuk, DA; Svenson, IK; Krishnan, KRR
MLA Citation
Taylor, WD, Steffens, DC, Payne, ME, MacFall, JR, Marchuk, DA, Svenson, IK, and Krishnan, KRR. "Influence of serotonin transporter promoter region polymorphisms on hippocampal volumes in late-life depression." Arch Gen Psychiatry 62.5 (May 2005): 537-544.
PMID
15867107
Source
pubmed
Published In
Archives of General Psychiatry
Volume
62
Issue
5
Publish Date
2005
Start Page
537
End Page
544
DOI
10.1001/archpsyc.62.5.537

Human retroviral gag- and gag-pol-like proteins interact with the transforming growth factor-beta receptor activin receptor-like kinase 1.

Mutations in activin receptor-like kinase 1 (ALK1), a transforming growth factor (TGF)-beta type I receptor, lead to the vascular disorder hereditary hemorrhagic telangiectasia caused by abnormal vascular remodeling. The underlying molecular cause of this disease is not well understood. Identifying binding partners for ALK1 will help to understand its cellular function. Using the two-hybrid system, we identified an ALK1-binding protein encoded by an ancient retroviral/retrotransposon element integrated as a single copy gene known as PEG10 on human chromosome 7q21. PEG10 contains two overlapping reading frames from which two proteins, PEG10-RF1 and PEG10-RF1/2, are translated by a typical retroviral -1 ribosomal frameshift mechanism. Reverse transcription-PCR and Northern blot analysis showed a broad range of PEG10 expression in different tissues and cell types, i.e. human placenta, brain, kidney, endothelial cells, lymphoblasts, and HepG2 and HEK293 cells. However, endogenous PEG10-RF1 and PEG10-RF1/2 proteins were only detected in HepG2 and HEK293 cells. PEG10-RF1, which is the major PEG10 protein product, represents a gag-like protein, and PEG10-RF1/2 represents a gag-pol-like protein. PEG10-RF1 also interacts with different members of TGF-beta superfamily type I and II receptors. PEG10-RF1 binding to ALK1 is mediated by a 200-amino acid domain with no recognized motif. PEG10-RF1 inhibits ALK1 as well as ALK5 signaling. Co-expression of ALK1 and PEG10-RF1 in different cell types induced morphological changes reminiscent of neuronal cells or sprouting cells. This is the first report of a human retroviral-like protein interacting with members of the TGF-beta receptor family.

Authors
Lux, A; Beil, C; Majety, M; Barron, S; Gallione, CJ; Kuhn, H-M; Berg, JN; Kioschis, P; Marchuk, DA; Hafner, M
MLA Citation
Lux, A, Beil, C, Majety, M, Barron, S, Gallione, CJ, Kuhn, H-M, Berg, JN, Kioschis, P, Marchuk, DA, and Hafner, M. "Human retroviral gag- and gag-pol-like proteins interact with the transforming growth factor-beta receptor activin receptor-like kinase 1." J Biol Chem 280.9 (March 4, 2005): 8482-8493.
PMID
15611116
Source
pubmed
Published In
The Journal of biological chemistry
Volume
280
Issue
9
Publish Date
2005
Start Page
8482
End Page
8493
DOI
10.1074/jbc.M409197200

Functional analysis of a mutant form of the receptor tyrosine kinase Tie2 causing venous malformations.

Tie2 is expressed predominantly in endothelial cells and is required for blood vessel formation and maintenance. A missense mutation resulting in an R to W substitution in the kinase domain of Tie2 co-segregates with an autosomal dominantly inherited form of vascular dysmorphogenesis, venous malformation (VM). The mechanism by which this activating mutation leads to vessel dysmorphogenesis in VM is not known. Here we examined Tie2 activation status in VM and found activated receptor in lesional and non-lesional vessels. To gain insight into functional effects of VM mutant Tie2, wild-type and R849W mutant receptor were expressed in cultured human venous endothelial cells. Mutant Tie2 was constitutively phosphorylated in endothelial cells in vivo and caused a marked suppression of apoptosis. The anti-apoptotic kinase Akt was constitutively activated in cells expressing mutant receptor. Dominant-negative Akt inhibited the pro-survival activity of mutant Tie2. Migration of smooth muscle cells induced by conditioned medium from cells expressing mutant receptor was similar to that from cells expressing wild-type receptor. These data suggest that a primary effect of R849W Tie2 in VM is to allow survival of mural cell poor vessels via ligand-independent Tie2 activation of Akt and endothelial survival, rather than to directly induce formation of dysmorphogenic vessels.

Authors
Morris, PN; Dunmore, BJ; Tadros, A; Marchuk, DA; Darland, DC; D'Amore, PA; Brindle, NPJ
MLA Citation
Morris, PN, Dunmore, BJ, Tadros, A, Marchuk, DA, Darland, DC, D'Amore, PA, and Brindle, NPJ. "Functional analysis of a mutant form of the receptor tyrosine kinase Tie2 causing venous malformations." J Mol Med (Berl) 83.1 (January 2005): 58-63.
PMID
15526080
Source
pubmed
Published In
Journal of Molecular Medicine
Volume
83
Issue
1
Publish Date
2005
Start Page
58
End Page
63
DOI
10.1007/s00109-004-0601-9

Polymorphisms in transforming growth factor-β-related genes ALK1 and ENG are associated with sporadic brain arteriovenous malformations

Background and Purpose - Mutations in endoglin (ENG) and activin-like kinase (ALK1) cause hereditary hemorrhagic telangiectasias, disorders characterized by pulmonary and brain arteriovenous malformations (BAVMs). We investigated whether polymorphisms in these genes are also associated with sporadic BAVM. Methods - A total of 177 sporadic BAVM patients and 129 controls (all subjects white) were genotyped for 2 variants in ALK1 and 7 variants in ENG. Results - The ALK1 IVS3-35A>G polymorphism was associated with BAVM: (AnyA [AA+AG] genotype: odds ratio, 2.47; 95% CI, 1.38 to 4.44; P=0.002). Two ENG polymorphisms, ENG - 1742A>G and ENG 207G>A, showed a trend toward association with BAVM that did not reach statistical significance. Conclusions - A common polymorphism in ALK1 is associated with sporadic BAVM, suggesting that genetic variation in genes mutated in familial BAVM syndromes may play a role in sporadic BAVMs. © 2005 American Heart Association, Inc.

Authors
Pawlikowska, L; Tran, MN; Achrol, AS; Ha, C; Burchard, E; Choudhry, S; Zaroff, J; Lawton, MT; Castro, R; McCulloch, CE; Marchuk, D; Kwok, P-Y; Young, WL
MLA Citation
Pawlikowska, L, Tran, MN, Achrol, AS, Ha, C, Burchard, E, Choudhry, S, Zaroff, J, Lawton, MT, Castro, R, McCulloch, CE, Marchuk, D, Kwok, P-Y, and Young, WL. "Polymorphisms in transforming growth factor-β-related genes ALK1 and ENG are associated with sporadic brain arteriovenous malformations." Stroke 36.10 (2005): 2278-2280.
PMID
16179574
Source
scival
Published In
Stroke
Volume
36
Issue
10
Publish Date
2005
Start Page
2278
End Page
2280
DOI
10.1161/01.STR.0000182253.91167.fa

Loss of p53 sensitizes mice with a mutation in Ccm1 (KRIT1) to development of cerebral vascular malformations.

Cerebral cavernous malformations (CCM) consist of clusters of abnormally dilated blood vessels. Hemorrhaging of these lesions can cause seizures and lethal stroke. Three loci are associated with autosomal dominant CCM, and the causative genes have been identified for CCM1 and CCM2. We have generated mice with a targeted mutation of the Ccm1 gene, but an initial survey of 20 heterozygous mice failed to detect any cavernous malformations. To test the hypothesis that growth of cavernous malformations depends on somatic loss of heterozygosity at the Ccm1 locus, we bred animals that were heterozygous for the Ccm1 mutation and homozygous for loss of the tumor suppressor Trp53 (p53), which has been shown to increase the rate of somatic mutation. We observed vascular lesions in the brains of 55% of the double-mutant animals but none in littermates with other genotypes. Although the genetic evidence suggested somatic mutation of the wild-type Ccm1 allele, we were unable to demonstrate loss of heterozygosity by molecular methods. An alternative explanation is that p53 plays a direct role in formation of the vascular malformations. The striking similarity of the human and mouse lesions indicates that the Ccm1(+/-) Trp53(-/-) mice are an appropriate animal model of CCM.

Authors
Plummer, NW; Gallione, CJ; Srinivasan, S; Zawistowski, JS; Louis, DN; Marchuk, DA
MLA Citation
Plummer, NW, Gallione, CJ, Srinivasan, S, Zawistowski, JS, Louis, DN, and Marchuk, DA. "Loss of p53 sensitizes mice with a mutation in Ccm1 (KRIT1) to development of cerebral vascular malformations." Am J Pathol 165.5 (November 2004): 1509-1518.
PMID
15509522
Source
pubmed
Published In
The American journal of pathology
Volume
165
Issue
5
Publish Date
2004
Start Page
1509
End Page
1518
DOI
10.1016/S0002-9440(10)63409-8

Polymorphisms in genes involved in inflammatory and angiogenic pathways and the risk of hemorrhagic presentation of brain arteriovenous malformations.

BACKGROUND AND PURPOSE: Accurate estimates of intracranial hemorrhage (ICH) risk in patients harboring brain arteriovenous malformation (BAVM) are needed to evaluate interventional strategies and to help guide clinical management. Identification of genetic polymorphisms associated with ICH would facilitate risk stratification in BAVM patients. METHODS: We identified patients with BAVM and documented clinical presentation, demographic data, venous drainage pattern, and BAVM size. Patients were genotyped for 5 polymorphisms in 3 inflammatory cytokine genes, and 9 polymorphisms in 5 angiogenesis-related genes. Association of genotype with risk of hemorrhagic BAVM presentation was evaluated using logistic regression analysis. RESULTS: We genotyped 180 patients with BAVM (53% female, 57% white, mean age at diagnosis 35+/-17 years, 41% presenting with ICH). BAVM patients homozygous for the interleukin 6 (IL6)-174G allele had a greater risk of ICH presentation (OR, 2.62, P=0.003) than IL6-174C carriers. In a multivariate logistic regression model, IL6-174G>C genotype, small BAVM size, and exclusively deep venous drainage were independent predictors of ICH presentation. A similar univariate trend was noted for the TNFalpha-308 GG genotype (P=0.055). The other polymorphisms genotyped were not associated with ICH. CONCLUSIONS: A polymorphism in the inflammatory cytokine IL6, but not polymorphisms in angiogenesis-related genes, was associated with ICH presentation of BAVM. Further studies are needed to define the role of inflammatory cytokines in the pathogenesis of BAVM hemorrhage.

Authors
Pawlikowska, L; Tran, MN; Achrol, AS; McCulloch, CE; Ha, C; Lind, DL; Hashimoto, T; Zaroff, J; Lawton, MT; Marchuk, DA; Kwok, P-Y; Young, WL; UCSF BAVM Study Project,
MLA Citation
Pawlikowska, L, Tran, MN, Achrol, AS, McCulloch, CE, Ha, C, Lind, DL, Hashimoto, T, Zaroff, J, Lawton, MT, Marchuk, DA, Kwok, P-Y, Young, WL, and UCSF BAVM Study Project, . "Polymorphisms in genes involved in inflammatory and angiogenic pathways and the risk of hemorrhagic presentation of brain arteriovenous malformations." Stroke 35.10 (October 2004): 2294-2300.
PMID
15331795
Source
pubmed
Published In
Stroke
Volume
35
Issue
10
Publish Date
2004
Start Page
2294
End Page
2300
DOI
10.1161/01.STR.0000141932.44613.b1

Giant infiltrative cavernous malformation: clinical presentation, intervention, and genetic analysis: case report.

OBJECTIVE AND IMPORTANCE: Cavernous malformations can present in children with a sporadic course of repeated hemorrhage and enlargement, but they are rarely aggressive, infiltrative, or multilobar. We present the case of a young boy with a complex cavernous malformation that evolved during the course of a decade to encompass the majority of his right cerebral hemisphere. CLINICAL PRESENTATION: A 16-month-old boy presented with seizures, and radiographic studies demonstrated a large cavernous malformation in his right frontal pole. During the next 10 years, his seizures became intractable, and he developed progressive left hand weakness and atrophy. His malformation infiltrated his entire right frontal lobe as well as portions of his right parietal lobe, temporal lobe, and deep gray matter structures. INTERVENTION: The patient underwent right hemicraniotomy and near total resection of the lesion. Pathological analysis revealed dilated, thin-walled vessels separated by small amounts of intervening astrogliotic brain consistent with cavernous malformation. The patient recovered to his baseline neurological condition and has had no seizure or hemorrhage since his operation. Genetic testing did not reveal mutations in either the CCM1 (KRIT1) or CCM2 (malcavernin) genes. CONCLUSION: This case may represent an atypical variant of cavernous malformation best termed giant infiltrative cavernous malformation. Despite its unusual size, multilobar location, and aggressive infiltration, it can be managed effectively with standard surgical resection.

Authors
Lawton, MT; Vates, GE; Quinones-Hinojosa, A; McDonald, WC; Marchuk, DA; Young, WL
MLA Citation
Lawton, MT, Vates, GE, Quinones-Hinojosa, A, McDonald, WC, Marchuk, DA, and Young, WL. "Giant infiltrative cavernous malformation: clinical presentation, intervention, and genetic analysis: case report." Neurosurgery 55.4 (October 2004): 979-980.
PMID
15934180
Source
pubmed
Published In
Neurosurgery
Volume
55
Issue
4
Publish Date
2004
Start Page
979
End Page
980

Intragenic modifiers of hereditary spastic paraplegia due to spastin gene mutations.

Hereditary spastic paraplegia (HSP) is a genetically heterogeneous neurodegenerative disease characterized by wide variability in phenotypic expression, both within and among families. The most-common cause of autosomal dominant HSP is mutation of the gene encoding spastin, a protein of uncertain function. We report the existence of intragenic polymorphisms of spastin that modify the HSP phenotype. One (S44L) is a previously described recessively acting allele and the second is a novel allele affecting the adjacent amino acid residue (P45Q). In 4 HSP families in which either L44 or Q45 segregates independently of a missense or splicing mutation in the AAA domain of spastin, L44 and Q45 are each associated with a striking decrease in age at onset in the presence of the AAA domain mutations. Using a bioinformatics approach, we found that the highly conserved S44 is predicted to be phosphorylated by a number of family members of the proline-directed serine/threonine cyclin-dependent kinases (Cdks). Cdk1 and Cdk5 showed no kinase activity toward synthetic spastin peptide in an in vitro kinase assay, suggesting that this serine residue may be phosphorylated by a different Cdk. Our identification of S44L and P45Q as modifiers of the HSP phenotype suggests a role for spastin phosphorylation by Cdks in the neurodegeneration of the most-common form of HSP.

Authors
Svenson, IK; Kloos, MT; Gaskell, PC; Nance, MA; Garbern, JY; Hisanaga, S-I; Pericak-Vance, MA; Ashley-Koch, AE; Marchuk, DA
MLA Citation
Svenson, IK, Kloos, MT, Gaskell, PC, Nance, MA, Garbern, JY, Hisanaga, S-I, Pericak-Vance, MA, Ashley-Koch, AE, and Marchuk, DA. "Intragenic modifiers of hereditary spastic paraplegia due to spastin gene mutations." Neurogenetics 5.3 (September 2004): 157-164.
PMID
15248095
Source
pubmed
Published In
Neurogenetics
Volume
5
Issue
3
Publish Date
2004
Start Page
157
End Page
164
DOI
10.1007/s10048-004-0186-z

CCM2 mutations account for 13% of cases in a large collection of kindreds with hereditary cavernous malformations.

Authors
Verlaan, DJ; Laurent, SB; Rochefort, DL; Liquori, CL; Marchuk, DA; Siegel, AM; Rouleau, GA
MLA Citation
Verlaan, DJ, Laurent, SB, Rochefort, DL, Liquori, CL, Marchuk, DA, Siegel, AM, and Rouleau, GA. "CCM2 mutations account for 13% of cases in a large collection of kindreds with hereditary cavernous malformations." Ann Neurol 55.5 (May 2004): 757-758. (Letter)
PMID
15122722
Source
pubmed
Published In
Annals of Neurology
Volume
55
Issue
5
Publish Date
2004
Start Page
757
End Page
758
DOI
10.1002/ana.20112

A combined syndrome of juvenile polyposis and hereditary haemorrhagic telangiectasia associated with mutations in MADH4 (SMAD4).

BACKGROUND: Juvenile polyposis and hereditary haemorrhagic telangiectasia are autosomal dominant disorders with distinct and non-overlapping clinical features. The former, an inherited gastrointestinal malignancy predisposition, is caused by mutations in MADH4 (encoding SMAD4) or BMPR1A, and the latter is a vascular malformation disorder caused by mutations in ENG (endoglin) or ACVRL1 (ALK1). All four genes encode proteins involved in the transforming-growth-factor-beta signalling pathway. Although there are reports of patients and families with phenotypes of both disorders combined, the genetic aetiology of this association is unknown. METHODS: Blood samples were collected from seven unrelated families segregating both phenotypes. DNA from the proband of each family was sequenced for the ACVRL1, ENG, and MADH4 genes. Mutations were examined for familial cosegregation with phenotype and presence or absence in population controls. Findings No patient had mutations in the ENG or ACVRL1 genes; all had MADH4 mutations. Three cases of de-novo MADH4 mutations were found. In one, the mutation was passed on to a similarly affected child. Each mutation cosegregated with the syndromic phenotype in other affected family members. INTERPRETATION: Mutations in MADH4 can cause a syndrome consisting of both juvenile polyposis and hereditary haemorrhagic telangiectasia phenotypes. Since patients with these disorders are generally ascertained through distinct medical specialties, genetic testing is recommended for patients presenting with either phenotype to identify those at risk of this syndrome. Patients with juvenile polyposis who have an MADH4 mutation should be screened for the vascular lesions associated with hereditary haemorrhagic telangiectasia, especially occult arteriovenous malformations in visceral organs that may otherwise present suddenly with serious medical consequences.

Authors
Gallione, CJ; Repetto, GM; Legius, E; Rustgi, AK; Schelley, SL; Tejpar, S; Mitchell, G; Drouin, E; Westermann, CJJ; Marchuk, DA
MLA Citation
Gallione, CJ, Repetto, GM, Legius, E, Rustgi, AK, Schelley, SL, Tejpar, S, Mitchell, G, Drouin, E, Westermann, CJJ, and Marchuk, DA. "A combined syndrome of juvenile polyposis and hereditary haemorrhagic telangiectasia associated with mutations in MADH4 (SMAD4)." Lancet 363.9412 (March 13, 2004): 852-859.
PMID
15031030
Source
pubmed
Published In
The Lancet
Volume
363
Issue
9412
Publish Date
2004
Start Page
852
End Page
859
DOI
10.1016/S0140-6736(04)15732-2

Primary pulmonary hypertension in families with hereditary haemorrhagic telangiectasia.

Primary pulmonary hypertension (PPH) is a rare but severe and progressive disease characterised by obstructive lesions of small pulmonary arteries. Patients with PPH often have mutations in the bone morphogenetic protein receptor type II (BMPR2) gene, whereas some carry mutations in the activin receptor-like kinase 1 (ALK-1) gene, generally associated with hereditary haemorrhagic telangiectasia (HHT) type 2, a vascular dysplasia affecting multiple organs. The aim of this study was to determine whether members of families with PPH and confirmed or probable HHT had ALK-1 mutations. ALK-1 and BMPR2 mutation analysis was performed on deoxyribonucleic acid from affected members of four families with PPH and confirmed or suspected HHT. ALK-1 mutations were identified in all four families and three novel mutations found in exon 10, leading to truncated proteins. In the fourth family, a missense mutation, previously reported in four independent HHT families, was detected in exon 8. Analysis of the BMPR2 gene revealed no exonic mutations in the probands with both PPH and HHT. The present data bring to 10 the number of reported families with primary pulmonary hypertension and hereditary haemorrhagic telangiectasia type 2, representing 16% of the 61 families with known activin receptor-like kinase 1 mutations. Such mutations might predispose to primary pulmonary hypertension, and specialists should be aware of the potential link between these two disorders.

Authors
Abdalla, SA; Gallione, CJ; Barst, RJ; Horn, EM; Knowles, JA; Marchuk, DA; Letarte, M; Morse, JH
MLA Citation
Abdalla, SA, Gallione, CJ, Barst, RJ, Horn, EM, Knowles, JA, Marchuk, DA, Letarte, M, and Morse, JH. "Primary pulmonary hypertension in families with hereditary haemorrhagic telangiectasia." Eur Respir J 23.3 (March 2004): 373-377.
PMID
15065824
Source
pubmed
Published In
The European respiratory journal : official journal of the European Society for Clinical Respiratory Physiology
Volume
23
Issue
3
Publish Date
2004
Start Page
373
End Page
377

Ccm1 is required for arterial morphogenesis: implications for the etiology of human cavernous malformations.

