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Datto, Michael Bradley

Overview:

Dr. Datto is an AP/CP/MGP board certified pathologist who specializes in molecular pathology. He is the Medical Director for Duke University Health System Clinical Laboratories.  He also directs the Duke University Health System Clinical Molecular Diagnostics Laboratory. In these roles, he is responsible for maintaining the standards of the College of American Pathologists and CLIA/CMS within all Clinical Laboratories at Duke.  Specifically, Dr. Datto oversees clinical testing and reporting, develops quality management systems and proficiency testing programs, provides consultation with ordering physicians, ensures educational programs, develops strategic plans that are in line with the needs of our patient population, physicians and health system leadership, coordinates research and development, ensures adequate and appropriately trained personnel, and provides profession interpretation for molecular diagnostic testing including the wide range of PCR, quantitative PCR, sequencing and FISH based tests for inherited genetic diseases, hematologic malignancies, solid tumors and infectious diseases.



Dr. Datto also has an independent research program developing and evaluating multigene assays to determine cancer patient specific prognoses in the context of breast, ovary and colon cancer. The ultimate goal of this work is to produce improved molecular diagnostic assays that incorporate genome wide prognostic and diagnostic molecular profiling into the clinical laboratories for routine patient care. This involves the study of factors that affect the precision and accuracy of array-based and next-generation sequencing based approaches, as well as the discovery of novel diagnostic molecular makers. Finally, Dr. Datto has a strong research interest in resident and medical student education, specifically the development of web-base community tools for anatomic and clinical pathology education.

Positions:

Associate Professor of Pathology

Pathology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

B.A. 1991

B.A. — Johns Hopkins University

Ph.D. 1998

Ph.D. — Duke University

M.D. 1999

M.D. — Duke University

Post Doctoral Fellow, Pharmacology

Duke University

Resident, Pathology

Duke University

Grants:

Non Muscle Myosin II Contractility Putatively Regulates Scar Contracture

Administered By
Surgery
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
September 15, 2008
End Date
August 31, 2013

Programs in Clinical Effectiveness of Cancer Pharmacogenomics

Administered By
Duke Center for Applied Genomics and Precision Medicine
AwardedBy
National Institutes of Health
Role
Pathologist
Start Date
September 30, 2009
End Date
August 31, 2012

Validating Molecular Signature Risk Models of NSCLC

Administered By
Surgery, Cardiovascular and Thoracic Surgery
AwardedBy
National Institutes of Health
Role
Pathologist
Start Date
August 21, 2006
End Date
July 31, 2012

Intellectual Property Challenges for the Development of Genomic Diagnostics

Administered By
University Institutes and Centers
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
April 01, 2009
End Date
March 31, 2012

Gene Targeted Therapy of Brain Tumors

Administered By
Neurosurgery
AwardedBy
National Institutes of Health
Role
Associate Research Professor
Start Date
September 30, 2009
End Date
February 29, 2012

Prospective Validation of Genomic Signatures of Chemosensitivity in NSCLC

Administered By
Institutes and Centers
AwardedBy
National Institutes of Health
Role
Investigator
Start Date
January 01, 2009
End Date
November 30, 2010
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Publications:

Reply to Young Investigator Challenge: Molecular testing in noninvasive follicular thyroid neoplasm with papillary-like nuclear features.

Authors
Jiang, XS; Harrison, GP; Datto, MB
MLA Citation
Jiang, XS, Harrison, GP, and Datto, MB. "Reply to Young Investigator Challenge: Molecular testing in noninvasive follicular thyroid neoplasm with papillary-like nuclear features." Cancer 125.4 (April 2017): 293-294. (Letter)
PMID
28170161
Source
epmc
Published In
Cancer
Volume
125
Issue
4
Publish Date
2017
Start Page
293
End Page
294
DOI
10.1002/cncy.21829

Standards and Guidelines for the Interpretation and Reporting of Sequence Variants in Cancer: A Joint Consensus Recommendation of the Association for Molecular Pathology, American Society of Clinical Oncology, and College of American Pathologists.

Widespread clinical laboratory implementation of next-generation sequencing-based cancer testing has highlighted the importance and potential benefits of standardizing the interpretation and reporting of molecular results among laboratories. A multidisciplinary working group tasked to assess the current status of next-generation sequencing-based cancer testing and establish standardized consensus classification, annotation, interpretation, and reporting conventions for somatic sequence variants was convened by the Association for Molecular Pathology with liaison representation from the American College of Medical Genetics and Genomics, American Society of Clinical Oncology, and College of American Pathologists. On the basis of the results of professional surveys, literature review, and the Working Group's subject matter expert consensus, a four-tiered system to categorize somatic sequence variations based on their clinical significances is proposed: tier I, variants with strong clinical significance; tier II, variants with potential clinical significance; tier III, variants of unknown clinical significance; and tier IV, variants deemed benign or likely benign. Cancer genomics is a rapidly evolving field; therefore, the clinical significance of any variant in therapy, diagnosis, or prognosis should be reevaluated on an ongoing basis. Reporting of genomic variants should follow standard nomenclature, with testing method and limitations clearly described. Clinical recommendations should be concise and correlate with histological and clinical findings.

Authors
Li, MM; Datto, M; Duncavage, EJ; Kulkarni, S; Lindeman, NI; Roy, S; Tsimberidou, AM; Vnencak-Jones, CL; Wolff, DJ; Younes, A; Nikiforova, MN
MLA Citation
Li, MM, Datto, M, Duncavage, EJ, Kulkarni, S, Lindeman, NI, Roy, S, Tsimberidou, AM, Vnencak-Jones, CL, Wolff, DJ, Younes, A, and Nikiforova, MN. "Standards and Guidelines for the Interpretation and Reporting of Sequence Variants in Cancer: A Joint Consensus Recommendation of the Association for Molecular Pathology, American Society of Clinical Oncology, and College of American Pathologists." The Journal of molecular diagnostics : JMD 19.1 (January 2017): 4-23. (Review)
PMID
27993330
Source
epmc
Published In
The Journal of molecular diagnostics : JMD
Volume
19
Issue
1
Publish Date
2017
Start Page
4
End Page
23
DOI
10.1016/j.jmoldx.2016.10.002

Young Investigator Challenge: Molecular testing in noninvasive follicular thyroid neoplasm with papillary-like nuclear features.

Molecular testing provides an important ancillary study for thyroid nodules with indeterminate cytology. The nomenclature shift to "noninvasive follicular thyroid neoplasm with papillary-like nuclear features" (NIFTP) will impact the performance of molecular tests. For the current study, the authors reviewed the performance of the Afirma gene-expression classifier (GEC) and the University of Pittsburgh Medical Center (UPMC) targeted mutation panel tests in thyroid nodules that were subsequently diagnosed as NIFTP on surgical resection.In total, 302 nodules were sent for molecular testing between June 2012 and June 2016. These cases were retrospectively reviewed to identify patients who underwent subsequent surgical resection and were diagnosed with follicular variant of papillary thyroid carcinoma (FVPTC). Twenty-five nodules that were diagnosed as FVPTC met the initial inclusion criteria. These cases were reviewed using strict criteria to identify NIFTP.Eight cases met criteria for NIFTP, and 4 NIFTPs underwent Afirma testing. Cytology diagnoses were all Bethesda category III, with 3 diagnosed as atypia of undetermined significance (AUS) and 1 diagnosed as follicular lesion of undetermined significance (FLUS). All of these nodules were identified as "suspicious" using GEC. Four NIFTPs underwent testing at UPMC, all using ThyroSeq V2. The cytology diagnoses for these nodules also were category III, with the exception of 1 nodule that was category IV, suspicious for follicular neoplasm. All NIFTPs were positive for mutations, all of which were RAS mutations (NRAS, KRAS). One patient who had a nodule classified as NIFTP had metastatic carcinoma identified in a lymph node. Another who had a 6-cm tumor had coexisting NRAS and TERT mutations.The current results indicate that NIFTP is a rare tumor if defined by strict criteria, that both the GEC and UPMC methods indicate abnormalities in NIFTP, and further independent study will be needed to better characterize the molecular and clinical characteristics of NIFTP. Cancer Cytopathol 2016;124:893-900. © 2016 American Cancer Society.

Authors
Jiang, XS; Harrison, GP; Datto, MB
MLA Citation
Jiang, XS, Harrison, GP, and Datto, MB. "Young Investigator Challenge: Molecular testing in noninvasive follicular thyroid neoplasm with papillary-like nuclear features." Cancer 124.12 (December 2016): 893-900.
PMID
27893191
Source
epmc
Published In
Cancer
Volume
124
Issue
12
Publish Date
2016
Start Page
893
End Page
900
DOI
10.1002/cncy.21802

Molecular testing for the BRCA1 and BRCA2 Ashkenazi Jewish founder mutations: a report on the College of American Pathologists proficiency testing surveys.

The purpose of this study was to analyze laboratory performance on proficiency testing surveys offered jointly by the College of American Pathologists/American College of Medical Genetics and Genomics biannually for the three common Ashkenazi Jewish founder mutations in the BRCA1 and BRCA2 genes.Survey responses were analyzed for accuracy of genotype determination and the associated clinical interpretation. Data on an individual laboratory's participation over time, number of samples tested, turnaround time, and test methodology were also reviewed.Between 2003 and 2012, 23 US laboratories and 39 international laboratories participated. There were six genotyping errors, with a corresponding analytical sensitivity of 99.0% (479/484 challenges; 95% confidence interval: 97.6-99.7%) and an analytic specificity of 99.9% (870/871; 95% confidence interval: 99.4-99.9%). Among the 1,325 clinical interpretations, 92.5% (1,226/1,325; 95% confidence interval: 91.0-93.9%) matched the intended response. Most of the 99 discrepancies-81% (80/99)-incorrectly interpreted the risk for a negative test result as having a lifetime risk of breast cancer "that is the same as that in the general population" instead of "that cannot be determined without BRCA mutation testing of the affected relative."Clinical laboratories demonstrated excellent analytical sensitivity and specificity. The clinical interpretation requires additional education, focusing on the clinical interpretation of negative test results for these three mutations.

Authors
Tafe, LJ; Datto, MB; Palomaki, GE; Lacbawan, FL; CAP/ACMG Biochemical and Molecular Genetics Resource Committee,
MLA Citation
Tafe, LJ, Datto, MB, Palomaki, GE, Lacbawan, FL, and CAP/ACMG Biochemical and Molecular Genetics Resource Committee, . "Molecular testing for the BRCA1 and BRCA2 Ashkenazi Jewish founder mutations: a report on the College of American Pathologists proficiency testing surveys." Genetics in medicine : official journal of the American College of Medical Genetics 17.1 (January 2015): 58-62.
PMID
24946157
Source
epmc
Published In
Genetics in Medicine
Volume
17
Issue
1
Publish Date
2015
Start Page
58
End Page
62
DOI
10.1038/gim.2014.77

False positives in multiplex PCR-based next-generation sequencing have unique signatures.

Next-generation sequencing shows great promise by allowing rapid mutational analysis of multiple genes in human cancers. Recently, we implemented the multiplex PCR-based Ion AmpliSeq Cancer Hotspot Panel (>200 amplicons in 50 genes) to evaluate EGFR, KRAS, and BRAF in lung and colorectal adenocarcinomas. In 10% of samples, automated analysis identified a novel G873R substitution mutation in EGFR. By examining reads individually, we found this mutation in >5% of reads in 50 of 291 samples and also found similar events in 18 additional amplicons. These apparent mutations are present only in short reads and within 10 bases of either end of the read. We therefore hypothesized that these were from panel primers promiscuously binding to nearly complementary sequences of nontargeted amplicons. Sequences around the mutations matched primer binding sites in the panel in 18 of 19 cases, thus likely corresponding to panel primers. Furthermore, because most primers did not show this effect, we demonstrated that next-generation sequencing may be used to better design multiplex PCR primers through iterative elimination of offending primers to minimize mispriming. Our results indicate the need for careful sequence analysis to avoid false-positive mutations that can arise in multiplex PCR panels. The AmpliSeq Cancer panel is a valuable tool for clinical diagnostics, provided awareness of potential artifacts.