Hemorrhagic stroke is a significant cause of morbidity and mortality in children, and is frequently associated with intracranial vascular malformations. One prevalent form of these vascular malformations, cerebral cavernous malformation, is characterized by thin-walled vascular cavities that hemorrhage and has been linked to loss-of-function mutations in CCM1. The neural and epithelial expression of CCM1 in adulthood suggests that cavernous malformations may be the result of primary neural defects. In this study, we generated mice lacking Ccm1 and demonstrate that Ccm1 is ubiquitously expressed early in embryogenesis and is essential for vascular development. Homozygous mutant embryos die in mid-gestation and the first detectable defects are exclusively vascular in nature. The precursor vessels of the brain become dilated starting at E8.5, reminiscent of the intracranial vascular defects observed in the human disease. In addition, there is marked enlargement and increased endothelial proliferation of the caudal dorsal aorta, as well as variable narrowing of the branchial arch arteries and proximal dorsal aorta. These vascular defects are not secondary to primary neural defects, as neural morphology and marker expression are normal even subsequent to the onset of vascular pathology. The defects in the vascular structure of embryos lacking Ccm1 are associated with early downregulation of artery-specific markers, including the Efnb2- and Notch-related genes. Finally, consistent with the murine data, we found that there is an analogous reduction in Notch gene expression in arterioles from humans with mutations in CCM1. Our studies suggest that cavernous malformations result from primary vascular rather than neural defects.

Authors
Whitehead, KJ; Plummer, NW; Adams, JA; Marchuk, DA; Li, DY
MLA Citation
Whitehead, KJ, Plummer, NW, Adams, JA, Marchuk, DA, and Li, DY. "Ccm1 is required for arterial morphogenesis: implications for the etiology of human cavernous malformations." Development 131.6 (March 2004): 1437-1448.
PMID
14993192
Source
pubmed
Published In
Development (Cambridge)
Volume
131
Issue
6
Publish Date
2004
Start Page
1437
End Page
1448
DOI
10.1242/dev.01036

Modifier locus on mouse chromosome 3 for renal vascular pathology in AT1A receptor-deficiency.

We previously showed that the phenotype of mice with targeted disruption of the gene encoding the AT1A receptor (Agtr1a), the major murine AT1 receptor isoform, is strongly influenced by recessive genetic modifiers derived from the C57BL/6 or 129 inbred strains. To further evaluate the genetic modifiers on the C57BL/6 background, we performed backcrosses between F1(C57BL/6x129) and C57BL/6 Agtr1a-/- mice and analyzed the progeny, focusing on the development of structural lesions in the renal vasculature. In affected animals, these lesions are characterized by medial thickening of small arteries and arterioles in the kidney that are reminiscent of vascular lesions in patients with nephrosclerosis. Among 180 consecutive progeny, 170 (94%) survived to completion of the study. On masked pathological examination at age 8 months, 86 had intermediate to severe vascular lesions whereas 84 had no detectable lesions. Based on a hypothetical model of a single recessive modifier locus arising from the C57BL/6 background, the observed proportion of affected animals among the backcross progeny was not statistically different from that predicted by chi2 analysis (51% versus 50%; P=0.88). We next performed genomic microsatellite analysis in a subset of 121 backcross progeny using a panel of markers spanning approximately 15 cM intervals across the mouse genome. By 2-point analysis, we found a region spanning 5 cM on chromosome 3, with significant linkage to the development of renal vascular lesions (LOD score: 3.3 to 3.8).

Authors
Le, TH; Fogo, AB; Salzler, HR; Vinogradova, T; Oliverio, MI; Marchuk, DA; Coffman, TM
MLA Citation
Le, TH, Fogo, AB, Salzler, HR, Vinogradova, T, Oliverio, MI, Marchuk, DA, and Coffman, TM. "Modifier locus on mouse chromosome 3 for renal vascular pathology in AT1A receptor-deficiency." Hypertension 43.2 (February 2004): 445-451.
PMID
14718357
Source
pubmed
Published In
Hypertension
Volume
43
Issue
2
Publish Date
2004
Start Page
445
End Page
451
DOI
10.1161/01.HYP.0000112423.28987.00

Gene microarray analysis of human brain arteriovenous malformations.

OBJECTIVE: Human brain arteriovenous malformations (BAVMs) display abnormal expression of various angiogenesis-related genes and their products. We examined gene expression patterns in BAVMs by the gene microarray technique. METHODS: We analyzed BAVM and control brain samples obtained by temporal lobectomy for medically intractable seizure by Affymetrix Human Gene Set U95Av2 (Affymetrix, Inc., Santa Clara, CA). The gene microarray data were compared with new and previously published data that used conventional molecular biology techniques. RESULTS: We analyzed six BAVM and five control brain samples. From 12,625 gene probes assayed, 1781 gene probes showed differential expression between BAVMs and controls. BAVM samples had a gene expression pattern that was distinct from those of control brain samples. Increased messenger ribonucleic acid expression of vascular endothelial growth factor A was accompanied by increased expression of its protein product. A majority of the gene data was in agreement with previously published data. The gene microarray data generated a new testable hypothesis regarding integrin, and we found increased expression of integrin alphavbeta3 protein in BAVMs. CONCLUSION: The gene expression pattern of BAVMs was distinct from those of control brain samples. We verified the gene microarray data by demonstrating that increased gene expression levels for angiogenesis-related molecules were accompanied by increased levels of their protein product expression. The gene microarray technique may be a useful tool to study multiple pathways simultaneously in BAVM specimens.

Authors
Hashimoto, T; Lawton, MT; Wen, G; Yang, G-Y; Chaly, T; Stewart, CL; Dressman, HK; Barbaro, NM; Marchuk, DA; Young, WL
MLA Citation
Hashimoto, T, Lawton, MT, Wen, G, Yang, G-Y, Chaly, T, Stewart, CL, Dressman, HK, Barbaro, NM, Marchuk, DA, and Young, WL. "Gene microarray analysis of human brain arteriovenous malformations." Neurosurgery 54.2 (February 2004): 410-423.
PMID
14744289
Source
pubmed
Published In
Neurosurgery
Volume
54
Issue
2
Publish Date
2004
Start Page
410
End Page
423

Vascular endothelial growth factor induces abnormal microvasculature in the endoglin heterozygous mouse brain.

Hereditary hemorrhagic telangiectasia (HHT), associated with brain arteriovenous malformations, is caused by a loss of function mutation in either the endoglin (HHT1) or activin receptor-like kinase 1 gene (ALK-1, HHT2). Endoglin heterozygous (Eng+/-)mice have been proposed as a disease model. To better understand the role of endoglin in vascular malformation development, we examined the effect of vascular endothelial growth factor (VEGF) hyperstimulation on microvessels in adult endoglin heterozygous (Eng+/-) mice using an adenoviral vector to deliver recombinant human VEGF165 cDNA (AdhVEGF) into basal ganglia. VEGF expression was increased in AdhVEGF mice compared with the AdlacZ and saline group (P < 0.05) and localized to multiple cell types (neurons, astrocytes, endothelial cells, and smooth muscle cells) by double-labeled immunostaining. VEGF overexpression increased microvessel count for up to 4 weeks in both the Eng+/+ and Eng+/- groups (Eng+/+ 185 +/- 14 vs. Eng+/- 201 +/- 10 microvessels/mm2). Confocal microscopic examination revealed grossly abnormal microvessels in eight of nine Eng+/- mouse brains compared with zero of nine in Eng+/+ mice (P < 0.05). Abnormal microvessels featured enlargement, clustering, twist, or spirals. VEGF receptor Flk-1 and TGF-beta receptor 1 (T beta R1) expression were reduced in the Eng+/- mouse brain compared with control. Excessive VEGF stimulation may play a pivotal role in the initiation and development of brain vessel malformations in states of relative endoglin insufficiency in adulthood. These observations are relevant to our general understanding of the maintenance of vascular integrity.

Authors
Xu, B; Wu, YQ; Huey, M; Arthur, HM; Marchuk, DA; Hashimoto, T; Young, WL; Yang, G-Y
MLA Citation
Xu, B, Wu, YQ, Huey, M, Arthur, HM, Marchuk, DA, Hashimoto, T, Young, WL, and Yang, G-Y. "Vascular endothelial growth factor induces abnormal microvasculature in the endoglin heterozygous mouse brain." J Cereb Blood Flow Metab 24.2 (February 2004): 237-244.
PMID
14747750
Source
pubmed
Published In
Journal of Cerebral Blood Flow and Metabolism
Volume
24
Issue
2
Publish Date
2004
Start Page
237
End Page
244
DOI
10.1097/01.WCB.0000107730.66603.51

Multiple quantitative trait loci modify the heart failure phenotype in murine cardiomyopathy.

The variability in outcome of heart failure patients depends on a number of factors including differences in their genetic background. To identify novel genes that modify the human heart failure phenotype, we used a strategy of quantitative trait locus (QTL) mapping in an experimental mouse model of dilated cardiomyopathy induced by cardiac-specific overexpression of calsequestrin and characterized by a strong strain-specific variability in the phenotype. We identified two novel QTLs, Hrtfm3 (heart failure modifier 3) on chromosome (Chr) 4 and Hrtfm4 on Chr 18, significantly linked to survival with likelihood ratio statistics (LRS) of 19.9 and 23.6 respectively (corresponding to LOD scores of 4.3 and 5.1). Two other QTLs, Hrtfm5 on Chr 2 and Hrtfm6 on Chr 13, were significantly linked to cardiac function as measured by echocardiographic fractional shortening (LRS 22.1 and 15.2 respectively, LOD score 4.8 and 3.3) and left ventricular end-diastolic diameter (LRS 23.5 and 18.8, LOD score 5.1 and 4.1). Importantly, Hrtfm5 was not significantly linked to survival. A significant interaction was found between Hrtfm4 and two other QTLs (Hrtfm6 and a QTL near to the marker D19Mit88) for fractional shortening with a LRS of 34.6 and 26.5 respectively (LOD score 7.5 and 5.8). These data show that the effect of genetic background on murine heart failure is complex and result from the action of several loci that differentially modify the cardiac phenotype. The identification of these novel modifier genes will serve as strong candidates for the discovery of modifiers in human heart failure.

Authors
Le Corvoisier, P; Park, H-Y; Carlson, KM; Marchuk, DA; Rockman, HA
MLA Citation
Le Corvoisier, P, Park, H-Y, Carlson, KM, Marchuk, DA, and Rockman, HA. "Multiple quantitative trait loci modify the heart failure phenotype in murine cardiomyopathy." Hum Mol Genet 12.23 (December 1, 2003): 3097-3107.
PMID
14519689
Source
pubmed
Published In
Human Molecular Genetics
Volume
12
Issue
23
Publish Date
2003
Start Page
3097
End Page
3107
DOI
10.1093/hmg/ddg333

Mutations in a gene encoding a novel protein containing a phosphotyrosine-binding domain cause type 2 cerebral cavernous malformations.

Cerebral cavernous malformations (CCMs) are congenital vascular anomalies of the central nervous system that can result in hemorrhagic stroke, seizures, recurrent headaches, and focal neurologic deficits. Mutations in the gene KRIT1 are responsible for type 1 CCM (CCM1). We report that a novel gene, MGC4607, exhibits eight different mutations in nine families with type 2 CCM (CCM2). MGC4607, similar to the KRIT1 binding partner ICAP1alpha, encodes a protein with a phosphotyrosine-binding domain. This protein may be part of the complex pathway of integrin signaling that, when perturbed, causes abnormal vascular morphogenesis in the brain, leading to CCM formation.

Authors
Liquori, CL; Berg, MJ; Siegel, AM; Huang, E; Zawistowski, JS; Stoffer, T; Verlaan, D; Balogun, F; Hughes, L; Leedom, TP; Plummer, NW; Cannella, M; Maglione, V; Squitieri, F; Johnson, EW; Rouleau, GA; Ptacek, L; Marchuk, DA
MLA Citation
Liquori, CL, Berg, MJ, Siegel, AM, Huang, E, Zawistowski, JS, Stoffer, T, Verlaan, D, Balogun, F, Hughes, L, Leedom, TP, Plummer, NW, Cannella, M, Maglione, V, Squitieri, F, Johnson, EW, Rouleau, GA, Ptacek, L, and Marchuk, DA. "Mutations in a gene encoding a novel protein containing a phosphotyrosine-binding domain cause type 2 cerebral cavernous malformations." Am J Hum Genet 73.6 (December 2003): 1459-1464.
PMID
14624391
Source
pubmed
Published In
The American Journal of Human Genetics
Volume
73
Issue
6
Publish Date
2003
Start Page
1459
End Page
1464
DOI
10.1086/380314

Meta-analysis of age at onset in spastin-associated hereditary spastic paraplegia provides no evidence for a correlation with mutational class.

Authors
Yip, AG; Dürr, A; Marchuk, DA; Ashley-Koch, A; Hentati, A; Rubinsztein, DC; Reid, E
MLA Citation
Yip, AG, Dürr, A, Marchuk, DA, Ashley-Koch, A, Hentati, A, Rubinsztein, DC, and Reid, E. "Meta-analysis of age at onset in spastin-associated hereditary spastic paraplegia provides no evidence for a correlation with mutational class." J Med Genet 40.9 (September 2003): e106-.
PMID
12960222
Source
pubmed
Published In
Journal of medical genetics
Volume
40
Issue
9
Publish Date
2003
Start Page
e106

Vascular morphogenesis: tales of two syndromes.

Advances in our understanding of fundamental biological processes can be made by the analysis of defects manifested in inherited diseases. The genes responsible for these genetic syndromes often encode proteins that act at critical points of the pathways that control biological processes such as cell proliferation, cell-cell communication, cellular differentiation, and cell death. This approach has lead to the discovery of novel gene products and/or biochemical pathways involved in disease, genes that in turn play a fundamental role in normal biological processes. This forward genetic approach, focusing on Mendelian disorders of vascular anomalies, has been particularly fruitful for the study of genetic regulation of angiogenesis. This review summarizes the ongoing saga of two genetic syndromes involving disruption of normal vascular morphogenesis. Each inherited disorder involves the focal development of a distinct vascular anomaly. In hereditary hemorrhagic telangiectasia (HHT), the hallmark vascular lesion is termed an arteriovenous malformation, which involves the direct communication of an artery with a vein (arteriovenous shunt), without an intervening capillary bed. For cerebral cavernous malformations (CCM), the lesions are grossly-dilated, closely-packed, capillary-like sinusoidal chambers. The autosomal dominant mode of inheritance of each of these distinct syndromes suggested that the underlying genes might regulate critical aspects of vascular morphogenesis. Emerging but intriguing tales are being told by the genes (and their protein products) mutated in these disorders.

Authors
Marchuk, DA; Srinivasan, S; Squire, TL; Zawistowski, JS
MLA Citation
Marchuk, DA, Srinivasan, S, Squire, TL, and Zawistowski, JS. "Vascular morphogenesis: tales of two syndromes." Hum Mol Genet 12 Spec No 1 (April 1, 2003): R97-112. (Review)
PMID
12668602
Source
pubmed
Published In
Human Molecular Genetics
Volume
12 Spec No 1
Publish Date
2003
Start Page
R97
End Page
112

A mouse model for hereditary hemorrhagic telangiectasia (HHT) type 2.

Hereditary hemorrhagic telangiectasia (HHT) is an autosomal-dominant disorder characterized by the age-dependent development of focal arteriovenous malformations and telangiectases. HHT type 2 is caused by loss of function mutations in activin receptor-like kinase 1 (ACVRL1 or ALK1). However, the factors that initiate lesion formation and those that influence disease progression remain unknown. Because heterozygous mice contain the appropriate genotype for an animal model of this disorder, mice heterozygous for a loss-of-function mutation in Acvrl1 were carefully examined for an HHT-like phenotype. These mice developed age-dependent vascular lesions in the skin, extremities, oral cavity and in the internal organs (lung, liver, intestine, spleen and brain), similar to those seen in HHT patients. Major histopathological features of the lesions included thin-walled dilated vessels in close proximity to each other, hemorrhage and fibrosis. Similar to HHT patients, the mice also exhibited gastrointestinal bleeding, as evidenced by positive fecal occult blood tests. An Acvrl1(+/-) mouse with profound liver involvement also displayed a secondary cardiac phenotype, similar to that observed in human patients. The similarity of affected organs, age-dependent penetrance, histological similarity of the lesions and recapitulation of a secondary phenotype suggest that the Acvrl1(+/-) mice are an appropriate animal model for the identification of additional genetic and environmental factors that cause pathology in HHT type 2 patients. In addition, studies utilizing this animal model can yield valuable information on the role of ALK1 in maintenance of adult vascular architecture including arteriovenous identity.

Authors
Srinivasan, S; Hanes, MA; Dickens, T; Porteous, MEM; Oh, SP; Hale, LP; Marchuk, DA
MLA Citation
Srinivasan, S, Hanes, MA, Dickens, T, Porteous, MEM, Oh, SP, Hale, LP, and Marchuk, DA. "A mouse model for hereditary hemorrhagic telangiectasia (HHT) type 2." Hum Mol Genet 12.5 (March 1, 2003): 473-482.
PMID
12588795
Source
pubmed
Published In
Human Molecular Genetics
Volume
12
Issue
5
Publish Date
2003
Start Page
473
End Page
482

Impact of genetic polymorphisms on heart failure prognosis.

Recent progress in genomic applications have led to a better understanding of the relationship between genetic background and cardiovascular diseases such as heart failure. The broad variability in heart failure patient outcome is in part secondary to modifier genes, i.e. genes that are not involved in the genesis of a disease but modify the severity of the phenotypic expression once the disease has developed. The strategy most commonly used to identify modifier genes is based on association studies between the severity of the phenotype and the sequence variation(s) of selected candidate gene(s). Using this strategy, several polymorphisms of the beta 1 and beta 2-adrenergic receptors genes and the angiotensin converting enzyme gene have been correlated to the prognosis of patients with heart failure. Recently, we have applied an experimental strategy, known as genome mapping, for the identification of heart failure modifier genes. Genome mapping has previously been used with success to identify the genes involved in the development of both monogenic and multifactorial diseases. We have shown that the prognosis of heart failure mice, induced through calsequestrin overexpression, is linked to two Quantitative Trait Loci localized on chromosomes 2 and 3. Using both strategies (candidate gene and genome mapping) should allow us to identify a number of modifier genes that may provide a more rational approach to identify patients with the worst prognosis and to predict their response to therapy.

Authors
Le Corvoisier, P; Park, HY; Carlson, KM; Donahue, MP; Marchuk, DA; Rockman, HA
MLA Citation
Le Corvoisier, P, Park, HY, Carlson, KM, Donahue, MP, Marchuk, DA, and Rockman, HA. "Impact of genetic polymorphisms on heart failure prognosis." Arch Mal Coeur Vaiss 96.3 (March 2003): 197-206. (Review)
PMID
12722550
Source
pubmed
Published In
Archives des maladies du coeur et des vaisseaux
Volume
96
Issue
3
Publish Date
2003
Start Page
197
End Page
206

Serotonin-related gene polymorphisms and central nervous system serotonin function.

Central nervous system (CNS) serotonergic function affects a wide range of biological and behavioral functions affecting health and disease. Our objective in this study was to determine whether functional polymorphisms of the genes that encode for the serotonin transporter promoter (5HTTLPR) and monoamine oxidase A (MAOA-uVNTR) are associated with CNS serotonin turnover-indexed by cerebrospinal fluid levels of 5-hydroxyindoleacetic acid (5-HIAA)-in a community sample of healthy adults. Subjects were 165 community volunteers without current medical or psychiatric illness, stratified with respect to ethnicity, gender, and socioeconomic status who underwent inpatient evaluation in the General Clinical Research Center of a university medical center. A significant ethnicity x genotype interaction (P=0.008) indicated that, compared to the long/long and long/short genotypes, the 5HTTLPR short/short genotype was associated with higher CSF 5-HIAA levels in African Americans, but with lower levels in Caucasians. A gender x genotype interaction (P=0.04) indicated that 5HTTLPR short/short genotype was associated with higher 5-HIAA levels in women but with lower levels in men. MAOA-uVNTR 3.5 and 4 repeat alleles were associated with higher 5-HIAA (P=0.03) levels in men, but were unrelated to 5-HIAA levels in women. These findings suggest that effects of serotonin-related gene polymorphisms on CNS serotonergic function vary as a function of both ethnicity and gender. Further research will be required to determine the mechanism(s) underlying these differential effects. In the meanwhile, both ethnicity and gender should be taken into account in research evaluating effects of these and related polymorphisms on CNS serotonergic function, as well as the broad range of biological and behavioral functions that are regulated by CNS serotonergic function.

Authors
Williams, RB; Marchuk, DA; Gadde, KM; Barefoot, JC; Grichnik, K; Helms, MJ; Kuhn, CM; Lewis, JG; Schanberg, SM; Stafford-Smith, M; Suarez, EC; Clary, GL; Svenson, IK; Siegler, IC
MLA Citation
Williams, RB, Marchuk, DA, Gadde, KM, Barefoot, JC, Grichnik, K, Helms, MJ, Kuhn, CM, Lewis, JG, Schanberg, SM, Stafford-Smith, M, Suarez, EC, Clary, GL, Svenson, IK, and Siegler, IC. "Serotonin-related gene polymorphisms and central nervous system serotonin function." Neuropsychopharmacology 28.3 (March 2003): 533-541.
PMID
12629534
Source
pubmed
Published In
Neuropsychopharmacology (Nature)
Volume
28
Issue
3
Publish Date
2003
Start Page
533
End Page
541
DOI
10.1038/sj.npp.1300054

Associations among the NEO personality inventory, revised and the serotonin transporter gene-linked polymorphic region in elders: Effects of depression and gender

Objective: The short variant of the serotonin transporter gene-linked functional polymorphic region (5-HTTLPR) has been associated with personality traits related to anxiety, hostility, and depression. We attempted to replicate findings suggesting a positive relation between the short allele variant of 5-HTTLPR and Neuroticism, and a negative association between the short allele variant and Agreeableness. Methods: Participants in the present study were 103 geriatric depressed patients and 99 non-depressed age matched controls. Depression status and gender were examined as potential modifiers of the association between 5-HTTLPR and personality. Results: Neuroticism was associated with allele frequency such that individuals with the short variant of the allele (ss or sl, group S) were significantly lower on Neuroticism (P<0.04) compared with individuals with the long allele variant (group L), a pattern opposite to that of previous reports. The association did not vary by clinical group (depressed or controls) but was conditional on gender (P<0.01): the mean Neuroticism for males in group S was 48.2, whereas the mean Neuroticism for males in group L was 55.9; and the mean Neuroticism for females did not differ by allele group. In the total sample, Agreeableness was not associated with allele frequency; however, there was a significant allele group x clinical group x gender interaction (P<0.01). Conclusions: The present findings failed to replicate prior work suggesting that the short variant of the 5-HTTLPR allele is associated with higher Neuroticism and lower Agreeableness. © 2003 Lippincott Williams & Wilkins.

Authors
Brummett, BH; Siegler, IC; McQuoid, DR; Svenson, IK; Marchuk, DA; Steffens, DC
MLA Citation
Brummett, BH, Siegler, IC, McQuoid, DR, Svenson, IK, Marchuk, DA, and Steffens, DC. "Associations among the NEO personality inventory, revised and the serotonin transporter gene-linked polymorphic region in elders: Effects of depression and gender." Psychiatric Genetics 13.1 (2003): 13-18.
Source
scival
Published In
Psychiatric Genetics
Volume
13
Issue
1
Publish Date
2003
Start Page
13
End Page
18
DOI
10.1097/00041444-200303000-00002

Hereditary haemorrhagic telangiectasia: A questionnaire based study to delineate the different phenotypes caused by endoglin and ALK1 mutations

Background: Hereditary haemorrhagic telangiectasia (HHT) is an autosomal dominant vascular dysplasia characterised by mucocutaneous telangiectasis, epistaxis, gastrointestinal haemorrhage, and arteriovenous malformations in the lung and brain. Causative mutations for HHT have been identified in two genes, endoglin and ALK1, which encode proteins involved in serine-threonine kinase signalling in the endothelial cell. Methods: A number of people affected with HHT had completed a postal questionnaire as part of an international study to delineate the HHT phenotype. We identified questionnaires completed by subjects in whom we had identified a mutation in endoglin or ALK1. Further questionnaires were sent to families with known mutations. Data were only included from questionnaires returned by people known to carry disease causing mutations. Results: Questionnaires were completed by 83 subjects with known mutations. Of these, 49 had endoglin mutations (HHT1) and 34 had ALK1 mutations (HHT2). Subjects with HHT1 reported an earlier onset of epistaxis (p=0.01) and telangiectasis (p=0.0001) than those with HHT2. Pulmonary arteriovenous malformations were only reported in the endoglin mutation group in our study (p<0.001). Conclusions: Our questionnaire based study provides evidence that the HHT phenotype caused by mutations in endoglin (HHT1) is distinct from, and more severe than, HHT caused by mutations in ALK1 (HHT2). This has significant implications for diagnosis, screening, and treatment in the two different forms of HHT, as well as for understanding the pathogenesis of the disease.

Authors
Berg, J; Porteous, M; Reinhardt, D; Gallione, C; Holloway, S; Umasunthar, T; Lux, A; McKinnon, W; Marchuk, D; Guttmacher, A
MLA Citation
Berg, J, Porteous, M, Reinhardt, D, Gallione, C, Holloway, S, Umasunthar, T, Lux, A, McKinnon, W, Marchuk, D, and Guttmacher, A. "Hereditary haemorrhagic telangiectasia: A questionnaire based study to delineate the different phenotypes caused by endoglin and ALK1 mutations." Journal of Medical Genetics 40.8 (2003): 585-590.
PMID
12920067
Source
scival
Published In
Journal of Medical Genetics
Volume
40
Issue
8
Publish Date
2003
Start Page
585
End Page
590
DOI
10.1136/jmg.40.8.585

A kinesin heavy chain (KIF5A) mutation in hereditary spastic paraplegia (SPG10).

We have identified a missense mutation in the motor domain of the neuronal kinesin heavy chain gene KIF5A, in a family with hereditary spastic paraplegia. The mutation occurs in the family in which the SPG10 locus was originally identified, at an invariant asparagine residue that, when mutated in orthologous kinesin heavy chain motor proteins, prevents stimulation of the motor ATPase by microtubule-binding. Mutation of kinesin orthologues in various species leads to phenotypes resembling hereditary spastic paraplegia. The conventional kinesin motor powers intracellular movement of membranous organelles and other macromolecular cargo from the neuronal cell body to the distal tip of the axon. This finding suggests that the underlying pathology of SPG10 and possibly of other forms of hereditary spastic paraplegia may involve perturbation of neuronal anterograde (or retrograde) axoplasmic flow, leading to axonal degeneration, especially in the longest axons of the central nervous system.

Authors
Reid, E; Kloos, M; Ashley-Koch, A; Hughes, L; Bevan, S; Svenson, IK; Graham, FL; Gaskell, PC; Dearlove, A; Pericak-Vance, MA; Rubinsztein, DC; Marchuk, DA
MLA Citation
Reid, E, Kloos, M, Ashley-Koch, A, Hughes, L, Bevan, S, Svenson, IK, Graham, FL, Gaskell, PC, Dearlove, A, Pericak-Vance, MA, Rubinsztein, DC, and Marchuk, DA. "A kinesin heavy chain (KIF5A) mutation in hereditary spastic paraplegia (SPG10)." Am J Hum Genet 71.5 (November 2002): 1189-1194.
PMID
12355402
Source
pubmed
Published In
The American Journal of Human Genetics
Volume
71
Issue
5
Publish Date
2002
Start Page
1189
End Page
1194
DOI
10.1086/344210

Genetic modifier loci affecting survival and cardiac function in murine dilated cardiomyopathy.

BACKGROUND: Understanding the role for genetic factors in human heart failure is difficult because environmental factors cannot be standardized and genetic variation is great. One approach to identify genes that modify disease outcome is to use mouse models that show strong genetic variation of the disease phenotype. METHODS AND RESULTS: In this study, we used transgenic mice that develop severe dilated cardiomyopathy due to the cardiac-specific overexpression of calsequestrin. Transgenic mice showed marked strain-specific variation of cardiac function and survival, independent of transgene expression. A reciprocal backcross strategy was employed using two inbred strains showing distinct differences in survival and cardiac function. To map the genes that modified the heart failure phenotype, progeny from the 2 reciprocal backcrosses were used in a genome-wide scan for linkage. We identified two loci significantly linked to survival with a maximum likelihood ratio statistic of 36.2 (LOD score approximately 7.8) on chromosome 2 and of 26.5 (LOD score approximately 5.7) on chromosome 3. The chromosome 3 locus was also significantly linked to cardiac function with a maximum likelihood ratio statistic of 42.9 (LOD score approximately 9.3). Because only a single strong modifier locus was found in each backcross, we applied a haplotype analysis to map crossovers and successfully narrowed the critical intervals for each locus. CONCLUSION: Using a sensitized mouse model, we identified major modifier loci that affect the genetically complex disease of heart failure. This approach should allow the rapid identification of candidate genes involved in disease susceptibility in human populations and new insights into the pathogenesis of heart failure.

Authors
Suzuki, M; Carlson, KM; Marchuk, DA; Rockman, HA
MLA Citation
Suzuki, M, Carlson, KM, Marchuk, DA, and Rockman, HA. "Genetic modifier loci affecting survival and cardiac function in murine dilated cardiomyopathy." Circulation 105.15 (April 16, 2002): 1824-1829.
PMID
11956126
Source
pubmed
Published In
Circulation
Volume
105
Issue
15
Publish Date
2002
Start Page
1824
End Page
1829

Somatic mutation of vascular endothelial growth factor receptors in juvenile hemangioma.

Juvenile hemangiomas are the most common tumors of infancy, occurring in as many as 10% of all births. These benign vascular lesions enlarge rapidly during the first year of life by hyperplasia of endothelial cells and attendant pericytes and then spontaneously involute over a period of years, leaving loose fibrofatty tissue. Several hypotheses have been put forth concerning hemangiogenesis, including the possibility that the tumor is the result of somatic mutation in one or more components of critical vascular growth-regulatory pathways. To test this hypothesis, we obtained 15 proliferative-phase hemangiomas after surgical resection and dissected them to enrich for the lesional (endothelial and pericytic) components of each specimen. To determine whether hemangiomas represent a clonal expansion from a single progenitor cell, we assayed X-inactivation patterns for each lesion by using the polymorphic X-linked human androgen receptor gene. Twelve of 14 informative hemangiomas showed a significant degree of allelic loss after methylation-based and transcription-based polymerase chain reaction clonality assays, suggesting a nonrandom X-inactivation pattern and, thus, a monoclonal origin. We then sequenced genes encoding the receptors of the vascular endothelial growth factors (VEGFs) as candidates for potential somatic mutation. Mutations were found in two of the 15 hemangioma specimens: a missense mutation (P1147S) in the kinase domain of the VEGFR2 (FLK1/KDR) gene in one specimen and a missense mutation (P954S) in the kinase insert of the VEGFR3 (FLT4) gene in another specimen. In each case, the mutation was detected in tumor tissue but not in adjacent normal tissue. These results suggest that one potential mechanism involved in hemangioma formation is the alteration of the VEGF signaling pathway in endothelial and/or pericytic cells.

Authors
Walter, JW; North, PE; Waner, M; Mizeracki, A; Blei, F; Walker, JWT; Reinisch, JF; Marchuk, DA
MLA Citation
Walter, JW, North, PE, Waner, M, Mizeracki, A, Blei, F, Walker, JWT, Reinisch, JF, and Marchuk, DA. "Somatic mutation of vascular endothelial growth factor receptors in juvenile hemangioma." Genes Chromosomes Cancer 33.3 (March 2002): 295-303.
PMID
11807987
Source
pubmed
Published In
Genes, Chromosomes and Cancer
Volume
33
Issue
3
Publish Date
2002
Start Page
295
End Page
303

Allelic differences in the serotonin transporter-linked polymorphic region in geriatric depression.

Previous studies have examined the role of genetic variations in the serotonin transporter-linked polymorphic region (5HTTLPR) in affective disorders. The authors studied 182 older depressed subjects and 107 elderly control subjects and obtained DNA for genotyping at the 5HTTLPR. There were no significant differences in allele frequencies generally or for number of short alleles for the group as a whole, but interesting gender effects emerged. Among men, 23% of depressed men had two short alleles, compared with only 5% of control subjects. Among women, 67% of depressed women with more than one episode had at least one short allele, compared with 41% of single-episode female patients. Also, 74% of women with a positive family history of psychiatric illness in any female relative had at least one short allele, whereas 53% had at least one short allele who did not have such a family history. Our results add to the literature linking this gene to affective illness. The negative association of allele frequency and depression may be related to the relatively small sample size. The findings raise the possibility that this genetic locus may exert differential effects based on gender, increasing risk in men, and increasing risk of recurrence in women.

Authors
Steffens, DC; Svenson, I; Marchuk, DA; Levy, RM; Hays, JC; Flint, EP; Krishnan, KRR; Siegler, IC
MLA Citation
Steffens, DC, Svenson, I, Marchuk, DA, Levy, RM, Hays, JC, Flint, EP, Krishnan, KRR, and Siegler, IC. "Allelic differences in the serotonin transporter-linked polymorphic region in geriatric depression." Am J Geriatr Psychiatry 10.2 (March 2002): 185-191.
PMID
11925279
Source
pubmed
Published In
American Journal of Geriatric Psychiatry
Volume
10
Issue
2
Publish Date
2002
Start Page
185
End Page
191

KRIT1 association with the integrin-binding protein ICAP-1: a new direction in the elucidation of cerebral cavernous malformations (CCM1) pathogenesis.

Mutations in KRIT1, a protein initially identified based on a yeast two-hybrid interaction with the RAS-family GTPase RAP1A, are responsible for the development of the inherited vascular disorder cerebral cavernous malformations (CCM1). As the function of the KRIT1 protein and its role in CCM pathogenesis remain unknown, we performed yeast two-hybrid screens to identify additional protein binding partners. A fragment containing the N-terminal 272 amino acid residues of KRIT1, a region lacking similarity to any known protein upon database searches, was used as bait. From parallel screens of human fetal brain and HeLa cDNA libraries, we obtained multiple independent isolates of human integrin cytoplasmic domain-associated protein-1 (ICAP-1) as interacting clones. The interaction of KRIT1 and ICAP-1 was confirmed by GST-KRIT1 trapping of endogenous ICAP-1 from 293T cells. The alpha isoform of ICAP-1 is a 200 amino acid serine/threonine-rich phosphoprotein which binds the cytoplasmic tail of beta1 integrins. We show that mutagenesis of the N-terminal KRIT1 NPXY amino acid sequence, a motif critical for ICAP-1 binding to beta1 integrin molecules, completely abrogates the KRIT1/ICAP-1 interaction. The interaction between ICAP-1 and KRIT1, and the presence of a FERM domain in the latter, suggest that KRIT1 might be involved in the bidirectional signaling between integrin molecules and the cytoskeleton. Furthermore, these data suggest that KRIT1 might affect cell adhesion processes via integrin signaling in CCM1 pathogenesis.

Authors
Zawistowski, JS; Serebriiskii, IG; Lee, MF; Golemis, EA; Marchuk, DA
MLA Citation
Zawistowski, JS, Serebriiskii, IG, Lee, MF, Golemis, EA, and Marchuk, DA. "KRIT1 association with the integrin-binding protein ICAP-1: a new direction in the elucidation of cerebral cavernous malformations (CCM1) pathogenesis." Hum Mol Genet 11.4 (February 15, 2002): 389-396.
PMID
11854171
Source
pubmed
Published In
Human Molecular Genetics
Volume
11
Issue
4
Publish Date
2002
Start Page
389
End Page
396

A second leaky splice-site mutation in the spastin gene.

Authors
Svenson, IK; Ashley-Koch, AE; Pericak-Vance, MA; Marchuk, DA
MLA Citation
Svenson, IK, Ashley-Koch, AE, Pericak-Vance, MA, and Marchuk, DA. "A second leaky splice-site mutation in the spastin gene." Am J Hum Genet 69.6 (December 2001): 1407-1409. (Letter)
PMID
11704932
Source
pubmed
Published In
The American Journal of Human Genetics
Volume
69
Issue
6
Publish Date
2001
Start Page
1407
End Page
1409
DOI
10.1086/324593

Identification and expression analysis of spastin gene mutations in hereditary spastic paraplegia.

Pure hereditary spastic paraplegia (SPG) type 4 is the most common form of autosomal dominant hereditary SPG, a neurodegenerative disease characterized primarily by hyperreflexia and progressive spasticity of the lower limbs. It is caused by mutations in the gene encoding spastin, a member of the AAA family of ATPases. We have screened the spastin gene for mutations in 15 families consistent with linkage to the spastin gene locus, SPG4, and have identified 11 mutations, 10 of which are novel. Five of the mutations identified are in noninvariant splice-junction sequences. Reverse transcription-PCR analysis of mRNA from patients shows that each of these five mutations results in aberrant splicing. One mutation was found to be "leaky," or partially penetrant; that is, the mutant allele produced both mutant (skipped exon) and wild-type (full-length) transcripts. This phenomenon was reproduced in in vitro splicing experiments, with a minigene splicing-vector construct only in the context of the endogenous splice junctions flanking the splice junctions of the skipped exon. In the absence of endogenous splice junctions, only mutant transcript was detected. The existence of at least one leaky mutation suggests that relatively small differences in the level of wild-type spastin expression can have significant functional consequences. This may account, at least in part, for the wide ranges in age at onset, symptom severity, and rate of symptom progression that have been reported to occur both among and within families with SPG linked to SPG4. In addition, these results suggest caution in the interpretation of data solely obtained with minigene constructs to study the effects of sequence variation on splicing. The lack of full genomic sequence context in these constructs can mask important functional consequences of the mutation.

Authors
Svenson, IK; Ashley-Koch, AE; Gaskell, PC; Riney, TJ; Cumming, WJ; Kingston, HM; Hogan, EL; Boustany, RM; Vance, JM; Nance, MA; Pericak-Vance, MA; Marchuk, DA
MLA Citation
Svenson, IK, Ashley-Koch, AE, Gaskell, PC, Riney, TJ, Cumming, WJ, Kingston, HM, Hogan, EL, Boustany, RM, Vance, JM, Nance, MA, Pericak-Vance, MA, and Marchuk, DA. "Identification and expression analysis of spastin gene mutations in hereditary spastic paraplegia." Am J Hum Genet 68.5 (May 2001): 1077-1085.
PMID
11309678
Source
pubmed
Published In
The American Journal of Human Genetics
Volume
68
Issue
5
Publish Date
2001
Start Page
1077
End Page
1085
DOI
10.1086/320111

Pathogenesis of hemangioma.

Authors
Marchuk, DA
MLA Citation
Marchuk, DA. "Pathogenesis of hemangioma." J Clin Invest 107.6 (March 2001): 665-666. (Review)
PMID
11254664
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
107
Issue
6
Publish Date
2001
Start Page
665
End Page
666
DOI
10.1172/JCI12470

Evidence for loss of heterozygosity of 5q in sporadic haemangiomas: are somatic mutations involved in haemangioma formation?

BACKGROUND/AIMS: Haemangiomas are common benign tumours of infancy that consist of rapidly proliferating endothelial cells. A locus for an autosomal dominant predisposition to haemangioma has been identified recently on chromosome 5q. This study aimed to investigate loss of heterozygosity on chromosomes 5 and 9 in haemangiomas. METHODS: Sporadic proliferative phase haemangiomas were microdissected. Polymerase chain reaction amplification and analysis of microsatellite markers on chromosomes 5 and 9 was carried out. RESULTS: There was a significant loss of heterozygosity for markers on chromosome 5q in haemangioma tissue, when compared with either markers from chromosome 5p (p < 0.05) or markers from chromosome 9 (p < 0.05). CONCLUSIONS: These results suggest that haemangioma formation might be associated with somatic mutational events, and provides evidence that a locus on 5q is involved in the formation of sporadic haemangiomas.

Authors
Berg, JN; Walter, JW; Thisanagayam, U; Evans, M; Blei, F; Waner, M; Diamond, AG; Marchuk, DA; Porteous, ME
MLA Citation
Berg, JN, Walter, JW, Thisanagayam, U, Evans, M, Blei, F, Waner, M, Diamond, AG, Marchuk, DA, and Porteous, ME. "Evidence for loss of heterozygosity of 5q in sporadic haemangiomas: are somatic mutations involved in haemangioma formation?." J Clin Pathol 54.3 (March 2001): 249-252.
PMID
11253142
Source
pubmed
Published In
Journal of Clinical Pathology
Volume
54
Issue
3
Publish Date
2001
Start Page
249
End Page
252

Fine mapping and genetic heterogeneity in the pure form of autosomal dominant familial spastic paraplegia.

We evaluated seven families segregating pure, autosomal dominant familial spastic paraplegia (SPG) for linkage to four recently identified SPG loci on chromosomes 2q (1), 8q (2), 12q (3), and 19q (4). These families were previously shown to be unlinked to SPG loci on chromosomes 2p, 14q, and 15q. Two families demonstrated linkage to the new loci. One family (family 3) showed significant evidence for linkage to chromosome 12q, peaking at D12S1691 (maximum lod = 3.22). Haplotype analysis of family 3 did not identify any recombinants among affected individuals in the 12q candidate region. Family 5 yielded a peak lod score of 2.02 at marker D19S868 and excluded linkage to other known SPG loci. Haplotype analysis of family 5 revealed several cross-overs in affected individuals, thereby potentially narrowing the SPG12 candidate region to a 5-cM region between markers D19S868 and D19S220. Three of the families definitively excluded all four loci examined, providing evidence for further genetic heterogeneity of pure, autosomal dominant SPG. In conclusion, these data confirm the presence of SPG10 (chromosome 12), potentially reduce the minimum candidate region for SPG12 (chromosome 19q), and suggest there is at least one additional autosomal dominant SPG locus.

Authors
Ashley-Koch, A; Bonner, ER; Gaskell, PC; West, SG; Tim, R; Wolpert, CM; Jones, R; Farrell, CD; Nance, M; Svenson, IK; Marchuk, DA; Boustany, RM; Vance, JM; Scott, WK; Pericak-Vance, MA
MLA Citation
Ashley-Koch, A, Bonner, ER, Gaskell, PC, West, SG, Tim, R, Wolpert, CM, Jones, R, Farrell, CD, Nance, M, Svenson, IK, Marchuk, DA, Boustany, RM, Vance, JM, Scott, WK, and Pericak-Vance, MA. "Fine mapping and genetic heterogeneity in the pure form of autosomal dominant familial spastic paraplegia." Neurogenetics 3.2 (March 2001): 91-97.
PMID
11354831
Source
pubmed
Published In
Neurogenetics
Volume
3
Issue
2
Publish Date
2001
Start Page
91
End Page
97

Central nervous system serotonin function and cardiovascular responses to stress.