Authors
McCall, CM; Mosier, S; Thiess, M; Debeljak, M; Pallavajjala, A; Beierl, K; Deak, KL; Datto, MB; Gocke, CD; Lin, M-T; Eshleman, JR
MLA Citation
McCall, CM, Mosier, S, Thiess, M, Debeljak, M, Pallavajjala, A, Beierl, K, Deak, KL, Datto, MB, Gocke, CD, Lin, M-T, and Eshleman, JR. "False positives in multiplex PCR-based next-generation sequencing have unique signatures." The Journal of molecular diagnostics : JMD 16.5 (September 2014): 541-549.
PMID
25017478
Source
epmc
Published In
The Journal of molecular diagnostics : JMD
Volume
16
Issue
5
Publish Date
2014
Start Page
541
End Page
549
DOI
10.1016/j.jmoldx.2014.06.001

Tumor acquisition for biomarker research in lung cancer.

The biopsy collection data from two lung cancer trials that required fresh tumor samples be obtained for microarray analysis were reviewed. In the trial for advanced disease, microarray data were obtained on 50 patient samples, giving an overall success rate of 60.2%. The majority of the specimens were obtained through CT-guided lung biopsies (N = 30). In the trial for early-stage patients, 28 tissue specimens were collected from excess tumor after surgical resection with a success rate of 85.7%. This tissue procurement program documents the feasibility in obtaining fresh tumor specimens prospectively that could be used for molecular testing.

Authors
Stevenson, M; Christensen, J; Shoemaker, D; Foster, T; Barry, WT; Tong, BC; Wahidi, M; Shofer, S; Datto, M; Ginsburg, G; Crawford, J; D'Amico, T; Ready, N
MLA Citation
Stevenson, M, Christensen, J, Shoemaker, D, Foster, T, Barry, WT, Tong, BC, Wahidi, M, Shofer, S, Datto, M, Ginsburg, G, Crawford, J, D'Amico, T, and Ready, N. "Tumor acquisition for biomarker research in lung cancer." Cancer investigation 32.6 (July 2014): 291-298.
PMID
24810245
Source
epmc
Published In
Cancer Investigation (Informa)
Volume
32
Issue
6
Publish Date
2014
Start Page
291
End Page
298
DOI
10.3109/07357907.2014.911880

Developing patient-friendly genetic and genomic test reports: formats to promote patient engagement and understanding.

With the emergence of electronic medical records and patient portals, patients are increasingly able to access their health records, including laboratory reports. However, laboratory reports are usually written for clinicians rather than patients, who may not understand much of the information in the report. While several professional guidelines define the content of test reports, there are no guidelines to inform the development of a patient-friendly laboratory report. In this Opinion, we consider patient barriers to comprehension of lab results and suggest several options to reformat the lab report to promote understanding of test results and their significance to patient care, and to reduce patient anxiety and confusion. In particular, patients' health literacy, genetic literacy, e-health literacy and risk perception may influence their overall understanding of lab results and affect patient care. We propose four options to reformat lab reports: 1) inclusion of an interpretive summary section, 2) a summary letter to accompany the lab report, 3) development of a patient user guide to be provided with the report, and 4) a completely revised patient-friendly report. The complexity of genetic and genomic test reports poses a major challenge to patient understanding that warrants the development of a report more appropriate for patients.

Authors
Haga, SB; Mills, R; Pollak, KI; Rehder, C; Buchanan, AH; Lipkus, IM; Crow, JH; Datto, M
MLA Citation
Haga, SB, Mills, R, Pollak, KI, Rehder, C, Buchanan, AH, Lipkus, IM, Crow, JH, and Datto, M. "Developing patient-friendly genetic and genomic test reports: formats to promote patient engagement and understanding." Genome medicine 6.7 (January 2014): 58-.
PMID
25473429
Source
epmc
Published In
Genome Medicine: medicine in the post-genomic era
Volume
6
Issue
7
Publish Date
2014
Start Page
58
DOI
10.1186/s13073-014-0058-6

Report of a young girl with MYH9 mutation and review of the literature.

MYH9 mutations cause the inherited macro-thrombocytopenic syndromes of May-Hegglin anomaly, Fechtner syndrome, Sebastian syndrome, and Epstein syndrome, collectively referred to as MYH9-related disease. We present the case of a girl with MYH9-related disease whose diagnosis was facilitated by platelet electron microscopy and MYH9 sequencing. We discuss our patient's clinical presentation, now with 12 years of follow-up. We also discuss management and her possible prognosis given her specific MYH9 mutation.

Authors
Landi, D; Lockhart, E; Miller, SE; Datto, M; Rehder, C; Kanaly, A; Thornburg, CD
MLA Citation
Landi, D, Lockhart, E, Miller, SE, Datto, M, Rehder, C, Kanaly, A, and Thornburg, CD. "Report of a young girl with MYH9 mutation and review of the literature." J Pediatr Hematol Oncol 34.7 (October 2012): 538-540. (Review)
PMID
23007341
Source
pubmed
Published In
Journal of Pediatric Hematology/Oncology
Volume
34
Issue
7
Publish Date
2012
Start Page
538
End Page
540
DOI
10.1097/MPH.0b013e3182678fc9

Characterization of an oxaliplatin sensitivity predictor in a preclinical murine model of colorectal cancer.

Despite advances in contemporary chemotherapeutic strategies, long-term survival still remains elusive for patients with metastatic colorectal cancer. A better understanding of the molecular markers of drug sensitivity to match therapy with patient is needed to improve clinical outcomes. In this study, we used in vitro drug sensitivity data from the NCI-60 cell lines together with their Affymetrix microarray data to develop a gene expression signature to predict sensitivity to oxaliplatin. To validate our oxaliplatin sensitivity signature, patient-derived colorectal cancer explants (PDCCE) were developed in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice from resected human colorectal tumors. Analysis of gene expression profiles found similarities between the PDCCEs and their parental human tumors, suggesting their utility to study drug sensitivity in vivo. The oxaliplatin sensitivity signature was then validated in vivo with response data from 14 PDCCEs treated with oxaliplatin and was found to have an accuracy of 92.9% (sensitivity = 87.5%; specificity = 100%). Our findings suggest that PDCCEs can be a novel source to study drug sensitivity in colorectal cancer. Furthermore, genomic-based analysis has the potential to be incorporated into future strategies to optimize individual therapy for patients with metastatic colorectal cancer.

Authors
Kim, MK; Osada, T; Barry, WT; Yang, XY; Freedman, JA; Tsamis, KA; Datto, M; Clary, BM; Clay, T; Morse, MA; Febbo, PG; Lyerly, HK; Hsu, DS
MLA Citation
Kim, MK, Osada, T, Barry, WT, Yang, XY, Freedman, JA, Tsamis, KA, Datto, M, Clary, BM, Clay, T, Morse, MA, Febbo, PG, Lyerly, HK, and Hsu, DS. "Characterization of an oxaliplatin sensitivity predictor in a preclinical murine model of colorectal cancer." Mol Cancer Ther 11.7 (July 2012): 1500-1509.
PMID
22351745
Source
pubmed
Published In
Molecular cancer therapeutics
Volume
11
Issue
7
Publish Date
2012
Start Page
1500
End Page
1509
DOI
10.1158/1535-7163.MCT-11-0937

Genes with bimodal expression are robust diagnostic targets that define distinct subtypes of epithelial ovarian cancer with different overall survival.

In some cancer types, certain genes behave as molecular switches, with on and off expression states. These genes tend to define tumor subtypes associated with different treatments and different patient survival. We hypothesized that clinically relevant molecular switch genes exist in epithelial ovarian cancer. To test this hypothesis, we applied a bimodal discovery algorithm to a publicly available ovarian cancer expression microarray data set, GSE9891 [285 tumors: 246 malignant serous (MS), 20 endometrioid (EM), and 18 low malignant potential (LMP) ovarian carcinomas]. Genes with robust bimodal expression patterns were identified across all ovarian tumor types and also within selected subtypes: 73 bimodal genes demonstrated differential expression between LMP versus MS and EM; 22 bimodal genes distinguished MS from EM; and 14 genes had significant association with survival among MS tumors. When these genes were combined into a single survival score, the median survival for patients with a favorable versus unfavorable score was 65 versus 29 months (P < 0.0001, hazard ratio = 0.4221). Two independent data sets [high-grade, advanced-stage serous (n = 53) and advanced-stage (n = 119) ovarian tumors] validated the survival score performance. We conclude that genes with bimodal expression patterns not only define clinically relevant molecular subtypes of ovarian carcinoma but also provide ideal targets for translation into the clinical laboratory.

Authors
Kernagis, DN; Hall, AHS; Datto, MB
MLA Citation
Kernagis, DN, Hall, AHS, and Datto, MB. "Genes with bimodal expression are robust diagnostic targets that define distinct subtypes of epithelial ovarian cancer with different overall survival." J Mol Diagn 14.3 (May 2012): 214-222.
PMID
22490445
Source
pubmed
Published In
The Journal of molecular diagnostics : JMD
Volume
14
Issue
3
Publish Date
2012
Start Page
214
End Page
222
DOI
10.1016/j.jmoldx.2012.01.007

Molecular detection of circulating Sezary cells in patients with mycosis fungoides: could it predict future development of secondary Sezary syndrome? A single-institution experience.

While the majority of patients with early-stage mycosis fungoides (MF) have an excellent prognosis, a few cases progress to secondary Sezary syndrome (sSS), which carries a dismal clinical outcome. We retrospectively analyzed 135 cases of MF/SS and correlated molecular detection of T-cell clones in the skin and blood with other clinicopathologic findings. When stratified by the diagnoses, patients with MF demonstrated a 26.5% (31/117) positive rate for a blood T-cell clone, of which 50% (10/20) had an identical T-cell clone in the skin. Follow-up evaluation showed conversion into sSS or leukemic phase in 50% (5/10) of cases with a positive blood T-cell clone (estimated mean interval 41.8 months) in comparison to no cases in the group without a clone (0/31). Interestingly, 4/5 cases of sSS had an identical T-cell clone in the skin, while the remaining case did not have the test performed on skin for clonal comparison. Kaplan-Meier survival analysis demonstrated a poor clinical outcome in the group with a blood T-cell clone, in comparison with the group without, in overall survival (p < 0.0001) and progression-free survival (p < 0.0001; HR = 22.6). These findings suggest that molecular detection of a blood T-cell clone may have a role in predicting sSS. Due to amplification of non-neoplastic T-cell expansion in a significant number of cases, comparison of blood T-cell clones with skin may have confirmatory value.

Authors
Hutchinson, CB; Stoecker, M; Wang, FF; Papalas, J; Sebastian, S; Burchette, J; Datto, M; Wang, E
MLA Citation
Hutchinson, CB, Stoecker, M, Wang, FF, Papalas, J, Sebastian, S, Burchette, J, Datto, M, and Wang, E. "Molecular detection of circulating Sezary cells in patients with mycosis fungoides: could it predict future development of secondary Sezary syndrome? A single-institution experience." Leuk Lymphoma 53.5 (May 2012): 868-877.
PMID
22044137
Source
pubmed
Published In
Leukemia & Lymphoma (Informa)
Volume
53
Issue
5
Publish Date
2012
Start Page
868
End Page
877
DOI
10.3109/10428194.2011.633408

Phase II study of cenersen, an antisense inhibitor of p53, in combination with fludarabine, cyclophosphamide and rituximab for high-risk chronic lymphocytic leukemia.