OBJECTIVE: The objective of this study was to evaluate the impact of indices of central nervous system (CNS) serotonin function on cardiovascular reactivity to mental stress. METHODS: Lumbar puncture was performed on 54 healthy volunteers to obtain cerebrospinal fluid (CSF) for determination of 5-hydroxyindoleacetic acid (5HIAA) levels. Genotypes were determined with respect to a functional polymorphism of the serotonin transporter gene promoter region (5HTTLPR). Subjects then underwent mental stress testing. RESULTS: Persons with one or two long (l) 5HTTLPR alleles had CSF levels of the major serotonin metabolite, 5HIAA, that were 50% higher than those of persons with the s/s 5HTTLPR genotype. Persons with one or two l alleles or higher CSF 5HIAA levels also exhibited greater blood pressure and heart rate responses to a mental stress protocol. CONCLUSIONS: These findings suggest the 5HTTLPR polymorphism affects CNS serotonin function, and they are consistent with the general hypothesis that CNS serotonin function is involved in the regulation of potentially health-damaging biobehavioral characteristics. In particular, the l allele could contribute, through its association with increased cardiovascular reactivity to stress, to increased risk of cardiovascular disease.

Authors
Williams, RB; Marchuk, DA; Gadde, KM; Barefoot, JC; Grichnik, K; Helms, MJ; Kuhn, CM; Lewis, JG; Schanberg, SM; Stafford-Smith, M; Suarez, EC; Clary, GL; Svenson, IK; Siegler, IC
MLA Citation
Williams, RB, Marchuk, DA, Gadde, KM, Barefoot, JC, Grichnik, K, Helms, MJ, Kuhn, CM, Lewis, JG, Schanberg, SM, Stafford-Smith, M, Suarez, EC, Clary, GL, Svenson, IK, and Siegler, IC. "Central nervous system serotonin function and cardiovascular responses to stress." Psychosom Med 63.2 (March 2001): 300-305.
PMID
11292279
Source
pubmed
Published In
Psychosomatic Medicine
Volume
63
Issue
2
Publish Date
2001
Start Page
300
End Page
305

Computational and experimental analyses reveal previously undetected coding exons of the KRIT1 (CCM1) gene.

A notable difficulty in annotating genomic sequence is identifying the correct start codon in a gene. An important such case has been found with KRIT1, the cerebral cavernous malformation type 1 (CCM1) gene. Analysis of human and mouse genomic sequence encompassing the region containing KRIT1/Krit1 using exon/gene-prediction and comparative alignment programs revealed putative exons upstream of the previously described first exon. These additional candidate exons show significant matches to mouse and human ESTs that are contiguous with and extend upstream from the previously designated 5' end of the KRIT1 cDNA sequence. RT-PCR and 5'RACE experiments confirm the presence of four additional upstream coding exons that encode an additional 207 amino acids. Importantly, a novel frameshift mutation in one of these newly identified KRIT1 exons has been found in a CCM1 family. These data establish the authentic KRIT1 amino acid sequence and suggest that the additional KRIT1 exons may harbor mutations in other CCM1 families. In addition, these results provide another example of the utility of rigorous computational and comparative sequence analysis for refining gene structure.

Authors
Sahoo, T; Goenaga-Diaz, E; Serebriiskii, IG; Thomas, JW; Kotova, E; Cuellar, JG; Peloquin, JM; Golemis, E; Beitinjaneh, F; Green, ED; Johnson, EW; Marchuk, DA
MLA Citation
Sahoo, T, Goenaga-Diaz, E, Serebriiskii, IG, Thomas, JW, Kotova, E, Cuellar, JG, Peloquin, JM, Golemis, E, Beitinjaneh, F, Green, ED, Johnson, EW, and Marchuk, DA. "Computational and experimental analyses reveal previously undetected coding exons of the KRIT1 (CCM1) gene." Genomics 71.1 (January 1, 2001): 123-126.
PMID
11161805
Source
pubmed
Published In
Genomics
Volume
71
Issue
1
Publish Date
2001
Start Page
123
End Page
126
DOI
10.1006/geno.2000.6426

Additional glomangioma families link to chromosome 1p: no evidence for genetic heterogeneity.

Venous malformations are a common abnormality of the vasculature that may occur sporadically or, more rarely, as an autosomal dominant trait. One familial form of venous malformations has previously been linked to chromosome 9p. Mutations in the gene encoding Tie2, an endothelial specific receptor tyrosine kinase, have been identified in four different families. Glomangiomas are a subtype of venous malformations with glomus cell involvement. These cutaneous lesions can be inherited as an autosomal dominant disease with reduced penetrance and variable expressivity. We present evidence of linkage to chromosome 1p21-1p22 using four new glomangioma families, with a combined maximum two-point lod score of 7.32 at marker D1S2804. Markers D1S2129 and D1S2881 define the 24-cM linkage interval determined by recombination within affected individuals. A recent report also showed linkage of the glomangioma locus to chromosome 1p. A total of 9 families now map to this region, suggesting a decreased likelihood of locus heterogenity in familial glomangiomas. Investigation of candidate genes within the interval should provide new insights into lesion formation in inherited venous malformations.

Authors
Calvert, JT; Burns, S; Riney, TJ; Sahoo, T; Orlow, SJ; Nevin, NC; Haisley-Royster, C; Prose, N; Simpson, SA; Speer, MC; Marchuk, DA
MLA Citation
Calvert, JT, Burns, S, Riney, TJ, Sahoo, T, Orlow, SJ, Nevin, NC, Haisley-Royster, C, Prose, N, Simpson, SA, Speer, MC, and Marchuk, DA. "Additional glomangioma families link to chromosome 1p: no evidence for genetic heterogeneity." Hum Hered 51.3 (2001): 180-182.
PMID
11173970
Source
pubmed
Published In
Human heredity
Volume
51
Issue
3
Publish Date
2001
Start Page
180
End Page
182

Abnormal pattern of Tie-2 and vascular endothelial growth factor receptor expression in human cerebral arteriovenous malformations.

OBJECTIVE: Human cerebral arteriovenous malformations (AVMs) are speculated to result from abnormal angiogenesis. Vascular endothelial growth factor receptors (VEGF-Rs) and Tie-2 play critical roles in vasculogenesis and angiogenesis. We hypothesized that the abnormal vascular phenotype of AVMs may be associated with abnormal expression of VEGF-Rs and Tie-2. METHODS: We measured the expression of Tie-2, VEGF-R1, and VEGF-R2 in AVMs and normal brain tissue, using immunoblotting. To assess active vascular remodeling, we also measured endothelial nitric oxide synthase expression. CD31 expression was used to control for endothelial cell mass for Tie-2, VEGF-Rs, and endothelial nitric oxide synthase. Immunoblotting data were presented as relative expression, using normal brain tissue values as 100%. RESULTS: CD31 was expressed to similar degrees in AVMs and normal brain tissue (99+/-29% versus 100+/-20%, mean +/- standard error, P = 0.98). Tie-2 expression was markedly decreased in all AVMs, compared with normal brain tissue (16+/-9% versus 100+/-37%, P = 0.04). VEGF-R1 expression was decreased in four of five AVMs, but the difference between the mean values was not significant (35+/-8% versus 100+/-42%, P = 0.14). VEGF-R2 expression was decreased in all AVMs, compared with normal brain tissue (28+/-6% versus 100+/-29%, P = 0.03). There was no difference in endothelial nitric oxide synthase expression between AVMs and normal brain tissue (106+/-42% versus 100+/-25%, P = 0.91). CONCLUSION: AVM vessels exhibited abnormal expression of Tie-2 and VEGF-Rs, both of which may contribute to the pathogenesis of AVMs.

Authors
Hashimoto, T; Emala, CW; Joshi, S; Mesa-Tejada, R; Quick, CM; Feng, L; Libow, A; Marchuk, DA; Young, WL
MLA Citation
Hashimoto, T, Emala, CW, Joshi, S, Mesa-Tejada, R, Quick, CM, Feng, L, Libow, A, Marchuk, DA, and Young, WL. "Abnormal pattern of Tie-2 and vascular endothelial growth factor receptor expression in human cerebral arteriovenous malformations." Neurosurgery 47.4 (October 2000): 910-918.
PMID
11014431
Source
pubmed
Published In
Neurosurgery
Volume
47
Issue
4
Publish Date
2000
Start Page
910
End Page
918

Clinical manifestations in a large hereditary hemorrhagic telangiectasia (HHT) type 2 kindred.

HHT type 2 (HHT 2) is a multi-system vascular dysplasia caused by a mutation in the ALK-1 gene, but the phenotype has not been well defined. We report on 51 members of an HHT 2 kindred with an ALK-1 gene mutation shown to be associated with the disorder. This ALK-1 mutation was detected in 38 kindred members who were evaluated systematically for associated vascular abnormalities. Pulmonary arteriovenous malformations (AVMs) were found in 6% of those screened, cerebral AVM in 7%, hepatic AVM in 17%, and spinal AVM in 3%. We discuss these and other findings in the 38 affected kindred members, as well as findings in the 13 kindred members in whom the mutation was not detected. This study shows that pulmonary, cerebral, spinal, and hepatic AVMs can all occur in HHT 2. It also adds to the evidence suggesting that pulmonary AVMs are more common in HHT 1 than in HHT 2. We identify a higher prevalence of hepatic AVMs than previously reported in either HHT 1 or 2. This may be specific to the mutation in this kindred, but probably reflects the lack of routine screening for this manifestation. Even in this family in which all affected individuals have the same mutation, the clinical manifestations of HHT and their severity varied tremendously. Intrafamilial variation in expression of HHT is clearly significant, emphasizing the difficulty in establishing the diagnosis in individuals and in sub-typing families when DNA testing is not available.

Authors
McDonald, JE; Miller, FJ; Hallam, SE; Nelson, L; Marchuk, DA; Ward, KJ
MLA Citation
McDonald, JE, Miller, FJ, Hallam, SE, Nelson, L, Marchuk, DA, and Ward, KJ. "Clinical manifestations in a large hereditary hemorrhagic telangiectasia (HHT) type 2 kindred." Am J Med Genet 93.4 (August 14, 2000): 320-327. (Review)
PMID
10946360
Source
pubmed
Published In
American Journal of Medical Genetics Part A
Volume
93
Issue
4
Publish Date
2000
Start Page
320
End Page
327

Two common endoglin mutations in families with hereditary hemorrhagic telangiectasia in the Netherlands Antilles: evidence for a founder effect.

Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant bleeding disorder characterized by localized angiodysplasia. Mutations in either of two genes, endoglin or ALK-1, can cause HHT. Both genes encode putative receptors for the transforming growth factor-beta superfamily of ligands. Many mutations in each gene have been identified in HHT kindreds from around the world, and with few exceptions mutations are unique and family specific. The prevalence of HHT in the Leeward Islands of the Netherlands Antilles is possibly the highest of any geographical location. We wished to establish whether this high prevalence is due to a genetic founder effect or to multiple mutational events. HHT kindreds from the Netherlands Antilles and The Netherlands were screened for mutations in the two genes associated with HHT. Haplotype analysis of a 5-cM region on chromosome 9 flanking the endoglin gene revealed three distinct disease haplotypes in the ten Antillean families studied. Seven of these families share a splice-site mutation in exon 1 of endoglin. Two other Antillean families share a missense mutation in exon 9a of endoglin. This mutation was also found in a Dutch family that shares the same disease haplotype as the Antillean families with this mutation. Thus it appears that HHT in the Netherlands Antilles is due to a limited number of ancestral mutations in the endoglin gene, and that one of these mutations was introduced into the African slave population by a Dutch colonist. The limited scope of mutations suggests that a presymptomatic screening program for HHT would be feasible in this population.

Authors
Gallione, CJ; Scheessele, EA; Reinhardt, D; Duits, AJ; Berg, JN; Westermann, CJ; Marchuk, DA
MLA Citation
Gallione, CJ, Scheessele, EA, Reinhardt, D, Duits, AJ, Berg, JN, Westermann, CJ, and Marchuk, DA. "Two common endoglin mutations in families with hereditary hemorrhagic telangiectasia in the Netherlands Antilles: evidence for a founder effect." Hum Genet 107.1 (July 2000): 40-44.
PMID
10982033
Source
pubmed
Published In
Human Genetics
Volume
107
Issue
1
Publish Date
2000
Start Page
40
End Page
44

Expression analysis of endoglin missense and truncation mutations: insights into protein structure and disease mechanisms.

Hereditary hemorrhagic telangiectasia (HHT) is an inherited autosomal dominant vascular dysplasia caused by mutations in either endoglin (HHT1) or activin-like kinase receptor-1 (ALK-1) (HHT2). The majority of the mutations in endoglin cause frameshifts and premature stop codons. Although initial reports suggested a dominant-negative model for HHT1, more recent reports have suggested that mutations in endoglin lead to haploinsufficiency. In this study, we investigated six different missense mutations and two truncation mutations in the endoglin gene to examine whether mechanisms other than haploinsufficiency might be involved in HHT1. Expression of the missense mutants alone revealed that they are misfolded and that most show no cell surface expression. When co-expressed with wild-type endoglin, the missense mutants are able to dimerize with the normal endoglin protein and are trafficked to the cell surface. We also show that although one truncation mutation acts through haploinsufficiency, the other acts in a dominant-negative way. This implies that either dominant-negative protein interactions or haploinsufficiency can cause HHT1. The biochemical analyses for the different mutations suggest that the endoglin N-terminus is important for correct protein folding and that cysteine residues in the first 350 amino acids are involved in intramolecular disulfide bonds, whereas cysteines located closer to the C-terminus of the extracellular domain are responsible for inter-molecular disulfide bond dimerization.

Authors
Lux, A; Gallione, CJ; Marchuk, DA
MLA Citation
Lux, A, Gallione, CJ, and Marchuk, DA. "Expression analysis of endoglin missense and truncation mutations: insights into protein structure and disease mechanisms." Hum Mol Genet 9.5 (March 22, 2000): 745-755.
PMID
10749981
Source
pubmed
Published In
Human Molecular Genetics
Volume
9
Issue
5
Publish Date
2000
Start Page
745
End Page
755

Endoglin, an ancillary TGFbeta receptor, is required for extraembryonic angiogenesis and plays a key role in heart development.

Endoglin (CD105) is expressed on the surface of endothelial and haematopoietic cells in mammals and binds TGFbeta isoforms 1 and 3 in combination with the signaling complex of TGFbeta receptors types I and II. Endoglin expression increases during angiogenesis, wound healing, and inflammation, all of which are associated with TGFbeta signaling and alterations in vascular structure. The importance of endoglin for normal vascular architecture is further indicated by the association of mutations in the endoglin gene with the inherited disorder Hereditary Haemorrhagic Telangiectasia Type 1 (HHT1), a disease characterised by bleeding from vascular malformations. In order to study the role of endoglin in vivo in more detail and to work toward developing an animal model of HHT1, we have derived mice that carry a targeted nonsense mutation in the endoglin gene. Studies on these mice have revealed that endoglin is essential for early development. Embryos homozygous for the endoglin mutation fail to progress beyond 10.5 days postcoitum and fail to form mature blood vessels in the yolk sac. This phenotype is remarkably similar to that of the TGFbeta1 and the TGFbeta receptor II knockout mice, indicating that endoglin is needed in vivo for TGFbeta1 signaling during extraembryonic vascular development. In addition, we have observed cardiac defects in homozygous endoglin-deficient embryos, suggesting endoglin also plays a role in cardiogenesis. We anticipate that heterozygous mice will ultimately serve as a useful disease model for HHT1, as some individuals have dilated and fragile blood vessels similar to vascular malformations seen in HHT patients.

Authors
Arthur, HM; Ure, J; Smith, AJ; Renforth, G; Wilson, DI; Torsney, E; Charlton, R; Parums, DV; Jowett, T; Marchuk, DA; Burn, J; Diamond, AG
MLA Citation
Arthur, HM, Ure, J, Smith, AJ, Renforth, G, Wilson, DI, Torsney, E, Charlton, R, Parums, DV, Jowett, T, Marchuk, DA, Burn, J, and Diamond, AG. "Endoglin, an ancillary TGFbeta receptor, is required for extraembryonic angiogenesis and plays a key role in heart development." Dev Biol 217.1 (January 1, 2000): 42-53.
PMID
10625534
Source
pubmed
Published In
Developmental Biology
Volume
217
Issue
1
Publish Date
2000
Start Page
42
End Page
53
DOI
10.1006/dbio.1999.9534

Expression analysis of four endoglin missense mutations suggests that haploinsufficiency is the predominant mechanism for hereditary hemorrhagic telangiectasia type 1.

ENDOGLIN codes for a homodimeric membrane glycoprotein that interacts with receptors for members of the TGF-beta superfamily and is the gene mutated in the autosomal dominant vascular disorder hereditary hemorrhagic telangiectasia type 1 (HHT1). We recently demonstrated that functional endoglin was expressed at half levels on human umbilical vein endothelial cells (HUVECs) and peripheral blood activated monocytes from HHT1 patients. Two types of mutant protein were previously analyzed, the product of an exon 3 skip which was expressed as a transient intracellular species and prematurely truncated proteins that were undetectable in patient samples. Here we report the analysis of four proteins resulting from point mutations, with missense codons G52V and C53R in exon 2, W149C in exon 4 and L221P in exon 5. Metabolic labeling of activated monocytes from confirmed, clinically affected patients revealed reduced expression of fully processed normal endoglin in all cases. Pulse-chase analysis with HUVECs from a newborn with the C53R substitution indicated that mutant endoglin remained intracellular as a precursor form and did not impair processing of the normal protein. Biotinylation of cell surface proteins, metabolic labeling and pulse-chase analysis revealed that none of the engineered missense mutants was significantly expressed at the surface of COS-1 transfectants. Thus, these four HHT1 missense mutations lead to transient intracellular species which cannot interfere with normal endoglin function. These data suggest that haploinsufficiency, leading to reduced levels of one of the major surface glyco-proteins of vascular endothelium, is the predominant mechanism underlying the HHT1 phenotype.

Authors
Pece-Barbara, N; Cymerman, U; Vera, S; Marchuk, DA; Letarte, M
MLA Citation
Pece-Barbara, N, Cymerman, U, Vera, S, Marchuk, DA, and Letarte, M. "Expression analysis of four endoglin missense mutations suggests that haploinsufficiency is the predominant mechanism for hereditary hemorrhagic telangiectasia type 1." Hum Mol Genet 8.12 (November 1999): 2171-2181.
PMID
10545596
Source
pubmed
Published In
Human Molecular Genetics
Volume
8
Issue
12
Publish Date
1999
Start Page
2171
End Page
2181

Mutations in the gene encoding KRIT1, a Krev-1/rap1a binding protein, cause cerebral cavernous malformations (CCM1).

Cerebral cavernous malformations (CCM) are congenital vascular anomalies of the brain that can cause significant neurological disabilities, including intractable seizures and hemorrhagic stroke. One locus for autosomal dominant CCM ( CCM1 ) maps to chromosome 7q21-q22. Recombination events in linked family members define a critical region of approximately 2 Mb and a shared disease haplotype associated with a presumed founder effect in families of Mexican-American descent points to a potentially smaller region of interest. Using a genomic sequence-based positional cloning strategy, we have identified KRIT1, encoding a protein that interacts with the Krev-1/rap1a tumor suppressor, as the CCM1 gene. Seven different KRIT1 mutations have been identified in 23 distinct CCM1 families. The identical mutation is present in 16 of 21 Mexican-American families analyzed, substantiating a founder effect in this population. Other Mexican-American and non-Hispanic Caucasian CCM1 kindreds harbor other KRIT1 mutations. Identification of a common Mexican-American mutation has potential clinical significance for presymptomatic diagnosis of CCM in this population. In addition, these data point to a key role for the Krev-1/rap1a signaling pathway in angiogenesis and cerebrovascular disease.

Authors
Sahoo, T; Johnson, EW; Thomas, JW; Kuehl, PM; Jones, TL; Dokken, CG; Touchman, JW; Gallione, CJ; Lee-Lin, SQ; Kosofsky, B; Kurth, JH; Louis, DN; Mettler, G; Morrison, L; Gil-Nagel, A; Rich, SS; Zabramski, JM; Boguski, MS; Green, ED; Marchuk, DA
MLA Citation
Sahoo, T, Johnson, EW, Thomas, JW, Kuehl, PM, Jones, TL, Dokken, CG, Touchman, JW, Gallione, CJ, Lee-Lin, SQ, Kosofsky, B, Kurth, JH, Louis, DN, Mettler, G, Morrison, L, Gil-Nagel, A, Rich, SS, Zabramski, JM, Boguski, MS, Green, ED, and Marchuk, DA. "Mutations in the gene encoding KRIT1, a Krev-1/rap1a binding protein, cause cerebral cavernous malformations (CCM1)." Hum Mol Genet 8.12 (November 1999): 2325-2333.
PMID
10545614
Source
pubmed
Published In
Human Molecular Genetics
Volume
8
Issue
12
Publish Date
1999
Start Page
2325
End Page
2333

Allelic and locus heterogeneity in inherited venous malformations.

Venous malformations are low-flow vascular lesions consisting of disorganized thin-walled vascular channels. These can occur sporadically but also as an autosomal dominant condition termed venous malformations, cutaneous and mucosal (VMCM; OMIM 600195). In two large unrelated kindreds mapping to chromosome 9, the identical R849W missense mutation was identified in the first kinase domain of Tie2, an endothelial cell-specific receptor tyrosine kinase. We report here the identification of four new kindreds with inherited venous malformations. Unlike the initial two families described, these four families demonstrate allelic and locus heterogeneity. In one of these families, the R849W mutation co-segregates with the disease phenotype. Three other families with venous malformations lack this mutation. One of these families is linked to markers near TIE2 on chromosome 9. In this family, we identified a novel mutation within the first kinase domain of Tie2 resulting in a Y897S change. Results from COS-1 cell transfections using expression constructs containing either the R849W or the Y897S mutation suggest that the receptors containing either mutation show ligand-independent hyperphosphorylation. These results suggest a gain-of-function mechanism for development of venous malformations in these families. Of the two remaining families, one excludes linkage to the TIE2 locus, establishing the existence of at least one additional locus for dominantly inherited venous malformations.