Patients with chronic lymphocytic leukemia (CLL) with deletion or mutation of TP53 have exceedingly poor clinical outcomes. Cenersen, an oligonucleotide targeting TP53, has been shown to abrogate the activity of TP53 gain-of-function mutants and to increase sensitivity of lymphoma cells to cytotoxic chemotherapy in vitro. We combined cenersen with fludarabine, cyclophosphamide and rituximab (FCR) as treatment for patients with high-risk CLL. The purpose of this phase II study was to determine the overall response rate, response duration and toxicity of cenersen administered in combination with FCR. Twenty patients with relapsed or high-risk CLL were evaluated. Nineteen patients were previously treated. The complete response rate was 18%; the overall response rate was 53%. Median progression-free and overall survival was 5.3 and 10.6 months, respectively. The most common serious adverse events were neutropenia and thrombocytopenia. In this single arm phase II study, cenersen combined with FCR yielded clinical responses with acceptable toxicity in patients with high-risk CLL.

Authors
Lanasa, MC; Davis, PH; Datto, M; Li, Z; Gockerman, JP; Moore, JO; DeCastro, CM; Friedman, DR; Diehl, LF; Rehder, C; Cook, H; Daugherty, FJ; Matta, KMB; Weinberg, JB; Rizzieri, D
MLA Citation
Lanasa, MC, Davis, PH, Datto, M, Li, Z, Gockerman, JP, Moore, JO, DeCastro, CM, Friedman, DR, Diehl, LF, Rehder, C, Cook, H, Daugherty, FJ, Matta, KMB, Weinberg, JB, and Rizzieri, D. "Phase II study of cenersen, an antisense inhibitor of p53, in combination with fludarabine, cyclophosphamide and rituximab for high-risk chronic lymphocytic leukemia." Leuk Lymphoma 53.2 (February 2012): 218-224.
PMID
21827374
Source
pubmed
Published In
Leukemia & Lymphoma (Informa)
Volume
53
Issue
2
Publish Date
2012
Start Page
218
End Page
224
DOI
10.3109/10428194.2011.610012

Molecular genetic testing for fragile X syndrome: Laboratory performance on the College of American Pathologists proficiency surveys (2001-2009)

Purpose: The College of American Pathologists offers biannual proficiency testing for molecular analysis of fragile X syndrome. The purpose of this study was to analyze laboratory performance on the fragile X proficiency surveys from 2001 to 2009. Methods: Individual laboratory responses were analyzed for accuracy of genotype determination (normal, gray zone, premutation, or full mutation) and size analysis of the FMR1 trinucleotide repeat region. The analytical sensitivity and specificity of testing for fragile X were calculated, and laboratory performance for trinucleotide repeat sizing was evaluated. Results: Overall, laboratories demonstrated analytical sensitivity of 99% and 96% for detection of full mutations associated with fragile X syndrome in males and females, respectively; analytical sensitivity of 98% for detection of premutations; and analytical specificity of 99.9%. Size measurements of the CGG repeat region were acceptable from most laboratories, with an increase in the range of reported sizes observed for larger repeat expansions. Conclusions: Molecular genetic testing for fragile X syndrome demonstrated excellent sensitivity and specificity by laboratories participating in the College of American Pathologists (CAP) surveys. Allele sizing demonstrated good performance overall with improved accuracy over the study period. Participation in proficiency testing can aid laboratories in assessing individual performance and need for calibration of assays. © American College of Medical Genetics and Genomics.

Authors
Weck, KE; Zehnbauer, B; Datto, M; Schrijver, I
MLA Citation
Weck, KE, Zehnbauer, B, Datto, M, and Schrijver, I. "Molecular genetic testing for fragile X syndrome: Laboratory performance on the College of American Pathologists proficiency surveys (2001-2009)." Genetics in Medicine 14.3 (2012): 306-312.
PMID
22241100
Source
scival
Published In
Genetics in Medicine
Volume
14
Issue
3
Publish Date
2012
Start Page
306
End Page
312
DOI
10.1038/gim.2011.11

Type III TGF-β receptor enhances colon cancer cell migration and anchorage-independent growth.

The type III TGF-β receptor (TβRIII or betagylcan) is a TGF-β superfamily coreceptor with emerging roles in regulating TGF-β superfamily signaling and cancer progression. Alterations in TGF-β superfamily signaling are common in colon cancer; however, the role of TβRIII has not been examined. Although TβRIII expression is frequently lost at the message and protein level in human cancers and suppresses cancer progression in these contexts, here we demonstrate that, in colon cancer, TβRIII messenger RNA expression is not significantly altered and TβRIII expression is more frequently increased at the protein level, suggesting a distinct role for TβRIII in colon cancer. Increasing TβRIII expression in colon cancer model systems enhanced ligand-mediated phosphorylation of p38 and the Smad proteins, while switching TGF-β and BMP-2 from inhibitors to stimulators of colon cancer cell proliferation, inhibiting ligand-induced p21 and p27 expression. In addition, increasing TβRIII expression increased ligand-stimulated anchorage-independent growth, a resistance to ligand- and chemotherapy-induced apoptosis, cell migration and modestly increased tumorigenicity in vivo. In a reciprocal manner, silencing endogenous TβRIII expression decreased colon cancer cell migration. These data support a model whereby TβRIII mediates TGF-β superfamily ligand-induced colon cancer progression and support a context-dependent role for TβRIII in regulating cancer progression.

Authors
Gatza, CE; Holtzhausen, A; Kirkbride, KC; Morton, A; Gatza, ML; Datto, MB; Blobe, GC
MLA Citation
Gatza, CE, Holtzhausen, A, Kirkbride, KC, Morton, A, Gatza, ML, Datto, MB, and Blobe, GC. "Type III TGF-β receptor enhances colon cancer cell migration and anchorage-independent growth." Neoplasia 13.8 (August 2011): 758-770.
PMID
21847367
Source
pubmed
Published In
Neoplasia (New York, N.Y.)
Volume
13
Issue
8
Publish Date
2011
Start Page
758
End Page
770

Donor cell-derived leukemias/myelodysplastic neoplasms in allogeneic hematopoietic stem cell transplant recipients: a clinicopathologic study of 10 cases and a comprehensive review of the literature.

We report 10 cases of donor cell leukemia (DCL). All cases except the case of chronic lymphocytic leukemia had anemia, neutropenia, and/or thrombocytopenia when DCL was diagnosed. Eight cases with sex-mismatched hematopoietic stem cell transplant (HCT) showed donor gonosomal complements, suggesting DCL. Clonal cytogenetic abnormalities were detected in 8 cases: 6 were monosomy 7/del(7q). In all 10 cases, engraftment studies confirmed donor cell origin. Retrospective fluorescence in situ hybridization in archived donor cells in 4 cases showed a low level of abnormalities in 2. Of 7 patients with clinical follow-up of 5 months or more, 1 (with acute myeloid leukemia) died of disease; 6 are alive, including 1 with myelodysplastic syndrome with spontaneous remission. Similar to reported cases, we found disproportional sex-mismatched HCTs, suggesting probable underdetection of DCL in sex-matched HCTs. The latency between HCT and DCL ranged from 1 to 193 months (median, 24 months), in keeping with the literature. Analyzing our cases, pooled with reported cases, with survival models showed much shorter latency for malignancy as primary disease, for T-cell large granular lymphocyte leukemia as type of DCL, and for umbilical cord blood as stem cell source.

Authors
Wang, E; Hutchinson, CB; Huang, Q; Lu, CM; Crow, J; Wang, FF; Sebastian, S; Rehder, C; Lagoo, A; Horwitz, M; Rizzieri, D; Yu, J; Goodman, B; Datto, M; Buckley, P
MLA Citation
Wang, E, Hutchinson, CB, Huang, Q, Lu, CM, Crow, J, Wang, FF, Sebastian, S, Rehder, C, Lagoo, A, Horwitz, M, Rizzieri, D, Yu, J, Goodman, B, Datto, M, and Buckley, P. "Donor cell-derived leukemias/myelodysplastic neoplasms in allogeneic hematopoietic stem cell transplant recipients: a clinicopathologic study of 10 cases and a comprehensive review of the literature." Am J Clin Pathol 135.4 (April 2011): 525-540. (Review)
PMID
21411775
Source
pubmed
Published In
American Journal of Clinical Pathology
Volume
135
Issue
4
Publish Date
2011
Start Page
525
End Page
540
DOI
10.1309/AJCPPJUQ9DNR1GHP

Sequential development of histiocytic sarcoma and diffuse large b-cell lymphoma in a patient with a remote history of follicular lymphoma with genotypic evidence of a clonal relationship: a divergent (bilineal) neoplastic transformation of an indolent B-cell lymphoma in a single individual.

Follicular lymphoma (FL) often transforms to diffuse large B-cell lymphoma (DLBCL) during its protracted clinical course. Rarely, histiocytic sarcoma (HS) occurs secondary to or concurrent with FL, although the biological relationship between these 2 morphologically and immunophenotypically distinct entities in the same individual has not been well characterized. We report a unique case showing the sequential occurrence of first, HS and then DLBCL in a patient with a remote history of FL. In this case, HS developed 17 years after the diagnosis of FL and was shown to partly retain the immunophenotypic features characteristic of FL, while showing the morphologic and immunophenotypic profiles diagnostic of HS. DLBCL occurred 18.5 years after FL. Both HS and DLBCL harbored the IGH/BCL2 fusion gene, a hallmark of FL, per interphase fluorescence in situ hybridization analysis. Immunoglobulin gene rearrangement studies showed a clonal rearrangement of the IGH gene in both HS and DLBCL with identical amplicons, suggesting a shared origin of the neoplastic clones. These data support the hypothesis of transdifferentiation or transevolution in a mature B-cell neoplasm, and, in addition, suggest the possibility of a divergent (bilineal) neoplastic transformation of FL in a single individual.

Authors
Wang, E; Papalas, J; Hutchinson, CB; Kulbacki, E; Huang, Q; Sebastian, S; Rehder, C; Silbermins, D; Moore, J; Datto, M
MLA Citation
Wang, E, Papalas, J, Hutchinson, CB, Kulbacki, E, Huang, Q, Sebastian, S, Rehder, C, Silbermins, D, Moore, J, and Datto, M. "Sequential development of histiocytic sarcoma and diffuse large b-cell lymphoma in a patient with a remote history of follicular lymphoma with genotypic evidence of a clonal relationship: a divergent (bilineal) neoplastic transformation of an indolent B-cell lymphoma in a single individual." Am J Surg Pathol 35.3 (March 2011): 457-463.
PMID
21317718
Source
pubmed
Published In
American Journal of Surgical Pathology
Volume
35
Issue
3
Publish Date
2011
Start Page
457
End Page
463
DOI
10.1097/PAS.0b013e3182098799

Donor cell leukemia in umbilical cord blood transplant patients: a case study and literature review highlighting the importance of molecular engraftment analysis.

Donor cell neoplasms are rare complications of treatment regimens that involve stem cell transplantation for hematological malignancies, myelodysplastic processes, or certain genetic or metabolic disorders. We report a case of donor cell leukemia in a pediatric patient with a history of acute myeloid leukemia that manifested as recurrent AML FAB type M5 fourteen months after umbilical cord blood transplantation. Although there was some immunophenotypic drift from the patient's original AML and their posttransplant presentation, the initial pathological impression was of recurrent disease. Bone marrow engraftment analysis by multiplex PCR of short tandem repeat markers performed on the patient's diagnostic specimen showed complete engraftment by donor cells, with a loss of heterozygosity in the donor alleles on chromosome 7. This led to the reinterpretation of this patient's disease as donor-derived leukemia. This interpretation was supported by a routine karyotype and fluorescence in situ hybridization analysis showing loss of chromosome 7 and a male (donor) chromosome complement in this female patient. Also noted was a loss of the patient's presenting chromosomal abnormality, t(11;19)(q23;p13). This case highlights the need for close coordination between all aspects of clinical testing for the transplant patient, including molecular engraftment studies, when distinguishing the very common complication of recurrent disease from the exceedingly rare complication of donor cell leukemia.

Authors
Crow, J; Youens, K; Michalowski, S; Perrine, G; Emhart, C; Johnson, F; Gerling, A; Kurtzberg, J; Goodman, BK; Sebastian, S; Rehder, CW; Datto, MB
MLA Citation
Crow, J, Youens, K, Michalowski, S, Perrine, G, Emhart, C, Johnson, F, Gerling, A, Kurtzberg, J, Goodman, BK, Sebastian, S, Rehder, CW, and Datto, MB. "Donor cell leukemia in umbilical cord blood transplant patients: a case study and literature review highlighting the importance of molecular engraftment analysis." J Mol Diagn 12.4 (July 2010): 530-537. (Review)
PMID
20431036
Source
pubmed
Published In
The Journal of molecular diagnostics : JMD
Volume
12
Issue
4
Publish Date
2010
Start Page
530
End Page
537
DOI
10.2353/jmoldx.2010.090215

Utilization of genomic signatures for chemotherapy response in prospective clinical studies

Authors
Barry, W; Acharya, C; Datto, MB; Dressman, HK; Marcom, PK; Ready, N; Ginsburg, GS; Potti, A; Nevins, JR
MLA Citation
Barry, W, Acharya, C, Datto, MB, Dressman, HK, Marcom, PK, Ready, N, Ginsburg, GS, Potti, A, and Nevins, JR. "Utilization of genomic signatures for chemotherapy response in prospective clinical studies." JOURNAL OF CLINICAL ONCOLOGY 28.15 (May 20, 2010).
Source
wos-lite
Published In
Journal of Clinical Oncology
Volume
28
Issue
15
Publish Date
2010

SMAD4 is required for development of maximal endotoxin tolerance.

Initial exposure of monocytes/macrophages to LPS induces hyporesponsiveness to a second challenge with LPS, a phenomenon termed LPS tolerance. Molecular mechanisms responsible for endotoxin tolerance are not well defined. We and others have shown that IL-1R-associated kinase (IRAK)-M and SHIP-1 proteins, negative regulators of TLR4 signaling, increase in tolerized cells. TGF-beta1, an anti-inflammatory cytokine, is upregulated following LPS stimulation, mediating its effect through SMAD family proteins. Using a monocytic cell line, THP1, we show that LPS activates endogenous SMAD4, inducing its migration into the nucleus and increasing its expression. Secondary challenge with high dose LPS following initial low-dose LPS exposure does not increase IRAK-M or SHIP1 protein expression in small hairpin (sh)SMAD4 THP-1 cells compared with control shLUC THP1 cells. TNF-alpha concentrations in culture supernatants after second LPS challenge are higher in shSMAD4 THP-1 cells than shLUC THP1 cells, indicating failure to induce maximal tolerance in absence of SMAD4 signaling. Identical results are seen in primary murine macrophages and mouse embryonic fibroblasts, demonstrating the biological significance of our findings. TGF-beta1 treatment does not increase IRAK-M or SHIP1 protein expression in shSMAD4 THP-1 cells, whereas it does so in shLUC THP1 cells, indicating that TGF-beta1 regulates IRAK-M and SHIP1 expression through a SMAD4-dependent pathway. Knockdown of endogenous SHIP1 by shSHIP1 RNA decreases native and inducible IRAK-M protein expression and prevents development of endotoxin tolerance in THP1 cells. We conclude that in THP-1 cells and primary murine cells, SMAD4 signaling is required for maximal induction of endotoxin tolerance via modulation of SHIP1 and IRAK-M.

Authors
Pan, H; Ding, E; Hu, M; Lagoo, AS; Datto, MB; Lagoo-Deenadayalan, SA
MLA Citation
Pan, H, Ding, E, Hu, M, Lagoo, AS, Datto, MB, and Lagoo-Deenadayalan, SA. "SMAD4 is required for development of maximal endotoxin tolerance." J Immunol 184.10 (May 15, 2010): 5502-5509.
PMID
20404275
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
184
Issue
10
Publish Date
2010
Start Page
5502
End Page
5509
DOI
10.4049/jimmunol.0901601

Intratumor heterogeneity and precision of microarray-based predictors of breast cancer biology and clinical outcome.

PURPOSE: Identifying sources of variation in expression microarray data and the effect of variance in gene expression measurements on complex predictive and diagnostic models is essential when translating microarray-based experimental approaches into clinical assays. The technical reproducibility of microarray platforms is well established. Here, we investigate the additional impact of intratumor heterogeneity, a largely unstudied component of variance, on the performance of several microarray-based assays in breast cancer. PATIENTS AND METHODS: Genome-wide expression profiling was performed on 50 core needle biopsies from 18 breast cancer patients using Affymetrix GeneChip Human Genome U133 Plus 2.0 arrays. Global profiles of expression were characterized using unsupervised clustering methods and variance components models. Array-based measures of estrogen receptor (ER) and progesterone receptor (PR) status were compared with immunohistochemistry. The precision of genomic predictors of ER pathway status, recurrence risk, and sensitivity to chemotherapeutics was evaluated by interclass correlation. RESULTS: Global patterns of gene expression demonstrated that intratumor variation was substantially less than the total variation observed across the patient population. Nevertheless, a fraction of genes exhibited significant intratumor heterogeneity in expression. A high degree of reproducibility was observed in single-gene predictors of ER (intraclass correlation coefficient [ICC] = 0.94) and PR expression (ICC = 0.90), and in a multigene predictor of ER pathway activation (ICC = 0.98) with high concordance with immunohistochemistry. Substantial agreement was also observed for multigene signatures of cancer recurrence (ICC = 0.71) and chemotherapeutic sensitivity (ICC = 0.72 and 0.64). CONCLUSION: Intratumor heterogeneity, although present at the level of individual gene expression, does not preclude precise microarray-based predictions of tumor behavior or clinical outcome in breast cancer patients.

Authors
Barry, WT; Kernagis, DN; Dressman, HK; Griffis, RJ; Hunter, JD; Olson, JA; Marks, JR; Ginsburg, GS; Marcom, PK; Nevins, JR; Geradts, J; Datto, MB
MLA Citation
Barry, WT, Kernagis, DN, Dressman, HK, Griffis, RJ, Hunter, JD, Olson, JA, Marks, JR, Ginsburg, GS, Marcom, PK, Nevins, JR, Geradts, J, and Datto, MB. "Intratumor heterogeneity and precision of microarray-based predictors of breast cancer biology and clinical outcome." J Clin Oncol 28.13 (May 1, 2010): 2198-2206.
PMID
20368555
Source
pubmed
Published In
Journal of Clinical Oncology
Volume
28
Issue
13
Publish Date
2010
Start Page
2198
End Page
2206
DOI
10.1200/JCO.2009.26.7245

A pathway-based classification of human breast cancer.

The hallmark of human cancer is heterogeneity, reflecting the complexity and variability of the vast array of somatic mutations acquired during oncogenesis. An ability to dissect this heterogeneity, to identify subgroups that represent common mechanisms of disease, will be critical to understanding the complexities of genetic alterations and to provide a framework to develop rational therapeutic strategies. Here, we describe a classification scheme for human breast cancer making use of patterns of pathway activity to build on previous subtype characterizations using intrinsic gene expression signatures, to provide a functional interpretation of the gene expression data that can be linked to therapeutic options. We show that the identified subgroups provide a robust mechanism for classifying independent samples, identifying tumors that share patterns of pathway activity and exhibit similar clinical and biological properties, including distinct patterns of chromosomal alterations that were not evident in the heterogeneous total population of tumors. We propose that this classification scheme provides a basis for understanding the complex mechanisms of oncogenesis that give rise to these tumors and to identify rational opportunities for combination therapies.

Authors
Gatza, ML; Lucas, JE; Barry, WT; Kim, JW; Wang, Q; Crawford, MD; Datto, MB; Kelley, M; Mathey-Prevot, B; Potti, A; Nevins, JR
MLA Citation
Gatza, ML, Lucas, JE, Barry, WT, Kim, JW, Wang, Q, Crawford, MD, Datto, MB, Kelley, M, Mathey-Prevot, B, Potti, A, and Nevins, JR. "A pathway-based classification of human breast cancer." Proc Natl Acad Sci U S A 107.15 (April 13, 2010): 6994-6999.
PMID
20335537
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
107
Issue
15
Publish Date
2010
Start Page
6994
End Page
6999
DOI
10.1073/pnas.0912708107

NDRG4 is required for cell cycle progression and survival in glioblastoma cells.

NDRG4 is a largely unstudied member of the predominantly tumor suppressive N-Myc downstream-regulated gene (NDRG) family. Unlike its family members NDRG1-3, which are ubiquitously expressed, NDRG4 is expressed almost exclusively in the heart and brain. Given this tissue-specific expression pattern and the established tumor suppressive roles of the NDRG family in regulating cellular proliferation, we investigated the cellular and biochemical functions of NDRG4 in the context of astrocytes and glioblastoma multiforme (GBM) cells. We show that, in contrast to NDRG2, NDRG4 expression is elevated in GBM and NDRG4 is required for the viability of primary astrocytes, established GBM cell lines, and both CD133(+) (cancer stem cell (CSC)-enriched) and CD133(-) primary GBM xenograft cells. While NDRG4 overexpression has no effect on cell viability, NDRG4 knockdown causes G(1) cell cycle arrest followed by apoptosis. The initial G(1) arrest is associated with a decrease in cyclin D1 expression and an increase in p27(Kip1) expression, and the subsequent apoptosis is associated with a decrease in the expression of XIAP and survivin. As a result of these effects on cell cycle progression and survival, NDRG4 knockdown decreases the tumorigenic capacity of established GBM cell lines and GBM CSC-enriched cells that have been implanted intracranially into immunocompromised mice. Collectively, these data indicate that NDRG4 is required for cell cycle progression and survival, thereby diverging in function from its tumor suppressive family member NDRG2 in astrocytes and GBM cells.

Authors
Schilling, SH; Hjelmeland, AB; Radiloff, DR; Liu, IM; Wakeman, TP; Fielhauer, JR; Foster, EH; Lathia, JD; Rich, JN; Wang, X-F; Datto, MB
MLA Citation
Schilling, SH, Hjelmeland, AB, Radiloff, DR, Liu, IM, Wakeman, TP, Fielhauer, JR, Foster, EH, Lathia, JD, Rich, JN, Wang, X-F, and Datto, MB. "NDRG4 is required for cell cycle progression and survival in glioblastoma cells." J Biol Chem 284.37 (September 11, 2009): 25160-25169.
PMID
19592488
Source
pubmed
Published In
The Journal of biological chemistry
Volume
284
Issue
37
Publish Date
2009
Start Page
25160
End Page
25169
DOI
10.1074/jbc.M109.012484

Chordoid glioma: a case report and molecular characterization of five cases.

Chordoid gliomas are rare, slow-growing neoplasms of the anterior third ventricle. We reported a case of chordoid glioma in a 41-year-old man with obstructive hydrocephalus. Histologically, the tumor consisted of polygonal epithelioid cells admixed with elongated cells in a myxoid stroma. A prominent lymphoplasmacytic infiltrate was present. The tumor cells expressed glial fibrillary acidic protein (GFAP), epithelial membrane antigen (EMA), vimentin, CD31, CD34, epidermal growth factor receptor (EGFR) and S100 but were negative for pankeratin and E-cadherin. The percentage of Ki67 positive cells was approximately 3%. Weak p53 immunoreactivity was seen in less than 10% of the cells. Array comparative genomic hybridization performed on this case, as well as on four other archived cases, showed losses at several loci. Fluorescence in situ hybridization (FISH) confirmed consistent genetic alterations at 9p21 and 11q13. These are the fifth through ninth reported cases of chordoid gliomas with molecular characterization suggesting a distinct genetic origin from other gliomas.