Authors
Calvert, JT; Riney, TJ; Kontos, CD; Cha, EH; Prieto, VG; Shea, CR; Berg, JN; Nevin, NC; Simpson, SA; Pasyk, KA; Speer, MC; Peters, KG; Marchuk, DA
MLA Citation
Calvert, JT, Riney, TJ, Kontos, CD, Cha, EH, Prieto, VG, Shea, CR, Berg, JN, Nevin, NC, Simpson, SA, Pasyk, KA, Speer, MC, Peters, KG, and Marchuk, DA. "Allelic and locus heterogeneity in inherited venous malformations." Hum Mol Genet 8.7 (July 1999): 1279-1289.
PMID
10369874
Source
pubmed
Published In
Human Molecular Genetics
Volume
8
Issue
7
Publish Date
1999
Start Page
1279
End Page
1289

Assignment of transforming growth factor beta1 and beta3 and a third new ligand to the type I receptor ALK-1.

Germ line mutations in one of two distinct genes, endoglin or ALK-1, cause hereditary hemorrhagic telangiectasia (HHT), an autosomal dominant disorder of localized angiodysplasia. Both genes encode endothelial cell receptors for the transforming growth factor beta (TGF-beta) ligand superfamily. Endoglin has homology to the type III receptor, betaglycan, although its exact role in TGF-beta signaling is unclear. Activin receptor-like kinase 1 (ALK-1) has homology to the type I receptor family, but its ligand and corresponding type II receptor are unknown. In order to identify the ligand and type II receptor for ALK-1 and to investigate the role of endoglin in ALK-1 signaling, we devised a chimeric receptor signaling assay by exchanging the kinase domain of ALK-1 with either the TGF-beta type I receptor or the activin type IB receptor, both of which can activate an inducible PAI-1 promoter. We show that TGF-beta1 and TGF-beta3, as well as a third unknown ligand present in serum, can activate chimeric ALK-1. HHT-associated missense mutations in the ALK-1 extracellular domain abrogate signaling. The ALK-1/ligand interaction is mediated by the type II TGF-beta receptor for TGF-beta and most likely through the activin type II or type IIB receptors for the serum ligand. Endoglin is a bifunctional receptor partner since it can bind to ALK-1 as well as to type I TGF-beta receptor. These data suggest that HHT pathogenesis involves disruption of a complex network of positive and negative angiogenic factors, involving TGF-beta, a new unknown ligand, and their corresponding receptors.

Authors
Lux, A; Attisano, L; Marchuk, DA
MLA Citation
Lux, A, Attisano, L, and Marchuk, DA. "Assignment of transforming growth factor beta1 and beta3 and a third new ligand to the type I receptor ALK-1." J Biol Chem 274.15 (April 9, 1999): 9984-9992.
PMID
10187774
Source
pubmed
Published In
The Journal of biological chemistry
Volume
274
Issue
15
Publish Date
1999
Start Page
9984
End Page
9992

Genetic mapping of a novel familial form of infantile hemangioma.

Infantile hemangiomas are the most common tumor of infancy, occurring with an incidence of up to 10% of all births. They are benign but highly proliferative lesions involving aberrant localized growth of capillary endothelium. Although most hemangiomas occur sporadically and as single lesions, or in conjunction with pleiotropic genetic syndromes, we have previously identified six kindreds where hemangiomas appear to segregate as an autosomal dominant trait with high penetrance. Four such families contain affected individuals in three or more generations. In the current study, blood samples from five of these families were collected and used in a whole genome linkage search at 10-cM resolution. We established evidence for linkage to 5q in three families, and evidence for locus heterogeneity. The three 5q-linked families were further genotyped to generate haplotype information and narrow the candidate interval. Based on recombination breakpoint analysis, the interval exists between markers D5S2490 and D5S408, spanning 55 cM, and corresponding to 5q31-33. Using information from affected and unaffected individuals, the interval spans 38 cM between markers D5S1469 and D5S211. Three candidate genes involved with blood vessel growth map to this region: fibroblast growth factor receptor-4 (FGFR4), platelet-derived growth factor receptor-beta (PDG-FRB), and fms-related tyrosine kinase-4 (FLT4). The genes and gene products associated with familial hemangiomas may be involved somatically in the more common sporadic cases.

Authors
Walter, JW; Blei, F; Anderson, JL; Orlow, SJ; Speer, MC; Marchuk, DA
MLA Citation
Walter, JW, Blei, F, Anderson, JL, Orlow, SJ, Speer, MC, and Marchuk, DA. "Genetic mapping of a novel familial form of infantile hemangioma." Am J Med Genet 82.1 (January 1, 1999): 77-83.
PMID
9916848
Source
pubmed
Published In
American Journal of Medical Genetics Part A
Volume
82
Issue
1
Publish Date
1999
Start Page
77
End Page
83

Cloning of the promoter region of human endoglin, the target gene for hereditary hemorrhagic telangiectasia type 1.

Endoglin (CD105) is a cell surface component of the transforming growth factor-beta (TGF-beta) receptor complex highly expressed by endothelial cells. Mutations in the endoglin gene are responsible for the hereditary hemorrhagic telangiectasia type 1 (HHT1), also known as Osler-Weber-Rendu syndrome (OMIM 187300). This is an autosomal dominant vascular disorder probably caused by a haploinsufficiency mechanism displaying low levels of the normal protein. To understand the mechanisms underlying the regulated expression of endoglin, a genomic DNA clone containing 3.3 kb of the 5'-flanking sequence of the human endoglin gene has been isolated. The 5'-flanking region of the endoglin gene lacks consensus TATA and CAAT boxes, but contains two GC-rich regions and consensus motifs for Sp1, ets, GATA, AP-2, NFkappaB, and Mad, as well as TGF-beta-, glucocorticoid-, vitamin D-, and estrogen-responsive elements. As determined by primer extension and 5' RACE experiments, a cluster of transcriptional start sites was found to be located 350 bp upstream from the translation initiation codon. To analyze the endoglin promoter activity, the upstream -400/+341 fragment was fused to the luciferase gene and transient transfections were conducted in several cell types. This construct displayed a tissue-specific activity in human and bovine endothelial cells. Analysis of various deletion constructs showed the existence of a basal promoter region within the -81/+350 fragment as well as major transcriptional regulatory elements within the -400/-141 fragment. Electrophoretic mobility shift assays demonstrated the specific interaction of a member of the ets family with a consensus motif located at position -68. A promoter construct mutated at this ets sequence showed a much reduced activity as compared with the wild-type construct, supporting the involvement of this ets motif in the basal activity of the promoter. The endoglin promoter exhibited inducibility in the presence of TGF-beta1, suggesting possible therapeutic treatments in HHT1 patients, in which the expression level of the normal endoglin allele might not reach the threshold required for its function. Isolation and characterization of the human endoglin promoter represents an initial step in elucidating the controlled expression of the endoglin gene.

Authors
Ríus, C; Smith, JD; Almendro, N; Langa, C; Botella, LM; Marchuk, DA; Vary, CP; Bernabéu, C
MLA Citation
Ríus, C, Smith, JD, Almendro, N, Langa, C, Botella, LM, Marchuk, DA, Vary, CP, and Bernabéu, C. "Cloning of the promoter region of human endoglin, the target gene for hereditary hemorrhagic telangiectasia type 1." Blood 92.12 (December 15, 1998): 4677-4690.
PMID
9845534
Source
pubmed
Published In
Blood
Volume
92
Issue
12
Publish Date
1998
Start Page
4677
End Page
4690

Genetic abnormalities in hereditary hemorrhagic telangiectasia.

Hereditary hemorrhagic telangiectasia (HHT), or Rendu-Osler-Weber disease, is an autosomal dominant disorder of localized angiodysplasia, although it is sometimes mistakenly identified as a hemostatic disorder due to its associated characteristic bleeding. The vascular lesions that develop consist of direct arteriovenous connections without an intervening capillary bed. Germline mutations in one of two different genes, endoglin or ALK-1, can cause HHT. Both are members of the transforming growth factor (TGF)-beta receptor family of proteins, and are expressed primarily on the surface of endothelial cells. They are associated together in a receptor complex on the cell surface. Biochemical studies suggest that endoglin modulates TGF-beta signaling through ALK-1 and the type I TGF-beta receptor. Most mutations identified in endoglin and ALK-1 create null alleles, which lead to reduced message or protein levels. A model of haploinsufficiency is proposed, in which inheritance of a mutation predisposes an individual to develop HHT-associated vascular lesions. The factors that initiate lesion formation are unknown, but disruption of these genes in mice should provide animal models to address these and other important questions about the pathogenesis of HHT.

Authors
Marchuk, DA
MLA Citation
Marchuk, DA. "Genetic abnormalities in hereditary hemorrhagic telangiectasia." Curr Opin Hematol 5.5 (September 1998): 332-338. (Review)
PMID
9776212
Source
pubmed
Published In
Current Opinion in Hematology
Volume
5
Issue
5
Publish Date
1998
Start Page
332
End Page
338

Familial segregation of hemangiomas and vascular malformations as an autosomal dominant trait.

BACKGROUND: The pathogenesis of infantile hemangiomas is not yet understood. Growth factors and hormonal and mechanical influences have been thought to affect the focal abnormal growth of endothelial cells in these lesions. However, these influences may represent secondary responses to an underlying primary molecular event leading to the development of hemangiomas. OBSERVATIONS: We report the rare familial occurrence of hemangiomas and/or vascular malformations in 6 kindreds, suggesting autosomal dominant inheritance. In these families, multiple generations (2-4) were affected by hemangiomas or vascular malformations. In contrast to the generally accepted female-male ratio of 3:1 to 4:1 associated with sporadic hemangiomas, the families with hemangiomas in our study demonstrated a 2:1 ratio. Additionally, vascular malformations and hemangiomas were present in different members of the same family. The vascular lesions appeared to be transmitted in an autosomal dominant fashion with moderate to high penetrance. CONCLUSIONS: We have identified 6 families demonstrating autosomal dominant segregation of childhood hemangiomas. Additionally, family members with vascular malformations were identified in these kindreds. Physicians caring for children with hemangiomas and vascular malformations should include in their medical histories inquiries about vascular lesions in other family members, even when obvious lesions are not present in the parents. The identification of the mutation(s) underlying vascular lesions will provide insight into the pathogenesis of these familial hemangiomas and, potentially, common sporadic hemangiomas. In addition, such research would shed light on the regulation of angiogenic processes during development.

Authors
Blei, F; Walter, J; Orlow, SJ; Marchuk, DA
MLA Citation
Blei, F, Walter, J, Orlow, SJ, and Marchuk, DA. "Familial segregation of hemangiomas and vascular malformations as an autosomal dominant trait." Arch Dermatol 134.6 (June 1998): 718-722.
PMID
9645641
Source
pubmed
Published In
Archives of Dermatology
Volume
134
Issue
6
Publish Date
1998
Start Page
718
End Page
722

Report on the workshop on Hereditary Hemorrhagic Telangiectasia, July 10-11, 1997.

Authors
Marchuk, DA; Guttmacher, AE; Penner, JA; Ganguly, P
MLA Citation
Marchuk, DA, Guttmacher, AE, Penner, JA, and Ganguly, P. "Report on the workshop on Hereditary Hemorrhagic Telangiectasia, July 10-11, 1997." Am J Med Genet 76.3 (March 19, 1998): 269-273.
PMID
9508248
Source
pubmed
Published In
American Journal of Medical Genetics Part A
Volume
76
Issue
3
Publish Date
1998
Start Page
269
End Page
273

Quantitative DNA pooling to increase the efficiency of linkage analysis in autosomal dominant disease.

DNA pooling is an efficient method to rapidly perform genome-wide linkage scans in autosomal recessive diseases in inbred populations where affected individuals are likely to be homozygous for alleles near the disease gene locus. We wanted to examine whether this approach would detect linkage in autosomal dominant (AD) disorders where affected individuals may share one allele identical by descent at loci tightly linked to the disease. Two large outbred pedigrees in which the AD diseases familial venous malformation (FVM) and hereditary hemorrhagic telangiectasia (HHT1), linked to 9p and 9q, respectively, were investigated. Separate pools of DNA from affected (n = 21 for FVM and 17 for HHT1) and unaffected family members (n = 9 FVM and HHT1), and 25 unrelated population controls were established. Polymorphic markers spanning chromosome 9 at approximately 13.5-cM intervals were amplified using standard PCR. Allele quantitation was performed with a fluorimager. Visual inspection of allele intensities and frequency distributions suggested a shift in frequency of the most common allele in the affecteds lane when compared to control lanes for markers within 30 cM of the FVM and HHT1 loci. These subjective assessments were confirmed statistically by testing for the difference between two proportions (one-sided; P < or = 0.05). When using population controls, the true-positive rates for FVM and HHT1 were 5/5 and 2/5 markers, respectively. False-positive rates for FVM and HHT1 were 3/9 and 2/9, respectively. In both AD diseases investigated, quantitative DNA pooling detected shifts in allele frequency, thus identifying areas of known linkage in most cases. The utility of this technique depends on the size of the pedigree, frequency of the disease-associated allele in the population, and the choice of appropriate controls. Although the false-positive rate appears to be high, this approach still serves to reduce the amount of overall genotyping by about 60%. DNA pooling merits further investigation as a potential strategy in increasing the efficiency of genomic linkage scans.

Authors
Damji, KF; Gallione, CJ; Allingham, RR; Slotterbeck, B; Guttmacher, AE; Pasyk, KA; Vance, JM; Pericak-Vance, MA; Speer, MC; Marchuk, DA
MLA Citation
Damji, KF, Gallione, CJ, Allingham, RR, Slotterbeck, B, Guttmacher, AE, Pasyk, KA, Vance, JM, Pericak-Vance, MA, Speer, MC, and Marchuk, DA. "Quantitative DNA pooling to increase the efficiency of linkage analysis in autosomal dominant disease." Hum Genet 102.2 (February 1998): 207-212.
PMID
9521591
Source
pubmed
Published In
Human Genetics
Volume
102
Issue
2
Publish Date
1998
Start Page
207
End Page
212

Novel missense and frameshift mutations in the activin receptor-like kinase-1 gene in hereditary hemorrhagic telangiectasia. Mutations in brief no. 164. Online.

Hereditary hemmorrhagic telangiectasia (HHT) is an autosomal dominant disorder characterized by multisystemic vascular dyplasia and recurrent hemorrhage. One of the causative genes is the activin receptor-like kinase-1 (ALK-1) gene located on chromosome 12q13. ALK-1 is an endothelial cell type I receptor for the TGF-beta superfamily of ligands. As a number of mutations have been identified in the kinase domain of ALK-1, we initiated a mutation analysis specifically targeting the first four coding exons of ALK-1 in order to determine if mutations in the extracellular and transmembrane domains are also present in HHT. Six new mutations have been identified. Three frameshift mutations were identified in exons encoding the extracellular and transmembrane domains. These mutations would grossly truncate the ALK-1 protein and are thus classic null alleles. Three new missense mutations within the exons encoding the extracellular domain, in addition to two previously described missense mutations, are located at or near highly conserved cysteines. These mutations may disrupt intra- or inter-molecular disulfide bridges required for ligand binding. The combined data suggest that both severe and subtle changes in the ALK-1 amino acid sequence can lead to receptor dysfunction and result in the HHT disease phenotype.

Authors
Klaus, DJ; Gallione, CJ; Anthony, K; Yeh, EY; Yu, J; Lux, A; Johnson, DW; Marchuk, DA
MLA Citation
Klaus, DJ, Gallione, CJ, Anthony, K, Yeh, EY, Yu, J, Lux, A, Johnson, DW, and Marchuk, DA. "Novel missense and frameshift mutations in the activin receptor-like kinase-1 gene in hereditary hemorrhagic telangiectasia. Mutations in brief no. 164. Online." Hum Mutat 12.2 (1998): 137-.
PMID
10694922
Source
pubmed
Published In
Human Mutation
Volume
12
Issue
2
Publish Date
1998
Start Page
137
DOI
10.1002/(SICI)1098-1004(1998)12:2<137::AID-HUMU16>3.0.CO;2-J

Mutation and expression analysis of the endoglin gene in hereditary hemorrhagic telangiectasia reveals null alleles.

Hereditary Hemorrhagic Telangiectasia (HHT) is an autosomal dominant disorder characterized by multisystemic vascular dysplasia and recurrent hemorrhage from the sites of vascular lesions. Two genes have been identified for HHT. Endoglin, a TGF-beta binding protein which maps to chromosome 9q3, is the gene for HHT1. The type and location of most of the previously described mutations in the endoglin (ENG) gene suggested a dominant-negative model of receptor-complex dysfunction for the molecular basis of this disorder. In this article we describe 11 novel ENG mutations in HHT kindreds, which include missense and splice-site mutations. Two identical missense mutations in unrelated families disrupt the start codon of the gene. In addition, some frameshift and nonsense mutations lead to very low or undetectable levels of transcript from the mutant allele. These combined data suggest that the nature of most ENG mutations is to create a null (nonfunctional) allele, and that there is no requirement for the synthesis of a truncated endoglin protein in the pathogenesis of HHT.

Authors
Gallione, CJ; Klaus, DJ; Yeh, EY; Stenzel, TT; Xue, Y; Anthony, KB; McAllister, KA; Baldwin, MA; Berg, JN; Lux, A; Smith, JD; Vary, CP; Craigen, WJ; Westermann, CJ; Warner, ML; Miller, YE; Jackson, CE; Guttmacher, AE; Marchuk, DA
MLA Citation
Gallione, CJ, Klaus, DJ, Yeh, EY, Stenzel, TT, Xue, Y, Anthony, KB, McAllister, KA, Baldwin, MA, Berg, JN, Lux, A, Smith, JD, Vary, CP, Craigen, WJ, Westermann, CJ, Warner, ML, Miller, YE, Jackson, CE, Guttmacher, AE, and Marchuk, DA. "Mutation and expression analysis of the endoglin gene in hereditary hemorrhagic telangiectasia reveals null alleles." Hum Mutat 11.4 (1998): 286-294.
PMID
9554745
Source
pubmed
Published In
Human Mutation
Volume
11
Issue
4
Publish Date
1998
Start Page
286
End Page
294
DOI
10.1002/(SICI)1098-1004(1998)11:4<286::AID-HUMU6>3.0.CO;2-B

Erratum: Familial segregation of hemangiomas and vascular malformations as an autosomal dominant trait (Archives of Dermatology (June 1998) 134 (718- 722))

Authors
Blei, F; Walter, J; Orlow, SJ; Marchuk, DA
MLA Citation
Blei, F, Walter, J, Orlow, SJ, and Marchuk, DA. "Erratum: Familial segregation of hemangiomas and vascular malformations as an autosomal dominant trait (Archives of Dermatology (June 1998) 134 (718- 722))." Archives of Dermatology 134.11 (1998): 1425--.
Source
scival
Published In
Archives of Dermatology
Volume
134
Issue
11
Publish Date
1998
Start Page
1425-

The activin receptor-like kinase 1 gene: genomic structure and mutations in hereditary hemorrhagic telangiectasia type 2.

The activin receptor-like kinase 1 gene (ALK-1) is the second locus for the autosomal dominant vascular disease hereditary hemorrhagic telangiectasia (HHT). In this paper we present the genomic structure of the ALK-1 gene, a type I serine-threonine kinase receptor expressed predominantly in endothelial cells. The coding region is contained within nine exons, spanning < 15 kb of genomic DNA. All introns follow the GT-AG rule, except for intron 6, which has a TAG/gcaag 5' splice junction. The positions of introns in the intracellular domain are almost identical to those of the mouse serine-threonine kinase receptor TSK-7L. By sequencing ALK-1 from genomic DNA, mutations were found in six of six families with HHT either shown to link to chromosome 12q13 or in which linkage of HHT to chromosome 9q33 had been excluded. Mutations were also found in three of six patients from families in which available linkage data were insufficient to allow certainty with regard to the locus involved. The high rate of detection of mutations by genomic sequencing of ALK-1 suggests that this will be a useful diagnostic test for HHT2, particularly where preliminary linkage to chromosome 12q13 can be established. In two cases in which premature termination codons were found in genomic DNA, the mutant mRNA was either not present or present at barely detectable levels. These data suggest that mutations in ALK-1 are functionally null alleles.

Authors
Berg, JN; Gallione, CJ; Stenzel, TT; Johnson, DW; Allen, WP; Schwartz, CE; Jackson, CE; Porteous, ME; Marchuk, DA
MLA Citation
Berg, JN, Gallione, CJ, Stenzel, TT, Johnson, DW, Allen, WP, Schwartz, CE, Jackson, CE, Porteous, ME, and Marchuk, DA. "The activin receptor-like kinase 1 gene: genomic structure and mutations in hereditary hemorrhagic telangiectasia type 2." Am J Hum Genet 61.1 (July 1997): 60-67.
PMID
9245985
Source
pubmed
Published In
The American Journal of Human Genetics
Volume
61
Issue
1
Publish Date
1997
Start Page
60
End Page
67
DOI
10.1086/513903

Endoglin gene polymorphism as a risk factor for sporadic intracerebral hemorrhage.