Authors
Horbinski, C; Dacic, S; McLendon, RE; Cieply, K; Datto, M; Brat, DJ; Chu, CT
MLA Citation
Horbinski, C, Dacic, S, McLendon, RE, Cieply, K, Datto, M, Brat, DJ, and Chu, CT. "Chordoid glioma: a case report and molecular characterization of five cases." Brain Pathol 19.3 (July 2009): 439-448.
PMID
18652591
Source
pubmed
Published In
Brain Pathology
Volume
19
Issue
3
Publish Date
2009
Start Page
439
End Page
448
DOI
10.1111/j.1750-3639.2008.00196.x

Deletion of the protein kinase A/protein kinase G target SMTNL1 promotes an exercise-adapted phenotype in vascular smooth muscle.

In vivo protein kinases A and G (PKA and PKG) coordinately phosphorylate a broad range of substrates to mediate their various physiological effects. The functions of many of these substrates have yet to be defined genetically. Herein we show a role for smoothelin-like protein 1 (SMTNL1), a novel in vivo target of PKG/PKA, in mediating vascular adaptations to exercise. Aortas from smtnl1(-/-) mice exhibited strikingly enhanced vasorelaxation before exercise, similar in extent to that achieved after endurance training of wild-type littermates. Additionally, contractile responses to alpha-adrenergic agonists were greatly attenuated. Immunological studies showed SMTNL1 is expressed in smooth muscle and type 2a striated muscle fibers. Consistent with a role in adaptations to exercise, smtnl1(-/-) mice also exhibited increased type 2a fibers before training and better performance after forced endurance training compared smtnl1(+/+) mice. Furthermore, exercise was found to reduce expression of SMTNL1, particularly in female mice. In both muscle types, SMTNL1 is phosphorylated at Ser-301 in response to adrenergic signals. In vitro SMTNL1 suppresses myosin phosphatase activity through a substrate-directed effect, which is relieved by Ser-301 phosphorylation. Our findings suggest roles for SMTNL1 in cGMP/cAMP-mediated adaptations to exercise through mechanisms involving direct modulation of contractile activity.

Authors
Wooldridge, AA; Fortner, CN; Lontay, B; Akimoto, T; Neppl, RL; Facemire, C; Datto, MB; Kwon, A; McCook, E; Li, P; Wang, S; Thresher, RJ; Miller, SE; Perriard, J-C; Gavin, TP; Hickner, RC; Coffman, TM; Somlyo, AV; Yan, Z; Haystead, TAJ
MLA Citation
Wooldridge, AA, Fortner, CN, Lontay, B, Akimoto, T, Neppl, RL, Facemire, C, Datto, MB, Kwon, A, McCook, E, Li, P, Wang, S, Thresher, RJ, Miller, SE, Perriard, J-C, Gavin, TP, Hickner, RC, Coffman, TM, Somlyo, AV, Yan, Z, and Haystead, TAJ. "Deletion of the protein kinase A/protein kinase G target SMTNL1 promotes an exercise-adapted phenotype in vascular smooth muscle." J Biol Chem 283.17 (April 25, 2008): 11850-11859.
PMID
18310078
Source
pubmed
Published In
The Journal of biological chemistry
Volume
283
Issue
17
Publish Date
2008
Start Page
11850
End Page
11859
DOI
10.1074/jbc.M708628200

Extracellular matrix protein betaig-h3/TGFBI promotes metastasis of colon cancer by enhancing cell extravasation.

Metastasis, the major cause of cancer death, is a multistep process that requires interactions between cancer cells and stromal cells and between cancer cells and extracellular matrix. Molecular alterations of the extracellular matrix in the tumor microenvironment have a considerable impact on the metastatic process during tumorigenesis. Here we report that elevated expression of betaig-h3/TGFBI (transforming growth factor, beta-induced), an extracellular matrix protein secreted by colon cancer cells, is associated with high-grade human colon cancers. Ectopic expression of the betaig-h3 protein enhanced the aggressiveness and altered the metastatic properties of colon cancer cells in vivo. Inhibition of betaig-h3 expression dramatically reduced metastasis. Mechanistically, betaig-h3 appears to promote extravasation, a critical step in the metastatic dissemination of cancer cells, by inducing the dissociation of VE-cadherin junctions between endothelial cells via activation of the integrin alphavbeta5-Src signaling pathway. Thus, cancers associated with overexpression of betaig-h3 may have an increased metastatic potential, leading to poor prognosis in cancer patients.

Authors
Ma, C; Rong, Y; Radiloff, DR; Datto, MB; Centeno, B; Bao, S; Cheng, AWM; Lin, F; Jiang, S; Yeatman, TJ; Wang, X-F
MLA Citation
Ma, C, Rong, Y, Radiloff, DR, Datto, MB, Centeno, B, Bao, S, Cheng, AWM, Lin, F, Jiang, S, Yeatman, TJ, and Wang, X-F. "Extracellular matrix protein betaig-h3/TGFBI promotes metastasis of colon cancer by enhancing cell extravasation." Genes Dev 22.3 (February 1, 2008): 308-321.
PMID
18245446
Source
pubmed
Published In
Genes & development
Volume
22
Issue
3
Publish Date
2008
Start Page
308
End Page
321
DOI
10.1101/gad.1632008

PITK, a PP1 targeting subunit that modulates the phosphorylation of the transcriptional regulator hnRNP K.

Protein phosphatase-1 (PP1), through interactions with substrate targeting subunits, plays critical roles in the regulation of numerous cellular processes. Herein, we describe a newly identified regulatory subunit (PITK; Phosphatase Interactor Targeting K protein) that specifically targets the catalytic subunit of PP1 to nuclear foci to selectively bind and dephosphorylate the transcriptional regulator heterogeneous nuclear ribonucleoprotein K (hnRNP K) at a regulatory S284 site. Additionally, PITK is phosphorylated in vivo at S1013 and S1017, residues that flank or reside within the PP1C-binding motif, and this phosphorylation negatively regulates the binding of the phosphatase to PITK. A mutant variant, S1013,1017A-PITK, when expressed in intact cells, exhibited an increase in native PP1 binding and elicited a more profound dephosphorylation of hnRNPK at S284. A global analysis of transcription by Affymetrix microarray revealed that the expression of PITK resulted in the altered expression of 47 genes, including a marked induction of MEK5 (>14-fold, p<0.007). Additionally, the effects of PITK and S1013,1017A-PITK on transcription could be modulated by the co-expression of hnRNP K. Taken together, our findings provide a putative mechanism by which transcriptional activity of hnRNP K can be discretely controlled through the regulation of PP1 activity.

Authors
Kwiek, NC; Thacker, DF; Datto, MB; Megosh, HB; Haystead, TAJ
MLA Citation
Kwiek, NC, Thacker, DF, Datto, MB, Megosh, HB, and Haystead, TAJ. "PITK, a PP1 targeting subunit that modulates the phosphorylation of the transcriptional regulator hnRNP K." Cell Signal 18.10 (October 2006): 1769-1778.
PMID
16564677
Source
pubmed
Published In
Cellular Signalling
Volume
18
Issue
10
Publish Date
2006
Start Page
1769
End Page
1778
DOI
10.1016/j.cellsig.2006.01.019

A phosphatase controls the fate of receptor-regulated Smads.

In this issue of Cell, Lin et al. (2006) answer one of the long-standing questions in the TGFbeta field by identifying a phosphatase, PPM1A, that directly dephosphorylates Smad2 and Smad3 to limit their activation.

Authors
Schilling, SH; Datto, MB; Wang, X-F
MLA Citation
Schilling, SH, Datto, MB, and Wang, X-F. "A phosphatase controls the fate of receptor-regulated Smads." Cell 125.5 (June 2, 2006): 838-840. (Review)
PMID
16751094
Source
pubmed
Published In
Cell
Volume
125
Issue
5
Publish Date
2006
Start Page
838
End Page
840
DOI
10.1016/j.cell.2006.05.015

Synchronous endometrial and ovarian tumors: metastatic disease or independent primaries?

Authors
Robboy, SJ; Datto, MB
MLA Citation
Robboy, SJ, and Datto, MB. "Synchronous endometrial and ovarian tumors: metastatic disease or independent primaries?." Hum Pathol 36.6 (June 2005): 597-599.
PMID
16021564
Source
pubmed
Published In
Human Pathology
Volume
36
Issue
6
Publish Date
2005
Start Page
597
End Page
599
DOI
10.1016/j.humpath.2005.05.012

Ubiquitin-mediated degradation a mechanism for fine-tuning TGF-beta signaling.

Effects of the cytokine TGF-beta can be dampened by E3 ubiquitin ligases that target specific Smads, the TGF-beta signal transducers, for proteolytic destruction. Two papers in this issue of Cell highlight the importance of this mechanism in regulating the in vivo effects of TGF-beta. The first paper identifies and characterizes a novel Smad4 ubiquitin ligase, and the second paper redefines the role of a previously identified Smad1 ubiquitin ligase, Smurf-1 (Dupont et al., 2005; Yamashita et al., 2005).

Authors
Datto, M; Wang, X-F
MLA Citation
Datto, M, and Wang, X-F. "Ubiquitin-mediated degradation a mechanism for fine-tuning TGF-beta signaling." Cell 121.1 (April 8, 2005): 2-4.
PMID
15820671
Source
pubmed
Published In
Cell
Volume
121
Issue
1
Publish Date
2005
Start Page
2
End Page
4
DOI
10.1016/j.cell.2005.03.017

The proteome

Authors
Datto, MB; Haystead, TAJ
MLA Citation
Datto, MB, and Haystead, TAJ. "The proteome." (December 20, 2004): 153-178. (Chapter)
Source
scopus
Publish Date
2004
Start Page
153
End Page
178

Chemotherapy-induced toxic leukoencephalopathy causes a wide range of symptoms: a series of four autopsies.

We have observed an increasing number of autopsies on patients with chemotherapy-related complications. One complication is toxic leukoencephalopathy, which is due to a direct toxic effect of chemotherapeutic agents on the central nervous system white matter. Autopsies of four cases of toxic leukoencephalopathy were performed following standard protocols. The brain and spinal cord were examined routinely, and histological sections were taken for evaluation. We report here three patients with hematologic malignancies and one patient with metastatic carcinoma with chemotherapy-induced leukoencephalopathy. The first was a 56-year-old male treated with multiple chemotherapeutics for multiple myeloma. He presented with confusion and focal seizures with a rapid progression to coma and decerebrate posturing. The second was a 36-year-old male who developed mental status changes, ataxia and dysarthria following treatment for lymphoma. The third was a 16-year-old male who developed a profound peripheral and central neuropathy after chemotherapy treatment for T-cell acute lymphoblastic leukemia. The fourth was a 49-year-old female patient who was treated with multiple chemotherapeutics for Stage II breast carcinoma and subsequently developed visual acuity and field defects. The neuropathologic findings in these cases, although similar, varied in severity and distribution. The white matter was affected by severe myelin pallor, edema, and a prominent macrophage infiltrate in each of the cases. The location and extent of the central nervous system pathology correlated with the type and severity of clinical symptoms. These four cases, with their varied presenting symptoms, clinical courses, and degree of pathology, emphasize the importance of considering toxic leukoencephalopathy as an etiology of acute neurologic deterioration following high-dose chemotherapy.

Authors
Moore-Maxwell, CA; Datto, MB; Hulette, CM
MLA Citation
Moore-Maxwell, CA, Datto, MB, and Hulette, CM. "Chemotherapy-induced toxic leukoencephalopathy causes a wide range of symptoms: a series of four autopsies." Mod Pathol 17.2 (February 2004): 241-247.
PMID
14704718
Source
pubmed
Published In
Modern Pathology
Volume
17
Issue
2
Publish Date
2004
Start Page
241
End Page
247
DOI
10.1038/modpathol.3800049

Smad3 deficiency attenuates bleomycin-induced pulmonary fibrosis in mice.