Intracerebral hemorrhage (ICH) is a common and serious type of stroke. Recent studies have shown that inherited factors that affect the development of the vessel wall can increase the risk of ICH. We studied endoglin as a candidate gene in patients with sporadic ICH, since mutations in this gene can cause telangiectasia formation. One hundred three patients with sporadic ICH and 202 controls were studied. The polymerase chain reaction and single-strand conformational polymorphism analysis were used to screen for mutations in exon 7 of the endoglin gene. No coding mutations in exon 7 were identified in the ICH patients or controls. A 6-base intronic insertion was found 26 bases beyond the 3' end of exon 7. The homozygous form of the insertion was present in 9 of 103 (8.7%) ICH patients compared with 4 of 202 (2.0%) controls, p = 0.012 (odds ratio 4.8 [95% confidence interval, 1.28, 21.60]). Analysis of the endoglin transcript around the insertion did not reveal any changes in the RNA sequence. There were no obvious clinical features that distinguished the ICH patients with the homozygous insertion from the other patients. The pathophysiologic mechanism underlying this association remains to be determined.

Authors
Alberts, MJ; Davis, JP; Graffagnino, C; McClenny, C; Delong, D; Granger, C; Herbstreith, MH; Boteva, K; Marchuk, DA; Roses, AD
MLA Citation
Alberts, MJ, Davis, JP, Graffagnino, C, McClenny, C, Delong, D, Granger, C, Herbstreith, MH, Boteva, K, Marchuk, DA, and Roses, AD. "Endoglin gene polymorphism as a risk factor for sporadic intracerebral hemorrhage." Ann Neurol 41.5 (May 1997): 683-686.
PMID
9153532
Source
pubmed
Published In
Annals of Neurology
Volume
41
Issue
5
Publish Date
1997
Start Page
683
End Page
686
DOI
10.1002/ana.410410519

The molecular genetics of hereditary hemorrhagic telangiectasia

Authors
Marchuk, DA
MLA Citation
Marchuk, DA. "The molecular genetics of hereditary hemorrhagic telangiectasia." Chest 111.6 SUPPL. (1997): 79S-82S.
Source
scival
Published In
Chest
Volume
111
Issue
6 SUPPL.
Publish Date
1997
Start Page
79S
End Page
82S

Vascular dysmorphogenesis caused by an activating mutation in the receptor tyrosine kinase TIE2.

Venous malformations (VMs), the most common errors of vascular morphogenesis in humans, are composed of dilated, serpiginous channels. The walls of the channels have a variable thickness of smooth muscle; some mural regions lack smooth muscle altogether. A missense mutation resulting in an arginine-to-tryptophan substitution at position 849 in the kinase domain of the receptor tyrosine kinase TIE2 segregates with dominantly inherited VM in two unrelated families. Using proteins expressed in insect cells, we demonstrate that the mutation results in increased activity of TIE2. We conclude that an activating mutation in TIE2 causes inherited VMs in the two families and that the TIE2 signaling pathway is critical for endothelial cell-smooth muscle cell communication in venous morphogenesis.

Authors
Vikkula, M; Boon, LM; Carraway, KL; Calvert, JT; Diamonti, AJ; Goumnerov, B; Pasyk, KA; Marchuk, DA; Warman, ML; Cantley, LC; Mulliken, JB; Olsen, BR
MLA Citation
Vikkula, M, Boon, LM, Carraway, KL, Calvert, JT, Diamonti, AJ, Goumnerov, B, Pasyk, KA, Marchuk, DA, Warman, ML, Cantley, LC, Mulliken, JB, and Olsen, BR. "Vascular dysmorphogenesis caused by an activating mutation in the receptor tyrosine kinase TIE2." Cell 87.7 (December 27, 1996): 1181-1190.
PMID
8980225
Source
pubmed
Published In
Cell
Volume
87
Issue
7
Publish Date
1996
Start Page
1181
End Page
1190

Mutations in the activin receptor-like kinase 1 gene in hereditary haemorrhagic telangiectasia type 2.

Hereditary haemorrhagic telangiectasia, or Osler-Rendu-Weber (ORW) syndrome, is an autosomal dominant vascular dysplasia. So far, two loci have been demonstrated for ORW. Linkage studies established an ORW locus at chromosome 9q3; endoglin was subsequently identified as the ORW1 gene. A second locus, designated ORW2, was mapped to chromosome 12. Here we report a new 4 cM interval for ORW2 that does not overlap with any previously defined. A 1.38-Mb YAC contig spans the entire interval. It includes the activin receptor like kinase 1 gene (ACVRLK1 or ALK1), a member of the serine-threonine kinase receptor family expressed in endothelium. We report three mutations in the coding sequence of the ALK1 gene in those families which show linkage of the ORW phenotype to chromosome 12. Our data suggest a critical role for ALK1 in the control of blood vessel development or repair.

Authors
Johnson, DW; Berg, JN; Baldwin, MA; Gallione, CJ; Marondel, I; Yoon, SJ; Stenzel, TT; Speer, M; Pericak-Vance, MA; Diamond, A; Guttmacher, AE; Jackson, CE; Attisano, L; Kucherlapati, R; Porteous, ME; Marchuk, DA
MLA Citation
Johnson, DW, Berg, JN, Baldwin, MA, Gallione, CJ, Marondel, I, Yoon, SJ, Stenzel, TT, Speer, M, Pericak-Vance, MA, Diamond, A, Guttmacher, AE, Jackson, CE, Attisano, L, Kucherlapati, R, Porteous, ME, and Marchuk, DA. "Mutations in the activin receptor-like kinase 1 gene in hereditary haemorrhagic telangiectasia type 2." Nat Genet 13.2 (June 1996): 189-195.
PMID
8640225
Source
pubmed
Published In
Nature Genetics
Volume
13
Issue
2
Publish Date
1996
Start Page
189
End Page
195
DOI
10.1038/ng0696-189

Chromosome breakpoint at 17q11.2 and insertion of DNA from three different chromosomes in a glioblastoma with exceptional glial fibrillary acidic protein expression.

A glioblastoma that retained glial fibrillary acidic protein (GFAP) in culture has a break in the long arm of chromosome 17 at band 17q11.2. DNA inserted at this breakpoint came from chromosome bands 3p21, 3q23, 16q11.2, and 22q11.2. These chromosome fragments were inserted in band 17q11.2 proximal to the neurofibromatosis-1 (NF-1) gene and neu (HER2; erbB2) oncogene loci. The glioblastoma also contained a reciprocal translocation between 16p12 and 20p12. These structural abnormalities, previously undescribed in gliomas, were demonstrated by high-resolution chromosome banding, microdissection, and fluorescence in situ hybridization (FISH). Numerical changes typical of glioblastoma were present: gain of chromosome 7 and losses of chromosomes 10, 13, and 22. The complex chromosome origin of DNA inserted in this glioma chromosome is described. The association of two infrequent events in this single glioblastoma line, this complex insertion and retention of GFAP expression, is not likely to be a chance occurrence. It raises the possibility of an association between the two events.

Authors
McKeever, PE; Dennis, TR; Burgess, AC; Meltzer, PS; Marchuk, DA; Trent, JM
MLA Citation
McKeever, PE, Dennis, TR, Burgess, AC, Meltzer, PS, Marchuk, DA, and Trent, JM. "Chromosome breakpoint at 17q11.2 and insertion of DNA from three different chromosomes in a glioblastoma with exceptional glial fibrillary acidic protein expression." Cancer Genet Cytogenet 87.1 (March 1996): 41-47.
PMID
8646740
Source
pubmed
Published In
Cancer Genetics and Cytogenetics
Volume
87
Issue
1
Publish Date
1996
Start Page
41
End Page
47

Clinical heterogeneity in hereditary haemorrhagic telangiectasia: are pulmonary arteriovenous malformations more common in families linked to endoglin?

Pulmonary arteriovenous malformations (PAVMs) occur in up to 27% of patients with hereditary haemorrhagic telangiectasia (HHT) and are associated with a rate of paradoxical cerebral embolism at presentation of up to 36%. At least two different loci have been shown for HHT. Mutations in endoglin have been found in some families and the locus designated ORW1. In other families this locus has been excluded. In this paper we confirm that in families linked to ORW1 there is a prevalence of PAVMs among affected members of 29.2%, compared to a prevalence of 2.9% in families in which this locus has been excluded (chi 2 = 19.2, p < 0.001). This information can be used to decide how to screen HHT patients for PAVMs.

Authors
Berg, JN; Guttmacher, AE; Marchuk, DA; Porteous, ME
MLA Citation
Berg, JN, Guttmacher, AE, Marchuk, DA, and Porteous, ME. "Clinical heterogeneity in hereditary haemorrhagic telangiectasia: are pulmonary arteriovenous malformations more common in families linked to endoglin?." J Med Genet 33.3 (March 1996): 256-257.
PMID
8728706
Source
pubmed
Published In
Journal of medical genetics
Volume
33
Issue
3
Publish Date
1996
Start Page
256
End Page
257

Hereditary hemorrhagic telangiectasia [2]

Authors
Shovlin, CL; Hughes, JMB; Kjeldsen, AD; Vase, P; Oxhoj, H; Guttmacher, AE; Marchuk, DA; Jr, RLW
MLA Citation
Shovlin, CL, Hughes, JMB, Kjeldsen, AD, Vase, P, Oxhoj, H, Guttmacher, AE, Marchuk, DA, and Jr, RLW. "Hereditary hemorrhagic telangiectasia [2]." New England Journal of Medicine 334.5 (1996): 330-331.
Source
scival
Published In
New England Journal of Medicine
Volume
334
Issue
5
Publish Date
1996
Start Page
330
End Page
331
DOI
10.1056/NEJM199602013340514

Identification and mapping of type 1 neurofibromatosis (NF1) homologous loci.

During the establishment of a YAC contig for the type 1 neurofibromatosis (NF1) region on human chromosome 17q11.2, several YAC clones were isolated which originated from a different chromosome but which retained strong homology to NF1 coding regions (Marchuk et al., 1992). Fluorescence in situ hybridization (FISH) using these clones has identified NF1-homologous loci on several human chromosomes, including 2, 14, 15, 21, and 22. PCR amplification using primers originally designed to amplify NF1 exons has confirmed these chromosome localizations and, in addition, has revealed NF1-homologous sequences on chromosomes 12 and 20. Sequence analysis of the amplified products has demonstrated that (1) most of these loci have > 90% identity with NF1 sequences; (2) most of these loci represent nonprocessed pseudogenes; and (3) for the chromosome 12 locus, the two described regions of homology with NF1 have open reading frames. Finally, we have identified two novel alpha-satellite DNA repeat units in proximity to NF1-homologous sequences on chromosome 14. Their association suggests a mechanism for dispersion of the NF1-homologous loci based on alpha-satellite-mediated exchange between nonhomologous chromosomes.

Authors
Cummings, LM; Trent, JM; Marchuk, DA
MLA Citation
Cummings, LM, Trent, JM, and Marchuk, DA. "Identification and mapping of type 1 neurofibromatosis (NF1) homologous loci." Cytogenet Cell Genet 73.4 (1996): 334-340.
PMID
8751390
Source
pubmed
Published In
Cytogenetics and cell genetics
Volume
73
Issue
4
Publish Date
1996
Start Page
334
End Page
340

Refined localization of the cerebral cavernous malformation gene (CCM1) to a 4-cM interval of chromosome 7q contained in a well-defined YAC contig.

Cerebral cavernous malformations (CCM) are vascular lesions present in some 20 million people worldwide that are responsible for seizures, migraine, hemorrhage, and other neurologic problems. Familial cases ofCCM can be inherited as an autosomal dominant disorder with variable expression. A gene for CCM (CCM/)was recently mapped to a 33-cM segment of chromosome 7q in a large Hispanic family (Dubovsky et al.1995). Here, the collection of several new short tandem repeat polymorphisms (STRPs) within the region of interest on 7q and the refinement of the marker order in this region using both linkage analysis in CEPH families and especially YAC-based STS content mapping are described. Affected members of three Hispanic families share allele haplotypes indicating a common ancestral mutation within these families. Using the shared haplotype information along with analysis of crossovers in affected individuals from both the Hispanic and Caucasian families, the region likely to contain the CCMI gene has been reduced to a 4-cM segment of 7q between D7S2410 and D7S689. All markers within the refined chromosomal segment were located on a single YAC contig estimated to be approximately 2 Mb in size. Four potential candidate genes have been mapped to this region.

Authors
Johnson, EW; Iyer, LM; Rich, SS; Orr, HT; Gil-Nagel, A; Kurth, JH; Zabramski, JM; Marchuk, DA; Weissenbach, J; Clericuzio, CL; Davis, LE; Hart, BL; Gusella, JF; Kosofsky, BE; Louis, DN; Morrison, LA; Green, ED; Weber, JL
MLA Citation
Johnson, EW, Iyer, LM, Rich, SS, Orr, HT, Gil-Nagel, A, Kurth, JH, Zabramski, JM, Marchuk, DA, Weissenbach, J, Clericuzio, CL, Davis, LE, Hart, BL, Gusella, JF, Kosofsky, BE, Louis, DN, Morrison, LA, Green, ED, and Weber, JL. "Refined localization of the cerebral cavernous malformation gene (CCM1) to a 4-cM interval of chromosome 7q contained in a well-defined YAC contig." Genome Res 5.4 (November 1995): 368-380.
PMID
8750196
Source
pubmed
Published In
Genome research
Volume
5
Issue
4
Publish Date
1995
Start Page
368
End Page
380

Hereditary hemorrhagic telangiectasia.

Authors
Guttmacher, AE; Marchuk, DA; White, RI
MLA Citation
Guttmacher, AE, Marchuk, DA, and White, RI. "Hereditary hemorrhagic telangiectasia." N Engl J Med 333.14 (October 5, 1995): 918-924. (Review)
PMID
7666879
Source
pubmed
Published In
The New England journal of medicine
Volume
333
Issue
14
Publish Date
1995
Start Page
918
End Page
924
DOI
10.1056/NEJM199510053331407

Six novel mutations in the endoglin gene in hereditary hemorrhagic telangiectasia type 1 suggest a dominant-negative effect of receptor function.

Authors
McAllister, KA; Baldwin, MA; Thukkani, AK; Gallione, CJ; Berg, JN; Porteous, ME; Guttmacher, AE; Marchuk, DA
MLA Citation
McAllister, KA, Baldwin, MA, Thukkani, AK, Gallione, CJ, Berg, JN, Porteous, ME, Guttmacher, AE, and Marchuk, DA. "Six novel mutations in the endoglin gene in hereditary hemorrhagic telangiectasia type 1 suggest a dominant-negative effect of receptor function." Hum Mol Genet 4.10 (October 1995): 1983-1985.
PMID
8595426
Source
pubmed
Published In
Human Molecular Genetics
Volume
4
Issue
10
Publish Date
1995
Start Page
1983
End Page
1985

A second locus for hereditary hemorrhagic telangiectasia maps to chromosome 12.

Hereditary hemorrhagic telangiectasia (HHT) or Osler-Rendu-Weber (ORW) disease is an autosomal dominant vascular dysplasia. Initial linkage studies identified an ORW gene localized to 9q33-q34 but with some families clearly excluding this region. A probable correlation in clinical phenotype between the 9q3-linked families and unlinked families was described with a significantly lower incidence of pulmonary arteriovenous malformations observed in the unlinked families. In this study we examined four unrelated ORW families for which linkage to chromosome 9q33-q34 has been previously excluded. Linkage was established for all four families to markers on chromosome 12, with a combined maximum lod score of 10.77 (theta = 0.04) with D12S339. Mapping of crossovers using haplotype analysis indicated that the candidate region lies in an 11-CM interval between D12S345 and D12S339, in the pericentromeric region of chromosome 12. A map location for a second ORW locus is thus established that exhibits a significantly reduced incidence of pulmonary involvement.

Authors
Johnson, DW; Berg, JN; Gallione, CJ; McAllister, KA; Warner, JP; Helmbold, EA; Markel, DS; Jackson, CE; Porteous, ME; Marchuk, DA
MLA Citation
Johnson, DW, Berg, JN, Gallione, CJ, McAllister, KA, Warner, JP, Helmbold, EA, Markel, DS, Jackson, CE, Porteous, ME, and Marchuk, DA. "A second locus for hereditary hemorrhagic telangiectasia maps to chromosome 12." Genome Res 5.1 (August 1995): 21-28.
PMID
8717052
Source
pubmed
Published In
Genome research
Volume
5
Issue
1
Publish Date
1995
Start Page
21
End Page
28

A locus for cerebral cavernous malformations maps to chromosome 7q in two families.

Cavernous malformations (angiomas) affecting the central nervous system and retina can be inherited in autosomal dominant pattern (OMIM 116860). These vascular lesions may remain clinically silent or lead to a number of neurological symptoms including seizure, intracranial hemorrhage, focal neurological deficit, and migraine. We have mapped a gene for this disorder in two families, one of Italian-American origin and one of Mexican-American origin, to markers on proximal 7q, with a combined maximum lod score of 3.92 (theta of zero) with marker D7S479. Haplotype analysis of these families places the locus between markers D7S502 proximally and D7S515 distally, an interval of approximately 41 cM. The location distinguishes this disorder from an autosomal dominant vascular malformation syndrome where lesions are primarily cutaneous and that maps to 9p21.

Authors
Marchuk, DA; Gallione, CJ; Morrison, LA; Clericuzio, CL; Hart, BL; Kosofsky, BE; Louis, DN; Gusella, JF; Davis, LE; Prenger, VL
MLA Citation
Marchuk, DA, Gallione, CJ, Morrison, LA, Clericuzio, CL, Hart, BL, Kosofsky, BE, Louis, DN, Gusella, JF, Davis, LE, and Prenger, VL. "A locus for cerebral cavernous malformations maps to chromosome 7q in two families." Genomics 28.2 (July 20, 1995): 311-314.
PMID
8530042
Source
pubmed
Published In
Genomics
Volume
28
Issue
2
Publish Date
1995
Start Page
311
End Page
314

Assignment of human transforming growth factor-beta type I and type III receptor genes (TGFBR1 and TGFBR3) to 9q33-q34 and 1p32-p33, respectively.

Authors
Johnson, DW; Qumsiyeh, M; Benkhalifa, M; Marchuk, DA
MLA Citation
Johnson, DW, Qumsiyeh, M, Benkhalifa, M, and Marchuk, DA. "Assignment of human transforming growth factor-beta type I and type III receptor genes (TGFBR1 and TGFBR3) to 9q33-q34 and 1p32-p33, respectively." Genomics 28.2 (July 20, 1995): 356-357.
PMID
8530052
Source
pubmed
Published In
Genomics
Volume
28
Issue
2
Publish Date
1995
Start Page
356
End Page
357
DOI
10.1006/geno.1995.1157

COL5A1: fine genetic mapping and exclusion as candidate gene in families with nail-patella syndrome, tuberous sclerosis 1, hereditary hemorrhagic telangiectasia, and Ehlers-Danlos Syndrome type II.

COL5A1, the gene for the alpha 1 chain of type V collagen, has been considered a candidate gene for certain diseases based on chromosomal location and/or disease phenotype. We have employed 3'-untranslated region RFLPs to exclude COL5A1 as a candidate gene in families with tuberous sclerosis 1, Ehlers-Danlos syndrome type II, and nail-patella syndrome. In addition, we describe a polymorphic simple sequence repeat (SSR) within a COL5A1 intron. This SSR is used to exclude COL5A1 as a candidate gene in hereditary hemorrhagic telangiectasia (Osler-Rendu-Weber disease) and to add COL5A1 to the existing map of "index" markers of chromosome 9 by evaluation of the COL5A1 locus on the CEPH 40-family reference pedigree set. This genetic mapping places COL5A1 between markers D9S66 and D9S67.

Authors
Greenspan, DS; Northrup, H; Au, KS; McAllister, KA; Francomano, CA; Wenstrup, RJ; Marchuk, DA; Kwiatkowski, DJ
MLA Citation
Greenspan, DS, Northrup, H, Au, KS, McAllister, KA, Francomano, CA, Wenstrup, RJ, Marchuk, DA, and Kwiatkowski, DJ. "COL5A1: fine genetic mapping and exclusion as candidate gene in families with nail-patella syndrome, tuberous sclerosis 1, hereditary hemorrhagic telangiectasia, and Ehlers-Danlos Syndrome type II." Genomics 25.3 (February 10, 1995): 737-739.
PMID
7759113
Source
pubmed
Published In
Genomics
Volume
25
Issue
3
Publish Date
1995
Start Page
737
End Page
739

Somatic mutations in the neurofibromatosis 1 gene in gliomas and primitive neuroectodermal tumours

The increased frequency of glioma among neurofibromatosis 1 (NF1) patients suggests a general involvement of the NF1 gene in glioma tumourigenesis. Using the methodology of conventional Southern blotting with a complete panel of overlapping partial cDNAs covering the whole NF1 gene, we screened 31 gliomas of several different subtypes and 3 primitive neuroectodermal tumours (PNETs) from non-NF1 patients for aberrant restriction patterns in their tumour DNA samples. Clear evidence for somatic mutation events at the NF1 gene locus was found in 1 astrocytoma, 2 glioblastomas, 1 ependymoma and 1 PNET with an astrocytic component. These results suggest that the NF1 gene is important in suppressing tumours of neuroectodermal origin.

Authors
Thiel, G; Marczinek, K; Neumann, R; Witkowski, R; Marchuk, DA; Nurnberg, P
MLA Citation
Thiel, G, Marczinek, K, Neumann, R, Witkowski, R, Marchuk, DA, and Nurnberg, P. "Somatic mutations in the neurofibromatosis 1 gene in gliomas and primitive neuroectodermal tumours." Anticancer Research 15.6 B (1995): 2495-2499.
Source
scival
Published In
Anticancer research
Volume
15
Issue
6 B
Publish Date
1995
Start Page
2495
End Page
2499

Report on the Fourth International Workshop on Chromosome 9

The Fourth International Workshop on Chromosome 9 was a highly successful endeavor in terms of the growth of the map, both genetic and physical, the amount of data entered into GDB, and the continued comradeship in the sharing of data and resources that was exemplified. SIGMA remained a stable and valuable part of the chromosome 8 mapping effort. A new subsection outlining the morbid anatomy of chromosome 9 was included. Finally, specific goals were set for the community to aim for over the upcoming months. These included extending the information about the ease of use of genetic markers, and coordinating across numerous groups the meiotic breakpoint mapping of many microsatellite markers. Workshop files are available by anonymous ftp from ftp.gene.ucl.ac.uk (128.40.82.1) in the subdirectory /pub/c9workshop/1995, or by using a World Wide Web browser (such as Mosaic or Netscape) via the Chromosome 9 Home Page (at the URL http://www.gene.ucl.ac.uk/chr9home.html).

Authors
Pericak-Vance, MA; Bale, AE; Haines, JL; Kwiatkowski, DJ; Pilz, A; Slaugenhaupt, S; White, JA; Edwards, JH; Marchuk, D; Olopades, OI; Attwood, J; Povey, S
MLA Citation
Pericak-Vance, MA, Bale, AE, Haines, JL, Kwiatkowski, DJ, Pilz, A, Slaugenhaupt, S, White, JA, Edwards, JH, Marchuk, D, Olopades, OI, Attwood, J, and Povey, S. "Report on the Fourth International Workshop on Chromosome 9." Annals of Human Genetics 59.4 (1995): 347-365.
Source
scival
Published In
Annals of Human Genetics
Volume
59
Issue
4
Publish Date
1995
Start Page
347
End Page
365

A gene for familial venous malformations maps to chromosome 9p in a second large kindred

Venous malformations are a common form of vascular anomaly that cause pain and disfigurement and can be life threatening if they involve critical organs. They occur sporadically or in a familial form, where multiple lesions are usually present. We have identified a large kindred showing autosomal dominant inheritance of venous malformations. Using this family we confirm Linkage of a familial form of venous malformations to chromosome 9p. We suggest that blue rubber bleb naevus syndrome can be considered a particular manifestation of this form of familial venous malformations. The candidate region for this gene encompasses the interferon gene cluster and the MTS1 (p16) tumour suppressor gene.

Authors
Gallione, CJ; Pasyk, KA; Boon, LM; Lennon, F; Johnson, DW; Helmboid, EA; Markel, DS; Vikkula, M; Mulliken, JB; Warman, ML; Pericak-Vance, MA; Marchuk, DA
MLA Citation
Gallione, CJ, Pasyk, KA, Boon, LM, Lennon, F, Johnson, DW, Helmboid, EA, Markel, DS, Vikkula, M, Mulliken, JB, Warman, ML, Pericak-Vance, MA, and Marchuk, DA. "A gene for familial venous malformations maps to chromosome 9p in a second large kindred." Journal of Medical Genetics 32.3 (1995): 197-199.
PMID
7783168
Source
scival
Published In
Journal of Medical Genetics
Volume
32
Issue
3
Publish Date
1995
Start Page
197
End Page
199

Genetic heterogeneity in hereditary haemorrhagic telangiectasia: possible correlation with clinical phenotype.

Hereditary haemorrhagic telangiectasia (HHT) or Osler-Weber-Rendu syndrome is an autosomal dominant vascular dysplasia characterised by recurrent haemorrhage. Our initial linkage studies found an HHT gene to be localised to 9q3 in two large kindreds. In the present study, we examine an additional five unrelated HHT families. Linkage analysis in this region resulted in a peak multipoint location score of 13.03, 10 cM proximal of D9S60. We found significant evidence for heterogeneity of HHT. Multipoint analysis supports the family specific two point studies with odds of 3,000,000:1 showing linkage and heterogeneity over linkage and homogeneity. Four of the seven families give a posterior probability of > 99% of being of the linked type, and three families appear unlinked to this region of 9q, and by multipoint analysis completely exclude the candidate region for HHT. Two new crossovers in affected persons in one of the linked families further define the proximal border of the candidate region on 9q3. A possible correlation in clinical phenotype between the 9q3 linked families and unlinked families is described. Although six of the seven families clearly meet the clinical criteria for HHT diagnosis, a significant absence of pulmonary arteriovenous malformations is seen in all three 9q3 unlinked families. Genetic heterogeneity of HHT and its potential correlation with a clinical phenotype may have a significant impact on the clinical management and treatment of HHT patients.

Authors
McAllister, KA; Lennon, F; Bowles-Biesecker, B; McKinnon, WC; Helmbold, EA; Markel, DS; Jackson, CE; Guttmacher, AE; Pericak-Vance, MA; Marchuk, DA
MLA Citation
McAllister, KA, Lennon, F, Bowles-Biesecker, B, McKinnon, WC, Helmbold, EA, Markel, DS, Jackson, CE, Guttmacher, AE, Pericak-Vance, MA, and Marchuk, DA. "Genetic heterogeneity in hereditary haemorrhagic telangiectasia: possible correlation with clinical phenotype." J Med Genet 31.12 (December 1994): 927-932.
PMID
7891374
Source
pubmed
Published In
Journal of medical genetics
Volume
31
Issue
12
Publish Date
1994
Start Page
927
End Page
932

A disease locus for hereditary haemorrhagic telangiectasia maps to chromosome 9q33-34.

Hereditary haemorrhagic telangiectasia (HHT), or Osler-Weber-Rendu disease, is an autosomal dominant vascular dysplasia of unknown pathogenesis leading to 'widespread' dermal, mucosal and visceral telangiectases and recurrent haemorrhage. We have mapped the HHT gene, by linkage analysis, to markers on 9q33-34 in two large multi-generation families. Haplotype analysis and mapping of recombination breakpoints gives a 4 cM interval between D9S61 and D9S63 as the most likely location of the gene. The closest marker, D9S65, is estimated to be within 1 cM of the gene and shows a combined lod score of 11.41. Two potential candidate genes, COL5A1 and ZNF79, are also located within 9q33-34. These results provide a starting point for the eventual cloning of the HHT gene.

Authors
McDonald, MT; Papenberg, KA; Ghosh, S; Glatfelter, AA; Biesecker, BB; Helmbold, EA; Markel, DS; Zolotor, A; McKinnon, WC; Vanderstoep, JL
MLA Citation
McDonald, MT, Papenberg, KA, Ghosh, S, Glatfelter, AA, Biesecker, BB, Helmbold, EA, Markel, DS, Zolotor, A, McKinnon, WC, and Vanderstoep, JL. "A disease locus for hereditary haemorrhagic telangiectasia maps to chromosome 9q33-34." Nat Genet 6.2 (February 1994): 197-204.
PMID
8162075
Source
pubmed
Published In
Nature Genetics
Volume
6
Issue
2
Publish Date
1994
Start Page
197
End Page
204
DOI
10.1038/ng0294-197

Endoglin, a TGF-β binding protein of endothelial cells, is the gene for hereditary haemorrhagic telangiectasia type 1

Hereditary haemorrhagic telangiectasia (HHT) is an autosomal dominant disorder characterized by multisystemic vascular dysplasia and recurrent haemorrhage. Linkage for some families has been established to chromosome 9q33-q34. In the present study, endoglin, a transforming growth factor β (TGF-β) binding protein, was analysed as a candidate gene for the disorder based on chromosomal location, expression pattern and function. We have identified mutations in three affected individuals: a C to G substitution converting a tyrosine to a termination codon, a 39 base pair deletion and a 2 basepair deletion which creates a premature termination codon. We have identified endoglin as the HHT gene mapping to 9q3 and have established HHT as the first human disease defined by a mutation in a member of the TGF-β receptor complex.

Authors
McAllister, KA; Grogg, KM; Johnson, DW; Gallione, CJ; Baldwin, MA; Jackson, CE; Helmbold, EA; Markel, DS; McKinnon, WC; Murrell, J; McCormick, MK; Pericak-Vance, MA; Heutink, P; Oostra, BA; Haitjema, T; Westerman, CJJ; Porteous, ME; Guttmacher, AE; Letarte, M; Marchuk, DA
MLA Citation
McAllister, KA, Grogg, KM, Johnson, DW, Gallione, CJ, Baldwin, MA, Jackson, CE, Helmbold, EA, Markel, DS, McKinnon, WC, Murrell, J, McCormick, MK, Pericak-Vance, MA, Heutink, P, Oostra, BA, Haitjema, T, Westerman, CJJ, Porteous, ME, Guttmacher, AE, Letarte, M, and Marchuk, DA. "Endoglin, a TGF-β binding protein of endothelial cells, is the gene for hereditary haemorrhagic telangiectasia type 1." Nature Genetics 8.4 (1994): 345-351.
PMID
7894484
Source
scival
Published In
Nature Genetics
Volume
8
Issue
4
Publish Date
1994
Start Page
345
End Page
351
DOI
10.1038/ng1294-345

Genetic heterogeneity in hereditary haemorrhagic telangiectasia

A locus causing hereditary haemorrhagic telangiectasia (HHT) has recently been mapped to 9q34 in four families and designated HHT1. In this paper, the results of a linkage study showing genetic heterogeneity in four families in whom HHT is segregating are reported. All the previously reported 9q34 linked families contain at least one affected member with a symptomatic pulmonary arteriovenous malformation. We postulate that clinical heterogeneity may also be a feature of HHT with a significantly higher predisposition to symptomatic PAVMs associated with the HHT1 linked families.

Authors
Porteous, MEM; Curtis, A; Williams, O; Marchuk, D; Bhattacharya, SS; Burn, J
MLA Citation
Porteous, MEM, Curtis, A, Williams, O, Marchuk, D, Bhattacharya, SS, and Burn, J. "Genetic heterogeneity in hereditary haemorrhagic telangiectasia." Journal of Medical Genetics 31.12 (1994): 925-926.
PMID
7891373
Source
scival
Published In
Journal of Medical Genetics
Volume
31
Issue
12
Publish Date
1994
Start Page
925
End Page
926

An EcoRI RFLP in the 5' region of the human NF1 gene.

Von Recklinghausen neurofibromatosis or type 1 neurofibromatosis (NF1), is one of the most common autosomal dominant disorders. NF1 is characterized by neurofibromas, café-au-lait spots and Lisch nodules of the iris. The NF1 gene is located in 17q11.2. The restriction fragment length polymorphism reported here will be useful in linkage analysis in NF1 families.

Authors
Reyniers, E; De Boulle, K; Marchuk, DA; Andersen, LB; Collins, FS; Willems, PJ
MLA Citation
Reyniers, E, De Boulle, K, Marchuk, DA, Andersen, LB, Collins, FS, and Willems, PJ. "An EcoRI RFLP in the 5' region of the human NF1 gene." Hum Genet 92.6 (December 1993): 631-.
PMID
7903272
Source
pubmed
Published In
Human Genetics
Volume
92
Issue
6
Publish Date
1993
Start Page
631

A compound nucleotide repeat in the neurofibromatosis (NF1) gene.

Authors
Andersen, LB; Tarlé, SA; Marchuk, DA; Legius, E; Collins, FS
MLA Citation
Andersen, LB, Tarlé, SA, Marchuk, DA, Legius, E, and Collins, FS. "A compound nucleotide repeat in the neurofibromatosis (NF1) gene." Hum Mol Genet 2.7 (July 1993): 1083-.
PMID
8364559
Source
pubmed
Published In
Human Molecular Genetics
Volume
2
Issue
7
Publish Date
1993
Start Page
1083

Dinucleotide repeat polymorphism at the human erythropoietin receptor locus (EPOR) at 19p13.

Authors
McDonald, MT; Papenberg, KA; Glatfelter, AA; Vander-Stoep, JL; Marchuk, DA
MLA Citation
McDonald, MT, Papenberg, KA, Glatfelter, AA, Vander-Stoep, JL, and Marchuk, DA. "Dinucleotide repeat polymorphism at the human erythropoietin receptor locus (EPOR) at 19p13." Hum Mol Genet 2.5 (May 1993): 619-.
PMID
8518819
Source
pubmed
Published In
Human Molecular Genetics
Volume
2
Issue
5
Publish Date
1993
Start Page
619

Somatic deletion of the neurofibromatosis type 1 gene in a neurofibrosarcoma supports a tumour suppressor gene hypothesis.

Individuals with neurofibromatosis type 1 (NF1) have an increased risk of developing benign and malignant tumours. The NF1 gene is thought to be a tumour suppressor gene, yet no direct proof at the molecular level exists to support this hypothesis. Here we describe a neurofibrosarcoma from a patient with NF1 with loss of heterozygosity for all chromosome 17 polymorphisms tested. On the remaining chromosome 17 homologue, a 200 kilobase (kb) tumour specific deletion of NF1 was demonstrated. This is the first example of a homozygous inactivation of NF1 at the molecular level in a malignant tumour from an NF1 patient and the results strongly support the tumour suppressor gene hypothesis for this disease.

Authors
Legius, E; Marchuk, DA; Collins, FS; Glover, TW
MLA Citation
Legius, E, Marchuk, DA, Collins, FS, and Glover, TW. "Somatic deletion of the neurofibromatosis type 1 gene in a neurofibrosarcoma supports a tumour suppressor gene hypothesis." Nat Genet 3.2 (February 1993): 122-126.
PMID
8499945
Source
pubmed
Published In
Nature Genetics
Volume
3
Issue
2
Publish Date
1993
Start Page
122
End Page
126
DOI
10.1038/ng0293-122

A conserved alternative splice in the von Recklinghausen neurofibromatosis (NF1) gene produces two neurofibromin isoforms, both of which have GTPase-activating protein activity.

Sequence analysis has shown significant homology between the catalytic regions of the mammalian ras GTPase-activating protein (GAP), yeast Ira1p and Ira2p (inhibitory regulators of the RAS-cyclic AMP pathway), and neurofibromin, the protein encoded by the NF1 gene. Yeast expression experiments have confirmed that a 381-amino-acid segment of neurofibromin, dubbed the GAP-related domain (GRD), can function as a GAP. Using the RNA polymerase chain reaction with primers flanking the NF1-GRD, we have identified evidence for alternative splicing in this region of the NF1 gene. In addition to the already published sequence (type I), an alternative RNA carrying a 63-nucleotide insertion (type II) is present in all tissues examined, although the relative amounts of types I and II vary. The insertion is conserved across species but is not present in GAP, IRA1, or IRA2. GenBank searches have failed to identify significant similarity between the inserted sequence and known DNA or protein sequences, although the basic amino acid composition of the insertion shares features with nuclear targeting sequences. Expression studies in yeasts show that despite the partial disruption of the neurofibromin-IRA-GAP homology by this insertion, both forms of the NF1-GRD can complement loss of IRA function. In vivo assays designed to compare the GAP activity of the two alternatively spliced forms of the NF1-GRD show that both can increase the conversion of GTP-bound ras to its GDP-bound form, although the insertion of the 21 amino acids weakens this effect. The strong conservation of this alternative splicing suggests that both type I and II isoforms mediate important biological functions of neurofibromin.

Authors
Andersen, LB; Ballester, R; Marchuk, DA; Chang, E; Gutmann, DH; Saulino, AM; Camonis, J; Wigler, M; Collins, FS
MLA Citation
Andersen, LB, Ballester, R, Marchuk, DA, Chang, E, Gutmann, DH, Saulino, AM, Camonis, J, Wigler, M, and Collins, FS. "A conserved alternative splice in the von Recklinghausen neurofibromatosis (NF1) gene produces two neurofibromin isoforms, both of which have GTPase-activating protein activity." Mol Cell Biol 13.1 (January 1993): 487-495.
PMID
8417346
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
13
Issue
1
Publish Date
1993
Start Page
487
End Page
495

Analysis of the neurofibromatosis type 1 (NF1) GAP-related domain by site-directed mutagenesis

The gene for von Recklinghausen neurofibromatosis type 1 (NF1) was recently identified by positional cloning and found to encode a protein with sequence similarity to a family of eucaryotic GTPase-activating proteins (GAPs). Expression of the NF1-GAP-related domain (NF1GRD) has been shown to complement yeast strains deficient in the yeast GAP homologs, IRA1 and IRA2, to interact with human RAS proteins and to accelerate the conversion of ras-GTP to ras-GDP. Further analysis of this region has revealed a number of residues that are highly conserved between members of the GAP family. Mutational analysis of a representative number of these residues produced one of three effects: (1) no change in NF1GRD function, (2) complete disruption of NF1GRD function and (3) intermediate retention of NF1GRD function. One of these mutations at residue 1423 was shown to have reduced ability to negatively regulate ras in yeast, which is interesting in light of a recent report demonstrating a similar naturally occurring mutation in human malignancies.

Authors
Gutmann, DH; Boguski, M; Marchuk, D; Wigler, M; Collins, FS; Ballester, R
MLA Citation
Gutmann, DH, Boguski, M, Marchuk, D, Wigler, M, Collins, FS, and Ballester, R. "Analysis of the neurofibromatosis type 1 (NF1) GAP-related domain by site-directed mutagenesis." Oncogene 8.3 (1993): 761-769.
PMID
8437860
Source
scival
Published In
Oncogene
Volume
8
Issue
3
Publish Date
1993
Start Page
761
End Page
769

The gene for a novel epidermal antigen maps near the neurofibromatosis 1 gene.

Recently the M17S1 gene, encoding an epidermal antigen thought to play a role in cell adhesion, was mapped to chromosome bands 17q11-q12, placing it in the vicinity of the gene for the genetic disorder neurofibromatosis 1 (NF1). The pleomorphic cutaneous lesions of NF1 and the precedent for other genes being embedded within the NF1 gene prompted us to investigate whether the M17S1 gene mapped near, or within, the NF1 gene. Genetic linkage analyses revealed that M17S1 was tightly linked to NF1 and mapped within the interval bounded by D17S58 and D17S54. Physical mapping of an M17S1 cDNA on somatic cell hybrids, yeast artificial chromosomes, and an NF1 patient with a deletion involving an entire NF1 allele demonstrated that M17S1 is located at least 180 kb centromeric to the NF1 gene. The distance between the genes suggests that M17S1 is unlikely to contribute to the NF1 phenotype since a gross chromosomal rearrangement would be required to disrupt expression of both genes.

Authors
Kayes, LM; Schroeder, WT; Marchuk, DA; Collins, FS; Riccardi, VM; Duvic, M; Stephens, K
MLA Citation
Kayes, LM, Schroeder, WT, Marchuk, DA, Collins, FS, Riccardi, VM, Duvic, M, and Stephens, K. "The gene for a novel epidermal antigen maps near the neurofibromatosis 1 gene." Genomics 14.2 (October 1992): 369-376.
PMID
1358802
Source
pubmed
Published In
Genomics
Volume
14
Issue
2
Publish Date
1992
Start Page
369
End Page
376

NF1-related locus on chromosome 15.

A neurofibromatosis type I (NF1)-related locus has been identified on chromosome 15. It contains a partial copy of the NF1 GAP-related domain, which is known to interact with the ras protooncogenes. However, the chromosome 15 sequence contains multiple deletions resulting in frameshift mutations and stop codons in several highly conserved sequence blocks. The locus on chromosome 15 therefore represents an NF1 pseudogene. This nonprocessed NF1 pseudogene may produce additional fragments in Southern blotting, pulsed-field gel, and PCR experiments with some NF1 cDNA probes or oligonucleotides. In addition, certain regions of the NF1 gene also cross-hybridize with a locus on chromosome 14. These loci must be considered in mutation analysis of patients with NF1 since aberrant findings may not always reflect changes in the NF1 gene.

Authors
Legius, E; Marchuk, DA; Hall, BK; Andersen, LB; Wallace, MR; Collins, FS; Glover, TW
MLA Citation
Legius, E, Marchuk, DA, Hall, BK, Andersen, LB, Wallace, MR, Collins, FS, and Glover, TW. "NF1-related locus on chromosome 15." Genomics 13.4 (August 1992): 1316-1318.
PMID
1505963
Source
pubmed
Published In
Genomics
Volume
13
Issue
4
Publish Date
1992
Start Page
1316
End Page
1318

A yeast artificial chromosome contig encompassing the type 1 neurofibromatosis gene.

The yeast artificial chromosome (YAC) system (Burke et al., 1987, Science 236: 806-812) allows the direct cloning of large regions of the genome. A YAC contig map of approximately 700 kb encompassing the region surrounding the type 1 neurofibromatosis (NF1) locus on 17q11.2 has been constructed. A single YAC containing the entire NF1 locus has been constructed by homologous recombination in yeast. In the process of contig construction a novel method of YAC end rescue has been developed by YAC circularization in yeast and plasmid rescue in bacteria. YACs containing homology to the NF1 region but mapping to another chromosome have also been discovered. Sequences of portions of the homologous locus indicate that this other locus is a nonprocessed pseudogene.