Transforming growth factor-beta (TGF-beta) signaling plays an important regulatory role during lung fibrogenesis. Smad3 was identified in the pathway for transducing TGF-beta signals from the cell membrane to the nucleus. Using mice without Smad3 gene expression, we investigated whether Smad3 could regulate bleomycin-induced pulmonary fibrosis in vivo. Mice deficient in Smad3 demonstrated suppressed type I procollagen mRNA expression and reduced hydroxyproline content in the lungs compared with wild-type mice treated with bleomycin. Furthermore, loss of Smad3 greatly attenuated morphological fibrotic responses to bleomycin in the mouse lungs, suggesting that Smad3 is implicated in the pathogenesis of pulmonary fibrosis. These results show that Smad3 contributes to bleomycin-induced lung injury and that Smad3 may serve as a novel target for potential therapeutic treatment of lung fibrosis.

Authors
Zhao, J; Shi, W; Wang, Y-L; Chen, H; Bringas, P; Datto, MB; Frederick, JP; Wang, X-F; Warburton, D
MLA Citation
Zhao, J, Shi, W, Wang, Y-L, Chen, H, Bringas, P, Datto, MB, Frederick, JP, Wang, X-F, and Warburton, D. "Smad3 deficiency attenuates bleomycin-induced pulmonary fibrosis in mice." American journal of physiology. Lung cellular and molecular physiology 282.3 (March 2002): L585-L593.
PMID
11839555
Source
epmc
Published In
American journal of physiology. Lung cellular and molecular physiology
Volume
282
Issue
3
Publish Date
2002
Start Page
L585
End Page
L593
DOI
10.1152/ajplung.00151.2001

Chemotherapy induced leukoencephalopathy causes a wide range of symptoms: A series of three autopsies

Authors
Datto, MB; Moore-Maxwell, CA; Hulette, C
MLA Citation
Datto, MB, Moore-Maxwell, CA, and Hulette, C. "Chemotherapy induced leukoencephalopathy causes a wide range of symptoms: A series of three autopsies." MODERN PATHOLOGY 15.1 (January 2002): 5A-5A.
Source
wos-lite
Published In
Modern Pathology
Volume
15
Issue
1
Publish Date
2002
Start Page
5A
End Page
5A

Chemotherapy induced leukoencephalopathy causes a wide range of symptoms: A series of three autopsies

Authors
Datto, MB; Moore-Maxwell, CA; Hulette, C
MLA Citation
Datto, MB, Moore-Maxwell, CA, and Hulette, C. "Chemotherapy induced leukoencephalopathy causes a wide range of symptoms: A series of three autopsies." LABORATORY INVESTIGATION 82.1 (January 2002): 5A-5A.
Source
wos-lite
Published In
Laboratory Investigation
Volume
82
Issue
1
Publish Date
2002
Start Page
5A
End Page
5A

Smad3 deficiency attenuates bleomycin-induced pulmonary fibrosis in mice

Transforming growth factor-β (TGF-β) signaling plays an important regulatory role during lung fibrogenesis. Smad3 was identified in the pathway for transducing TGF-β signals from the cell membrane to the nucleus. Using mice without Smad3 gene expression, we investigated whether Smad3 could regulate bleomycin-induced pulmonary fibrosis in vivo. Mice deficient in Smad3 demonstrated suppressed type I procollagen mRNA expression and reduced hydroxyproline content in the lungs compared with wild-type mice treated with bleomycin. Furthermore, loss of Smad3 greatly attenuated morphological fibrotic responses to bleomycin in the mouse lungs, suggesting that Smad3 is implicated in the pathogenesis of pulmonary fibrosis. These results show that Smad3 contributes to bleomycin-induced lung injury and that Smad3 may serve as a novel target for potential therapeutic treatment of lung fibrosis.

Authors
Zhao, J; Shi, W; Wang, Y-L; Chen, H; Jr, PB; Datto, MB; Frederick, JP; Wang, X-F; Warburton, D
MLA Citation
Zhao, J, Shi, W, Wang, Y-L, Chen, H, Jr, PB, Datto, MB, Frederick, JP, Wang, X-F, and Warburton, D. "Smad3 deficiency attenuates bleomycin-induced pulmonary fibrosis in mice." American Journal of Physiology - Lung Cellular and Molecular Physiology 282.3 26-3 (2002): L585-L593.
Source
scival
Published In
American journal of physiology. Lung cellular and molecular physiology
Volume
282
Issue
3 26-3
Publish Date
2002
Start Page
L585
End Page
L593

The loss of Smad3 results in a lower rate of bone formation and osteopenia through dysregulation of osteoblast differentiation and apoptosis.

Smad3 is a well-characterized intracellular effector of the transforming growth factor beta (TGF-beta) signaling pathway and was implicated recently in the potentiation of vitamin D receptor (VDR)-mediated signaling. Given that both TGF-beta and vitamin D are important regulators of bone remodeling, it is expected that Smad3 plays an integral role in normal maintenance of bone. However, the exact mechanisms by which Smad3 functions in bone remodeling are unknown. Here, we show that mice with targeted deletion of Smad3 are osteopenic with less cortical and cancellous bone compared with wild-type littermates. Decreases in bone mineral density (BMD) in Smad3 null mice reflect the inability of osteoblasts to balance osteoclast activity, although osteoclast numbers are normal and vitamin D mediated serum calcium homeostasis is maintained. The osteopenia of Smad3 null mice is attributed to a decreased rate of bone formation associated with increased osteocyte number and apoptosis. These findings are supported by studies with isolated primary osteoblasts that show TGF-beta can no longer inhibit the differentiation of osteoblasts in the absence of Smad3; yet, TGF-beta-stimulated proliferation remains intact. Together these data support a model that a loss of Smad3 increases the osteocyte fate of the osteoblast and decreases the duration of osteoblast function by shortening lifespan, ultimately resulting in osteopenia.

Authors
Borton, AJ; Frederick, JP; Datto, MB; Wang, XF; Weinstein, RS
MLA Citation
Borton, AJ, Frederick, JP, Datto, MB, Wang, XF, and Weinstein, RS. "The loss of Smad3 results in a lower rate of bone formation and osteopenia through dysregulation of osteoblast differentiation and apoptosis." J Bone Miner Res 16.10 (October 2001): 1754-1764.
PMID
11585338
Source
pubmed
Published In
Journal of Bone and Mineral Research
Volume
16
Issue
10
Publish Date
2001
Start Page
1754
End Page
1764
DOI
10.1359/jbmr.2001.16.10.1754

RB regulates transcription of the p21/WAF1/CIP1 gene.

We have previously shown that RB plays an important role in the maintenance of the epithelial phenotype. p21 is also involved in several terminal differentiation systems including keratinocytes. We report here that p21 is an RB target gene in epithelial cells, but not in fibroblasts where RB is unable to transactivate p21 transcriptional expression. In epithelial cells, when RB family factors were inactivated by SV40 T antigen (LT), p21 expression was strongly repressed, whereas its expression was not affected when the cells were transformed by a mutated LT leaving RB active but inactivating p53. Moreover, retransformation by RB of LT transformed epithelial cells totally restored p21 expression. By cotransfection experiments and using deletions and point mutations of the p21 promoter, we show that the minimal region required for the RB-mediated transcriptional activation maps to a GC-rich region located between -83 and -74. This region is shown to interact specifically with the transcription factor Sp1 and Sp3. Thus for the first time, we show a positive transcriptional relationship between RB and p21 in epithelial cells. Since p21 keeps RB in a hypophosphorylated state important for its transcriptional activity during differentiation, our results imply an auto-loop of regulation between RB and p21 that may be essential for the maintenance of the differentiation state. We propose that this transcriptional relationship might be necessary of their roles in cell cycle arrest and in several differentiation pathways.

Authors
Decesse, JT; Medjkane, S; Datto, MB; Crémisi, CE
MLA Citation
Decesse, JT, Medjkane, S, Datto, MB, and Crémisi, CE. "RB regulates transcription of the p21/WAF1/CIP1 gene." Oncogene 20.8 (February 22, 2001): 962-971.
PMID
11314031
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
20
Issue
8
Publish Date
2001
Start Page
962
End Page
971
DOI
10.1038/sj.onc.1204169

An essential role for Smad3 in bone formation through regulation of osteoblast differentiation.

Authors
Borton, AJ; Frederick, JP; Datto, MB; Wang, X; Weinstein, RS
MLA Citation
Borton, AJ, Frederick, JP, Datto, MB, Wang, X, and Weinstein, RS. "An essential role for Smad3 in bone formation through regulation of osteoblast differentiation." JOURNAL OF BONE AND MINERAL RESEARCH 15 (September 2000): S189-S189.
Source
wos-lite
Published In
Journal of Bone and Mineral Research
Volume
15
Publish Date
2000
Start Page
S189
End Page
S189

The Smads: transcriptional regulation and mouse models.

The field of transforming growth factor-beta (TGF-beta) signaling sees periodic discoveries that revolutionize our thinking, redirect our experiments, and peak our excitement. One of the first such discoveries was less than a decade ago: the molecular cloning of the type I and type II TGF-beta receptors. This breakthrough defined a novel family of serine/threonine kinase receptors, which led to the description of an ever-expanding superfamily. The discovery of how these receptors are grouped on the cell surface, bind TGF-beta and are activated by specific phosphorylation events further defined the uniqueness of this system in comparison to other families of growth factor receptors. Now, once again, the TGF-beta field has been revolutionized. This time, the discovery is the Smad family of proteins. Although one can hardly imagine TGF-beta without the Smads, the cloning of the Smads and their implication in TGF-beta signaling was only four years ago. Since that time, great advances have been made in our understanding of the Smads as transcription factors, which are activated by receptor mediated phosphorylation. In addition, animal models for a loss of Smad function have provided insight into the role of specific Smads in a variety of physiologic systems. The Smad field has been growing exponentially. A comprehensive review of all aspects of the Smads, therefore, would be beyond the scope of a single review. Instead, this review highlights some of the general aspects of Smad function, and then focuses on the role of specific Smad family members in transcriptional regulation, animal physiology, and disease processes.

Authors
Datto, M; Wang, XF
MLA Citation
Datto, M, and Wang, XF. "The Smads: transcriptional regulation and mouse models." Cytokine Growth Factor Rev 11.1-2 (March 2000): 37-48. (Review)
PMID
10708951
Source
pubmed
Published In
Cytokine & Growth Factor Reviews
Volume
11
Issue
1-2
Publish Date
2000
Start Page
37
End Page
48

Transforming growth factor-beta-mediated p15(INK4B) induction and growth inhibition in astrocytes is SMAD3-dependent and a pathway prominently altered in human glioma cell lines.

We sought to characterize the pathway by which the multifunctional cytokine transforming growth factor-beta (TGF-beta) inhibits the proliferation of normal astrocytes, and we analyzed the alterations in the TGF-beta pathway in human glioma cell lines. Upon TGF-beta treatment, primary rat astrocytes showed a significant decrease in DNA synthesis upon thymidine incorporation with a cell cycle arrest in the G(1) phase. Western analysis of the astrocytes revealed that the expression of the cyclin-dependent kinase inhibitor (CdkI) p15(INK4B) was significantly up-regulated upon TGF-beta treatment without a change in other CdkI levels. The retinoblastoma protein (Rb) became hypophosphorylated, and Cdk2 activity decreased. Analysis of Smad3 null mouse astrocytes showed a significant loss of both TGF-beta-mediated growth inhibition and p15(INK4B) induction compared with wild-type mouse astrocytes. Infection of rat astrocytes by SMAD3 and SMAD4 adenoviruses failed to induce increased expression of p15(INK4B), implying indirect transcriptional regulation of p15(INK4B) by SMAD3. High-grade human gliomas secrete TGF-beta, yet are resistant to its growth inhibitory effects. Analysis of the effects of TGF-beta on 12 human glioma cell lines showed that TGF-beta mildly inhibited the growth of six lines, had no effect on four lines, and stimulated the growth of two lines. The majority of glioma lines had homozygous deletions of the p15(INK4B) gene, except for two lines that expressed p15(INK4B) protein, which was induced further upon TGF-beta treatment. Three lines mildly induced CdkI p21(WAF1) expression in response to TGF-beta. Most tumor lines retained other TGF-beta-mediated responses, including extracellular matrix protein and angiogenic factor secretion, which may contribute to increased malignant behavior. This suggests that the loss of p15(INK4B) may explain, in part, the selective loss of growth inhibition by TGF-beta in gliomas to form a more aggressive tumor phenotype.