Authors
Marchuk, DA; Tavakkol, R; Wallace, MR; Brownstein, BH; Taillon-Miller, P; Fong, CT; Legius, E; Andersen, LB; Glover, TW; Collins, FS
MLA Citation
Marchuk, DA, Tavakkol, R, Wallace, MR, Brownstein, BH, Taillon-Miller, P, Fong, CT, Legius, E, Andersen, LB, Glover, TW, and Collins, FS. "A yeast artificial chromosome contig encompassing the type 1 neurofibromatosis gene." Genomics 13.3 (July 1992): 672-680.
PMID
1639394
Source
pubmed
Published In
Genomics
Volume
13
Issue
3
Publish Date
1992
Start Page
672
End Page
680

Sequencing and analysis of genomic fragments from the NF1 locus.

The sequence of five non-contiguous genomic fragments encompassing 14.4 kilobases from the NF1 locus have been determined by fluorescence-based automated DNA sequence analysis. These fragments included one kilobase of the NF1 coding region, which resulted in the identification of the intron/exon boundaries of five exons. Based on these sequences, five new NF1 exon-PCR assays have been developed, that could be useful for detecting new NF1 mutations. The genomic sequences were analyzed for the presence of Alu repetitive elements and their classification is described. This analysis may provide some insight into the characterization of genetic rearrangements resulting in disruption of the NF1 gene.

Authors
Martin-Gallardo, A; Marchuk, DA; Gocayne, J; Kerlavage, AR; McCombie, WR; Venter, JC; Collins, FS; Wallace, MR
MLA Citation
Martin-Gallardo, A, Marchuk, DA, Gocayne, J, Kerlavage, AR, McCombie, WR, Venter, JC, Collins, FS, and Wallace, MR. "Sequencing and analysis of genomic fragments from the NF1 locus." DNA Seq 3.4 (1992): 237-243.
PMID
1338369
Source
pubmed
Published In
DNA Sequence: Journal of DNA Mapping, Sequencing, and Analysis
Volume
3
Issue
4
Publish Date
1992
Start Page
237
End Page
243

cDNA cloning of the type 1 neurofibromatosis gene: complete sequence of the NF1 gene product.

Von Recklinghausen neurofibromatosis, or type 1 neurofibromatosis (NF1), is a common autosomal dominant disorder characterized by abnormalities in multiple tissues derived from the embryonic neural crest. Portions of the gene have been recently identified by positional cloning, and sequence analysis has shown homology to the GTPase activating protein (GAP) family. In this report we present the results of an extensive cDNA walk resulting in the cloning of the complete coding region of the NF1 transcript. Analysis of the sequences reveals an open reading frame of 2818 amino acids, although alternatively spliced products may code for different protein isoforms. The gene extends for approximately 300 kb on chromosome 17, with its promoter in a CpG-rich island.

Authors
Marchuk, DA; Saulino, AM; Tavakkol, R; Swaroop, M; Wallace, MR; Andersen, LB; Mitchell, AL; Gutmann, DH; Boguski, M; Collins, FS
MLA Citation
Marchuk, DA, Saulino, AM, Tavakkol, R, Swaroop, M, Wallace, MR, Andersen, LB, Mitchell, AL, Gutmann, DH, Boguski, M, and Collins, FS. "cDNA cloning of the type 1 neurofibromatosis gene: complete sequence of the NF1 gene product." Genomics 11.4 (December 1991): 931-940.
PMID
1783401
Source
pubmed
Published In
Genomics
Volume
11
Issue
4
Publish Date
1991
Start Page
931
End Page
940

A highly polymorphic cDNA probe in the NF1 gene.

Authors
Andersen, LB; Wallace, MR; Marchuk, DA; Tavakkol, R; Mitchell, A; Saulino, AM; Collins, FS
MLA Citation
Andersen, LB, Wallace, MR, Marchuk, DA, Tavakkol, R, Mitchell, A, Saulino, AM, and Collins, FS. "A highly polymorphic cDNA probe in the NF1 gene." Nucleic Acids Res 19.13 (July 11, 1991): 3754-.
PMID
1677185
Source
pubmed
Published In
Nucleic Acids Research
Volume
19
Issue
13
Publish Date
1991
Start Page
3754

cDNA sequence and genomic structure of EV12B, a gene lying within an intron of the neurofibromatosis type 1 gene.

The gene responsible for neurofibromatosis type 1 (NF1), one of the more common inherited human disorders, was identified recently, and segments of it were cloned. Two translocation breakpoints that interrupt the NF1 gene in NF1 patients flank a 60-kb segment of DNA that contains the EV12A locus (previously reported as the EV12 locus), the human homolog of a mouse gene, Evi-2A, implicated in retrovirus-induced murine myeloid tumors. EVI2A lies within an intron of the NF1 gene and is transcribed from telomere toward centromere, opposite to the direction of transcription of the NF1 gene. Here we describe a second locus, EVI2B, also located between the two NF1 translocation breakpoints. Full-length cDNAs from the EV12B locus detect a 2.1-kb transcript in bone marrow, peripheral blood mononuclear cells, and fibroblasts. Sequencing studies predict an EV12B protein of 448 amino acids that is proline-rich and contains an N-terminal signal peptide, an extracellular domain with four potential glycosylation sites, a single hydrophobic transmembrane domain, and a cytoplasmic hydrophilic domain. At the level of genomic DNA the EV12B locus lies within the same intron of the NF1 gene as EV12A and contains a 57-bp 5' exon that is noncoding, an 8-kb intron, and a 2078-bp 3' exon that includes the entire open reading frame. EV12B is transcribed in the same direction as EV12A; its 5' exon lies only 4 kb downstream from the 3' exon of the EV12A locus. In the mouse the 5' exon of the homologous gene, Evi-2B, lies approximately 2.8 kb from the 3' end of Evi-2A, in the midst of a cluster of viral integration sites identified in retrovirus-induced myeloid tumors; thus, Evi-2B may function as an oncogene in these tumors.

Authors
Cawthon, RM; Andersen, LB; Buchberg, AM; Xu, GF; O'Connell, P; Viskochil, D; Weiss, RB; Wallace, MR; Marchuk, DA; Culver, M
MLA Citation
Cawthon, RM, Andersen, LB, Buchberg, AM, Xu, GF, O'Connell, P, Viskochil, D, Weiss, RB, Wallace, MR, Marchuk, DA, and Culver, M. "cDNA sequence and genomic structure of EV12B, a gene lying within an intron of the neurofibromatosis type 1 gene." Genomics 9.3 (March 1991): 446-460.
PMID
1903357
Source
pubmed
Published In
Genomics
Volume
9
Issue
3
Publish Date
1991
Start Page
446
End Page
460

A polymorphic cDNA probe on chromosome 17q11.2 located within the NF1 gene [D17S376].

Authors
Andersen, LB; Wallace, MR; Marchuk, DA; Cawthon, RM; Odeh, HM; Letcher, R; White, RL; Collins, FS
MLA Citation
Andersen, LB, Wallace, MR, Marchuk, DA, Cawthon, RM, Odeh, HM, Letcher, R, White, RL, and Collins, FS. "A polymorphic cDNA probe on chromosome 17q11.2 located within the NF1 gene [D17S376]." Nucleic Acids Res 19.1 (January 11, 1991): 197-.
PMID
1672744
Source
pubmed
Published In
Nucleic Acids Research
Volume
19
Issue
1
Publish Date
1991
Start Page
197

Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products

Authors
Marchuk, D; Drumm, M; Saulino, A; Collins, FS
MLA Citation
Marchuk, D, Drumm, M, Saulino, A, and Collins, FS. "Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products." Nucleic Acids Research 19.5 (1991): 1154--.
PMID
2020552
Source
scival
Published In
Nucleic Acids Research
Volume
19
Issue
5
Publish Date
1991
Start Page
1154-

A 90 kb DNA deletion associated with neurofibromatosis type 1.

A deletion of 90 kb of DNA has been identified in a patient with neurofibromatosis type 1, using pulsed field gel electrophoresis. The deletion lies between probes 17L1A and AC5 in the critical region of chromosome 17 and represents the only molecular alteration found by PFGE in a series of 90 unrelated patients. The subject showing the deletion is an isolated case, shows typical clinical features, and represents one of the first examples of a molecular deletion to be found in this disorder.

Authors
Upadhyaya, M; Cheryson, A; Broadhead, W; Fryer, A; Shaw, DJ; Huson, S; Wallace, MR; Andersen, LB; Marchuk, DA; Viskochil, D
MLA Citation
Upadhyaya, M, Cheryson, A, Broadhead, W, Fryer, A, Shaw, DJ, Huson, S, Wallace, MR, Andersen, LB, Marchuk, DA, and Viskochil, D. "A 90 kb DNA deletion associated with neurofibromatosis type 1." J Med Genet 27.12 (December 1990): 738-741.
PMID
2127432
Source
pubmed
Published In
Journal of medical genetics
Volume
27
Issue
12
Publish Date
1990
Start Page
738
End Page
741

A chromosome jump crosses a translocation breakpoint in the von Recklinghausen neurofibromatosis region.

The von Recklinghausen neurofibromatosis (NFI) gene has been previously localized to the region 17q11.2 by genetic analysis. Consistent with this, two NFI patients have been described with autosomal translocations with breakpoints in 17q11.2, and these represent presumed markers for the location of the NFI gene. Recent work has defined the two breakpoints on a physical map, and they lie less than 100 kb apart. To characterize further the distance between these breakpoints and clone additional DNA, a chromosome jump was made from a DNA fragment that maps between the breakpoints. The end of the jump crosses one of the NFI translocation breakpoints and detects that breakpoint on Southern analysis, placing the probe less than 15 kb telomeric to this breakpoint. Pulsed field analysis with the jump clone allows revision of the previous NFI region map and indicates that the two breakpoints lie no more than 60 kb apart. This jump clone will be useful for further mapping, breakpoint cloning, analysis of patient DNA, and the search for transcripts in the NFI region.

Authors
Wallace, MR; Andersen, LB; Fountain, JW; Odeh, HM; Viskochil, D; Marchuk, DA; O'Connell, P; White, R; Collins, FS
MLA Citation
Wallace, MR, Andersen, LB, Fountain, JW, Odeh, HM, Viskochil, D, Marchuk, DA, O'Connell, P, White, R, and Collins, FS. "A chromosome jump crosses a translocation breakpoint in the von Recklinghausen neurofibromatosis region." Genes Chromosomes Cancer 2.4 (November 1990): 271-277.
PMID
2176541
Source
pubmed
Published In
Genes, Chromosomes and Cancer
Volume
2
Issue
4
Publish Date
1990
Start Page
271
End Page
277

Type 1 neurofibromatosis gene: identification of a large transcript disrupted in three NF1 patients.

Von Recklinghausen neurofibromatosis (NF1) is a common autosomal dominant disorder characterized by abnormalities in multiple tissues derived from the neural crest. No reliable cellular phenotypic marker has been identified, which has hampered direct efforts to identify the gene. The chromosome location of the NF1 gene has been previously mapped genetically to 17q11.2, and data from two NF1 patients with balanced translocations in this region have further narrowed the candidate interval. The use of chromosome jumping and yeast artificial chromosome technology has now led to the identification of a large (approximately 13 kilobases) ubiquitously expressed transcript (denoted NF1LT) from this region that is definitely interrupted by one and most likely by both translocations. Previously identified candidate genes, which failed to show abnormalities in NF1 patients, are apparently located within introns of NF1LT, on the antisense strand. A new mutation patient with NF1 has been identified with a de novo 0.5-kilobase insertion in the NF1LT gene. These observations, together with the high spontaneous mutation rate of NF1 (which is consistent with a large locus), suggest that NF1LT represents the elusive NF1 gene.

Authors
Wallace, MR; Marchuk, DA; Andersen, LB; Letcher, R; Odeh, HM; Saulino, AM; Fountain, JW; Brereton, A; Nicholson, J; Mitchell, AL
MLA Citation
Wallace, MR, Marchuk, DA, Andersen, LB, Letcher, R, Odeh, HM, Saulino, AM, Fountain, JW, Brereton, A, Nicholson, J, and Mitchell, AL. "Type 1 neurofibromatosis gene: identification of a large transcript disrupted in three NF1 patients." Science 249.4965 (July 13, 1990): 181-186.
PMID
2134734
Source
pubmed
Published In
Science
Volume
249
Issue
4965
Publish Date
1990
Start Page
181
End Page
186

Type 1 neurofibromatosis gene: Correction (I)

Authors
Wallace, MR; Marchuk, DA; Andersen, LB; Collins, FS
MLA Citation
Wallace, MR, Marchuk, DA, Andersen, LB, and Collins, FS. "Type 1 neurofibromatosis gene: Correction (I)." Science 250.4988 (1990): 1749--.
Source
scival
Published In
Science
Volume
250
Issue
4988
Publish Date
1990
Start Page
1749-

The NF1 locus encodes a protein functionally related to mammalian GAP and yeast IRA proteins

The von Recklinghausen neurofibromatosis locus, NF1, encodes a protein with homology restricted to the catalytic region of the RAS GTPase-activating protein, GAP, and with extensive homology to the IRA1 and IRA2 gene products of the yeast S. cerevisiae. A segment of the NF1 cDNA gene, expressed in yeast, can complement loss of IRA function and can inhibit both wild-type and mutant activated human H-ras genes that are coexpressed in yeast. Yeast expressing the NF1 segment have increased H-ras GTPase-stimulating activity. These studies indicate that the NF1 gene product can interact with RAS proteins and demonstrate structural and functional similarities and differences among the GAP, IRA1, IRA2, and NF1 proteins.

Authors
Ballester, R; Marchuk, D; Boguski, M; Saulino, A; Letcher, R; Wigler, M; Collins, F
MLA Citation
Ballester, R, Marchuk, D, Boguski, M, Saulino, A, Letcher, R, Wigler, M, and Collins, F. "The NF1 locus encodes a protein functionally related to mammalian GAP and yeast IRA proteins." Cell 63.4 (1990): 851-859.
PMID
2121371
Source
scival
Published In
Cell
Volume
63
Issue
4
Publish Date
1990
Start Page
851
End Page
859
DOI
10.1016/0092-8674(90)90151-4

pYAC-RC, a yeast artificial chromosome vector for clonlng DNA cut with infrequently cutting restriction eodonucleases

Authors
Marchuk, D; Collins, FS
MLA Citation
Marchuk, D, and Collins, FS. "pYAC-RC, a yeast artificial chromosome vector for clonlng DNA cut with infrequently cutting restriction eodonucleases." Nucleic Acids Research 16.15 (1988): 7743--.
PMID
3045765
Source
scival
Published In
Nucleic Acids Research
Volume
16
Issue
15
Publish Date
1988
Start Page
7743-
DOI
10.1093/nar/16.15.7743

The human keratin genes and their differential expression.

Authors
Fuchs, E; Tyner, AL; Giudice, GJ; Marchuk, D; RayChaudhury, A; Rosenberg, M
MLA Citation
Fuchs, E, Tyner, AL, Giudice, GJ, Marchuk, D, RayChaudhury, A, and Rosenberg, M. "The human keratin genes and their differential expression." Current topics in developmental biology 22 (1987): 5-34.
PMID
2443316
Source
scival
Published In
Current topics in developmental biology
Volume
22
Publish Date
1987
Start Page
5
End Page
34

Three tightly linked genes encoding human type I keratins: Conservation of sequence in the 5'-untranslated leader and 5'-upstream regions of coexpressed keratin genes

Authors
RayChaudhury, A; Marchuk, D; Lindhurst, M; Fuchs, E
MLA Citation
RayChaudhury, A, Marchuk, D, Lindhurst, M, and Fuchs, E. "Three tightly linked genes encoding human type I keratins: Conservation of sequence in the 5'-untranslated leader and 5'-upstream regions of coexpressed keratin genes." Molecular and Cellular Biology 6.2 (1986): 539-548.
PMID
2431270
Source
scival
Published In
Molecular and Cellular Biology
Volume
6
Issue
2
Publish Date
1986
Start Page
539
End Page
548

Complete sequence of a gene encoding a human type I keratin: Sequences homologous to enhancer elements in the regulatory region of the gene

We report here the complete nucelotide sequence of a gene encoding the 50-kDa keratin expressed in abundance in human epidermal cells. According to its sequence, this gene has a single transcriptional initiation site and a single polyadenylylation signal. Nuclease S1 mapping of this gene with total human epidermal mRNA confirmed the presence of a single initiation site for the 50-kDa keratin gene. When the regulatory sequences 5' upstream from this gene were examined, three sequences that share significant homology with viral and immunoglobulin enhancer elements were found. In comparison, the sequence of the regulatory region of vimentin, a structurally similar intermediate filament gene, was highly divergent [Quax, W., Egberts, W.V., Hendriks, W., Quax-Jeuken, Y. & Bloemendal, H. (1983) Cell 35, 215-223]. This finding may provide a clue to understanding the molecular mechanisms underlying the widely varying levels of expression of different intermediate filament genes in different tissues.

Authors
Marchuk, D; McCrohon, S; Fuchs, E
MLA Citation
Marchuk, D, McCrohon, S, and Fuchs, E. "Complete sequence of a gene encoding a human type I keratin: Sequences homologous to enhancer elements in the regulatory region of the gene." Proceedings of the National Academy of Sciences of the United States of America 82.6 (1985): 1609-1613.
PMID
2580298
Source
scival
Published In
Proceedings of the National Academy of Sciences of the United States of America
Volume
82
Issue
6
Publish Date
1985
Start Page
1609
End Page
1613

The nature and significance of differential keratin gene expression.

Authors
Fuchs, E; Hanukoglu, I; Marchuk, D; Grace, MP; Kim, KH
MLA Citation
Fuchs, E, Hanukoglu, I, Marchuk, D, Grace, MP, and Kim, KH. "The nature and significance of differential keratin gene expression." Annals of the New York Academy of Sciences 455 (1985): 436-450.
PMID
2417525
Source
scival
Published In
Annals of the New York Academy of Sciences
Volume
455
Publish Date
1985
Start Page
436
End Page
450

Expression of unusually large keratins during terminal differentiation: Balance of type I and type II keratins is not disrupted

When a basal epidermal cell undergoes a commitment to terminally differentiate, it ceases to divide and begins to migrate outward towards the surface of the skin. Dramatic changes in its cytoskeletal architecture take place, accompanied by numerous changes in the expression of keratins, a family of related polypeptides that form 8-nm filaments in these cells. We show here that a shift to the synthesis of unusually large keratins occurs that does not seem to disrupt the ratio of two distinct subfamilies of keratins. Preliminary studies indicate that this differentiation-specific shift may be at the level of transcriptional rather than post-transcriptional regulation. The striking similarities between these large keratins and the type I and type II keratins of basal epidermal cells suggests the important role that both classes of large keratin sequences must play in the assembly of the intermediate filaments within the differentiating keratinocyte.

Authors
Kim, KH; Marchuk, D; Fuchs, E
MLA Citation
Kim, KH, Marchuk, D, and Fuchs, E. "Expression of unusually large keratins during terminal differentiation: Balance of type I and type II keratins is not disrupted." Journal of Cell Biology 99.5 (1984): 1872-1877.
PMID
6208205
Source
scival
Published In
Journal of Cell Biology
Volume
99
Issue
5
Publish Date
1984
Start Page
1872
End Page
1877

Remarkable conservation of structure among intermediate filament genes

Using a cloned cDNA complementary to a portion of the mRNA for the 50 kd human epidermal keratin, we have screened a human genomic library and have isolated and sequenced the gene encoding this keratin. A comparison of the keratin gene with the very distantly related vimentin gene has enabled us to explore the relation between the evolutionary conservation of structure in intermediate filament (IF) subunits and the conservation of structure in IF genes. Our results reveal that not only the secondary structure of the IF proteins, but also the structural skeleton of their genes, has been maintained throughout evolution. These characteristics have persisted despite considerable flexibility in both protein and nucleic acid sequence. Surprisingly, although the positions of the introns within these two genes are highly conserved, they do not seem to correspond to the boundaries of the structural domains common to all IF subunits. © 1984.

Authors
Marchuk, D; McCrohon, S; Fuchs, E
MLA Citation
Marchuk, D, McCrohon, S, and Fuchs, E. "Remarkable conservation of structure among intermediate filament genes." Cell 39.3 PART 2 (1984): 491-498.
PMID
6210150
Source
scival
Published In
Cell
Volume
39
Issue
3 PART 2
Publish Date
1984
Start Page
491
End Page
498
DOI
10.1016/0092-8674(84)90456-2

Type I and type II keratins have evolved from lower eukaryotes to form the epidermal intermediate filaments in mammalian skin

We have traced the evolutionary origins of keratin-like sequences to the genomes of lower eukaryotes. The proteins encoded by these genes have evolved from the intermediate filaments that comprise the backbone of vertebrate skin cells. Two related but distinct types of keratins encoded by two separate multigene subfamilies are expressed in the epidermal keratinocytes of vertebrate species from fish to human. Both at the level of protein and at the level of DNA, these two classes of keratins are coordinately conserved throughout vertebrate evolution, indicating the central role that both types of keratins must play in the assembly and structure of the 8-nm filament.

Authors
Fuchs, E; Marchuk, D
MLA Citation
Fuchs, E, and Marchuk, D. "Type I and type II keratins have evolved from lower eukaryotes to form the epidermal intermediate filaments in mammalian skin." Proceedings of the National Academy of Sciences of the United States of America 80.19 I (1983): 5857-5861.
PMID
6193525
Source
scival
Published In
Proceedings of the National Academy of Sciences of the United States of America
Volume
80
Issue
19 I
Publish Date
1983
Start Page
5857
End Page
5861
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