Authors
Rich, JN; Zhang, M; Datto, MB; Bigner, DD; Wang, XF
MLA Citation
Rich, JN, Zhang, M, Datto, MB, Bigner, DD, and Wang, XF. "Transforming growth factor-beta-mediated p15(INK4B) induction and growth inhibition in astrocytes is SMAD3-dependent and a pathway prominently altered in human glioma cell lines." J Biol Chem 274.49 (December 3, 1999): 35053-35058.
PMID
10574984
Source
pubmed
Published In
The Journal of biological chemistry
Volume
274
Issue
49
Publish Date
1999
Start Page
35053
End Page
35058

Ras induces p21Cip1/Waf1 cyclin kinase inhibitor transcriptionally through Sp1-binding sites.

p21Cip1/Waf1 cyclin-dependent kinase inhibitor (p21) is inducible by Raf and mitogen-activated protein kinase kinase (MAPKK), but the level of regulation is unknown. We show here by conditional and transient Ras-expression models that Ras induces p21. Induction of p21 in conditionally Ras-expressing cells is posttranscriptional utilizing mitogen-activated protein kinase (MAPK) pathway. Transient, high-level Ras-expression induces transcriptional activation of p21 mediated by a GC-rich region in p21 promoter -83-54 bp relative to the transcription initiation site containing binding sites for Sp1-family transcription factors. Mutation of either Sp1-binding site 2 or 4 in this region decreases the magnitude of induction of promoter activity by Ras, but only the simultaneous mutation of both sites abolishes fully the induction. Electrophoretic mobility shift assays using an oligonucleotide corresponding to Sp1-binding site 2 indicate that both Sp1 and Sp3 transcription factors bind to this region. The results demonstrate that the central cytosolic growth regulator Ras is a potent transcriptional and posttranscriptional inducer of the nuclear growth inhibitor p21.

Authors
Kivinen, L; Tsubari, M; Haapajärvi, T; Datto, MB; Wang, XF; Laiho, M
MLA Citation
Kivinen, L, Tsubari, M, Haapajärvi, T, Datto, MB, Wang, XF, and Laiho, M. "Ras induces p21Cip1/Waf1 cyclin kinase inhibitor transcriptionally through Sp1-binding sites." Oncogene 18.46 (November 4, 1999): 6252-6261.
PMID
10597223
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
18
Issue
46
Publish Date
1999
Start Page
6252
End Page
6261
DOI
10.1038/sj.onc.1203000

Cooperation of Sp1 and p300 in the induction of the CDK inhibitor p21WAF1/CIP1 during NGF-mediated neuronal differentiation.

Addition of nerve growth factor (NGF) to PC12 cells promotes neuronal differentiation while inhibiting cell proliferation. In order to understand how NGF exerts its antimitogenic effect during differentiation, we have studied the mechanism by which this factor activates the promoter of the CDK inhibitor p21W4F1/CIP1. The minimal region of the p21 promoter required for the NGF-induction was mapped to a contiguous stretch of 10 bp located 83 bases upstream of the transcription initiation site. This GC-rich region was shown to interact specifically with the transcription factor Sp1 and the related protein Sp3, in either exponentially-growing or NGF-treated PC12 cells. The addition of NGF resulted in an accumulation of the transcriptional co-activator p300 in complexes associated with the NGF-responsive region. Transcriptional activity of Sp1, Sp3 and p300 was specifically induced by NGF in a Gal4-fusion assay, indicating that induction of p21 during neuronal differentiation may involve regulation of the activity of these factors by NGF. Furthermore, p300 was able to act as a co-activator for Sp1-mediated transcriptional activation in PC12 cells, suggesting that p300 and Sp1 may cooperate in activating p21 transcription during the withdrawal of neuronal precursors from the cell cycle. This hypothesis is supported by experiments showing that p300 and Sp1 form complexes in PC12 cells.

Authors
Billon, N; Carlisi, D; Datto, MB; van Grunsven, LA; Watt, A; Wang, XF; Rudkin, BB
MLA Citation
Billon, N, Carlisi, D, Datto, MB, van Grunsven, LA, Watt, A, Wang, XF, and Rudkin, BB. "Cooperation of Sp1 and p300 in the induction of the CDK inhibitor p21WAF1/CIP1 during NGF-mediated neuronal differentiation." Oncogene 18.18 (May 6, 1999): 2872-2882.
PMID
10362258
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
18
Issue
18
Publish Date
1999
Start Page
2872
End Page
2882
DOI
10.1038/sj.onc.1202712

Smads bind directly to the Jun family of AP-1 transcription factors.

Smad3 and Smad4 are sequence-specific DNA-binding factors that bind to their consensus DNA-binding sites in response to transforming growth factor beta (TGFbeta) and activate transcription. Recent evidence implicates Smad3 and Smad4 in the transcriptional activation of consensus AP-1 DNA-binding sites that do not interact with Smads directly. Here, we report that Smad3 and Smad4 can physically interact with AP-1 family members. In vitro binding studies demonstrate that both Smad3 and Smad4 bind all three Jun family members: JunB, cJun, and JunD. The Smad interacting region of JunB maps to a C-terminal 20-amino acid sequence that is partially conserved in cJun and JunD. We show that Smad3 and Smad4 also associate with an endogenous form of cJun that is rapidly phosphorylated in response to TGFbeta. Providing evidence for the importance of this interaction between Smad and Jun proteins, we demonstrate that Smad3 is required for the activation of concatamerized AP-1 sites in a reporter construct that has previously been characterized as unable to bind Smad proteins directly. Together, these data suggest that TGFbeta-mediated transcriptional activation through AP-1 sites may involve a regulated interaction between Smads and AP-1 transcription factors.

Authors
Liberati, NT; Datto, MB; Frederick, JP; Shen, X; Wong, C; Rougier-Chapman, EM; Wang, XF
MLA Citation
Liberati, NT, Datto, MB, Frederick, JP, Shen, X, Wong, C, Rougier-Chapman, EM, and Wang, XF. "Smads bind directly to the Jun family of AP-1 transcription factors." Proc Natl Acad Sci U S A 96.9 (April 27, 1999): 4844-4849.
PMID
10220381
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
96
Issue
9
Publish Date
1999
Start Page
4844
End Page
4849

UV radiation is a transcriptional inducer of p21(Cip1/Waf1) cyclin-kinase inhibitor in a p53-independent manner.

p53 target genes p21(Cip1/Waf1) cyclin-kinase inhibitor (p21 CKI), GADD45, bax, and cyclin G and genes affecting the redox state of the cells are implicated in p53 damage control responses. In order to attribute their functions and dependency of p53 in UV-damaged cells we undertook an analysis of UVC responses of fibroblasts derived from p53 knock-out mice. UVC radiation efficiently and rapidly inhibited DNA replication in both p53 -/- and +/+ cells. The arrest was persistent in p53 -/- fibroblasts and cells underwent apoptosis, whereas p53 +/+ cells recovered and reentered the cycle. Protein and mRNA analyses of p21 expression showed that it was induced up to sixfold with similar kinetics both in the presence and in the absence of p53. However, high doses of UV abrogated the p21 response in p53 -/- cells, whereas it was maintained in cells with normal p53. UVC radiation transcriptionally activated p21 expression as demonstrated by luciferase reporter assays using deletion constructs of the p21 promoter. The promoter assays further confirmed the independency of p53-binding sites in the activation and linked UV-responsive transcriptional regulation of p21 to two Sp1 consensus binding sites within -61 bp of the transcription initiation site. A weaker regulation was mediated by elements between -1300 to -500 bp relative to the transcription initiation site. The results suggest that in fibroblasts UVC radiation is a rapid and efficient inducer of p21 expression also in a p53-independent manner.

Authors
Haapajärvi, T; Kivinen, L; Heiskanen, A; des Bordes, C; Datto, MB; Wang, XF; Laiho, M
MLA Citation
Haapajärvi, T, Kivinen, L, Heiskanen, A, des Bordes, C, Datto, MB, Wang, XF, and Laiho, M. "UV radiation is a transcriptional inducer of p21(Cip1/Waf1) cyclin-kinase inhibitor in a p53-independent manner." Exp Cell Res 248.1 (April 10, 1999): 272-279.
PMID
10094833
Source
pubmed
Published In
Experimental Cell Research
Volume
248
Issue
1
Publish Date
1999
Start Page
272
End Page
279
DOI
10.1006/excr.1999.4403

Targeted disruption of Smad3 reveals an essential role in transforming growth factor beta-mediated signal transduction.

The Smads are a family of nine related proteins which function as signaling intermediates for the transforming growth factor beta (TGF-beta) superfamily of ligands. To discern the in vivo functions of one of these Smads, Smad3, we generated mice harboring a targeted disruption of this gene. Smad3 null mice, although smaller than wild-type littermates, are viable, survive to adulthood, and exhibit an early phenotype of forelimb malformation. To study the cellular functions of Smad3, we generated Smad3 null mouse embryonic fibroblasts (MEFs) and dermal fibroblasts. We demonstrate that null MEFs have lost the ability to form Smad-containing DNA binding complexes and are unable to induce transcription from the TGF-beta-responsive promoter construct, p3TP-lux. Using the primary dermal fibroblasts, we also demonstrate that Smad3 is integral for induction of endogenous plasminogen activator inhibitor 1. We subsequently demonstrate that Smad3 null MEFs are partially resistant to TGF-beta's antiproliferative effect, thus firmly establishing a role for Smad3 in TGF-beta-mediated growth inhibition. We next examined cells in which Smad3 is most highly expressed, specifically cells of immune origin. Although no specific developmental defect was detected in the immune system of the Smad3 null mice, a functional defect was observed in the ability of TGF-beta to inhibit the proliferation of splenocytes activated by specific stimuli. In addition, primary splenocytes display defects in TGF-beta-mediated repression of cytokine production. These data, taken together, establish a role for Smad3 in mediating the antiproliferative effects of TGF-beta and implicate Smad3 as a potential effector for TGF-beta in modulating immune system function.

Authors
Datto, MB; Frederick, JP; Pan, L; Borton, AJ; Zhuang, Y; Wang, XF
MLA Citation
Datto, MB, Frederick, JP, Pan, L, Borton, AJ, Zhuang, Y, and Wang, XF. "Targeted disruption of Smad3 reveals an essential role in transforming growth factor beta-mediated signal transduction." Mol Cell Biol 19.4 (April 1999): 2495-2504.
PMID
10082515
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
19
Issue
4
Publish Date
1999
Start Page
2495
End Page
2504

Smad3-Smad4 and AP-1 complexes synergize in transcriptional activation of the c-Jun promoter by transforming growth factor beta.

Transcriptional regulation by transforming growth factor beta (TGF-beta) is a complex process which is likely to involve cross talk between different DNA responsive elements and transcription factors to achieve maximal promoter activation and specificity. Here, we describe a concurrent requirement for two discrete responsive elements in the regulation of the c-Jun promoter, one a binding site for a Smad3-Smad4 complex and the other an AP-1 binding site. The two elements are located 120 bp apart in the proximal c-Jun promoter, and each was able to independently bind its corresponding transcription factor complex. The effects of independently mutating each of these elements were nonadditive; disruption of either sequence resulted in complete or severe reductions in TGF-beta responsiveness. This simultaneous requirement for two distinct and independent DNA binding elements suggests that Smad and AP-1 complexes function synergistically to mediate TGF-beta-induced transcriptional activation of the c-Jun promoter.

Authors
Wong, C; Rougier-Chapman, EM; Frederick, JP; Datto, MB; Liberati, NT; Li, JM; Wang, XF
MLA Citation
Wong, C, Rougier-Chapman, EM, Frederick, JP, Datto, MB, Liberati, NT, Li, JM, and Wang, XF. "Smad3-Smad4 and AP-1 complexes synergize in transcriptional activation of the c-Jun promoter by transforming growth factor beta." Mol Cell Biol 19.3 (March 1999): 1821-1830.
PMID
10022869
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
19
Issue
3
Publish Date
1999
Start Page
1821
End Page
1830

TGF-beta-induced phosphorylation of Smad3 regulates its interaction with coactivator p300/CREB-binding protein.

Smads are intermediate effector proteins that transduce the TGF-beta signal from the plasma membrane to the nucleus, where they participate in transactivation of downstream target genes. We have shown previously that coactivators p300/CREB-binding protein are involved in TGF-beta-mediated transactivation of two Cdk inhibitor genes, p21 and p15. Here we examined the possibility that Smads function to regulate transcription by directly interacting with p300/CREB-binding protein. We show that Smad3 can interact with a C-terminal fragment of p300 in a temporal and phosphorylation-dependent manner. TGF-beta-mediated phosphorylation of Smad3 potentiates the association between Smad3 and p300, likely because of an induced conformational change that removes the autoinhibitory interaction between the N- and C-terminal domains of Smad3. Consistent with a role for p300 in the transcription regulation of multiple genes, overexpression of a Smad3 C-terminal fragment causes a general squelching effect on multiple TGF-beta-responsive reporter constructs. The adenoviral oncoprotein E1A can partially block Smad-dependent transcriptional activation by directly competing for binding to p300. Taken together, these findings define a new role for phosphorylation of Smad3: in addition to facilitating complex formation with Smad4 and promoting nuclear translocation, the phosphorylation-induced conformational change of Smad3 modulates its interaction with coactivators, leading to transcriptional regulation.

Authors
Shen, X; Hu, PP; Liberati, NT; Datto, MB; Frederick, JP; Wang, XF
MLA Citation
Shen, X, Hu, PP, Liberati, NT, Datto, MB, Frederick, JP, and Wang, XF. "TGF-beta-induced phosphorylation of Smad3 regulates its interaction with coactivator p300/CREB-binding protein." Mol Biol Cell 9.12 (December 1998): 3309-3319.
PMID
9843571
Source
pubmed
Published In
Molecular Biology of the Cell
Volume
9
Issue
12
Publish Date
1998
Start Page
3309
End Page
3319

Molecular mechanisms of transforming growth factor-beta signaling.

Authors
Hu, PP; Datto, MB; Wang, XF
MLA Citation
Hu, PP, Datto, MB, and Wang, XF. "Molecular mechanisms of transforming growth factor-beta signaling." Endocr Rev 19.3 (June 1998): 349-363. (Review)
PMID
9626558
Source
pubmed
Published In
Endocrine reviews
Volume
19
Issue
3
Publish Date
1998
Start Page
349
End Page
363
DOI
10.1210/edrv.19.3.0333

Sp1, but not Sp3, functions to mediate promoter activation by TGF-beta through canonical Sp1 binding sites.

Transforming growth factor beta (TGF-beta) causes growth arrest at the G1 phase of the cell cycle in most cell types. Both the cyclin dependent kinase inhibitor p15(INK4B) and p21(Cip1/WAF1) genes have been found to be induced by TGF-beta in human keratinocyte HaCaT cells. Analyses of the human p15 and p21 promoters have led to the identification of GC-rich sequences capable of binding to Sp1 transcription factors as necessary elements for the TGF-beta induction of both promoters. We report here that canonical Sp1 binding sites derived from the SV40 21 bp repeat could also support promoter induction by TGF-beta when placed upstream of a minimal luciferase reporter construct containing only the TATA and Inr elements. Gel retardation assays identified Sp1, Sp3 and DeltaSp3 as major factors binding to the canonical Sp1 sites in HaCaT cells and that TGF-beta treatment did not change their binding activities over a 24 h period. More importantly, GAL4-Sp1, but not GAL4-Sp3, chimeric protein supported TGF-beta mediated gene induction from a luciferase reporter construct driven by five GAL4 DNA binding sites. Our results suggest that Sp1 binding site can function as a distinct TGF-beta responsive element for TGF-beta mediated promoter expression and Sp1 per se can mediate this response.

Authors
Li, JM; Datto, MB; Shen, X; Hu, PP; Yu, Y; Wang, XF
MLA Citation
Li, JM, Datto, MB, Shen, X, Hu, PP, Yu, Y, and Wang, XF. "Sp1, but not Sp3, functions to mediate promoter activation by TGF-beta through canonical Sp1 binding sites." Nucleic Acids Res 26.10 (May 15, 1998): 2449-2456.
PMID
9580699
Source
pubmed
Published In
Nucleic Acids Research
Volume
26
Issue
10
Publish Date
1998
Start Page
2449
End Page
2456

TGF-β-induced phosphorylation of Smad3 regulates its interaction with coactivator p300/CREB-binding protein

Smads are intermediate effector proteins that transduce the TGF-β signal from the plasma membrane to the nucleus, where they participate in transactivation of downstream target genes. We have shown previously that coactivators p300/CREB-binding protein are involved in TGF-β-mediated transactivation of two Cdk inhibitor genes, p21 and p15. Here we examined the possibility that Smads function to regulate transcription by directly interacting with p300/CREB-binding protein. We show that Smad3 can interact with a C-terminal fragment of p300 in a temporal and phosphorylation- dependent manner. TGF-β-mediated phosphorylation of Smad3 potentiates the association between Smad3 and p300, likely because of an induced conformational change that removes the autoinhibitory interaction between the N- and C-terminal domains of Smad3. Consistent with a role for p300 in the transcription regulation of multiple genes, overexpression of a Smad3 C- terminal fragment causes a general squelching effect on multiple TGF-β- responsive reporter constructs. The adenoviral oncoprotein E1A can partially block Smad-dependent transcriptional activation by directly competing for binding to p300. Taken together, these findings define a new role for phosphorylation of Smad3: in addition to facilitating complex formation with Smad4 and promoting nuclear translocation, the phosphorylation-induced conformational change of Smad3 modulates its interaction with coactivators, leading to transcriptional regulation.

Authors
Shen, X; Hu, PP-C; Liberati, NT; Datto, MB; Frederick, JP; Wang, X-F
MLA Citation
Shen, X, Hu, PP-C, Liberati, NT, Datto, MB, Frederick, JP, and Wang, X-F. "TGF-β-induced phosphorylation of Smad3 regulates its interaction with coactivator p300/CREB-binding protein." Molecular Biology of the Cell 9.12 (1998): 3309-3319.
Source
scival
Published In
Molecular Biology of the Cell
Volume
9
Issue
12
Publish Date
1998
Start Page
3309
End Page
3319

Tumor suppressor Smad4 is a transforming growth factor beta-inducible DNA binding protein.

Members of the Smad family of proteins are thought to play important roles in transforming growth factor beta (TGF-beta)-mediated signal transduction. In response to TGF-beta, specific Smads become inducibly phosphorylated, form heteromers with Smad4, and undergo nuclear accumulation. In addition, overexpression of specific Smad combinations can mimic the transcriptional effect of TGF-beta on both the plasminogen activator inhibitor 1 (PAI-1) promoter and the reporter construct p3TP-Lux. Although these data suggest a role for Smads in regulating transcription, the precise nuclear function of these heteromeric Smad complexes remains largely unknown. Here we show that in Mv1Lu cells Smad3 and Smad4 form a TGF-beta-induced, phosphorylation-dependent, DNA binding complex that specifically recognizes a bipartite binding site within p3TP-Lux. Furthermore, we demonstrate that Smad4 itself is a DNA binding protein which recognizes the same sequence. Interestingly, mutations which eliminate the Smad DNA binding site do not interfere with either TGF-beta-dependent transcriptional activation or activation by Smad3/Smad4 cooverexpression. In contrast, mutation of adjacent AP1 sites within this context eliminates both TGF-beta-dependent transcriptional activation and activation in response to Smad3/Smad4 cooverexpression. Furthermore, concatemerized AP1 sites, in isolation, are activated by Smad3/Smad4 cooverexpression and, to a certain extent, by TGF-beta. Taken together, these data suggest that the Smad3/Smad4 complex has at least two separable nuclear functions: it forms a rapid, yet transient sequence-specific DNA binding complex, and it potentiates AP1-dependent transcriptional activation.

Authors
Yingling, JM; Datto, MB; Wong, C; Frederick, JP; Liberati, NT; Wang, XF
MLA Citation
Yingling, JM, Datto, MB, Wong, C, Frederick, JP, Liberati, NT, and Wang, XF. "Tumor suppressor Smad4 is a transforming growth factor beta-inducible DNA binding protein." Mol Cell Biol 17.12 (December 1997): 7019-7028.
PMID
9372933
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
17
Issue
12
Publish Date
1997
Start Page
7019
End Page
7028

The viral oncoprotein E1A blocks transforming growth factor beta-mediated induction of p21/WAF1/Cip1 and p15/INK4B.

The adenovirus early gene product E1A is a potent stimulator of cellular proliferation, which when overexpressed can overcome the growth-inhibitory effects of the polypeptide hormone transforming growth factor beta (TGF-beta). The ability of TGF-beta to arrest cell growth in G1 correlates with the transcriptional induction of the cyclin-dependent kinase inhibitors, p15/INK4B and p21/WAF1/Cip1; an inhibition of the G1 cyclin-Cdk complexes; and a maintenance of the retinoblastoma susceptibility gene product, Rb, in a hypophosphorylated state. The ability of E1A to overcome TGF-beta-mediated growth inhibition derives, in part, from its ability to sequester Rb and Rb family members. We report here that E1A also acts upstream of Rb by blocking the TGF-beta-mediated induction of p15 and p21. Consistent with these findings, E1A expression also blocks the ability of TGF-beta to inhibit Cdk2 kinase activity, as well as its ability to hold Rb in a hypophosphorylated state. The effect of E1A on the induction of p15 and p21 is independent of E1A's Rb binding activity. The E1A-mediated decrease in p15 levels is primarily the result of a block at the level of transcriptional activation by TGF-beta. This effect is dependent on E1A's ability to bind p300, one of E1A's target proteins. Thus, the ability of E1A to affect p15 and p21 expression represents an additional possible mechanism by which E1A can circumvent the negative regulation of cell cycle progression.

Authors
Datto, MB; Hu, PP; Kowalik, TF; Yingling, J; Wang, XF
MLA Citation
Datto, MB, Hu, PP, Kowalik, TF, Yingling, J, and Wang, XF. "The viral oncoprotein E1A blocks transforming growth factor beta-mediated induction of p21/WAF1/Cip1 and p15/INK4B." Mol Cell Biol 17.4 (April 1997): 2030-2037.
PMID
9121451
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
17
Issue
4
Publish Date
1997
Start Page
2030
End Page
2037

The TGFβ receptors and signaling pathways

Authors
Datto, MB; Bassing, CH; Wang, X-F
MLA Citation
Datto, MB, Bassing, CH, and Wang, X-F. "The TGFβ receptors and signaling pathways." Growth Factors and Cytokines in Health and Disease 1.C (1996): 395-432.
Source
scival
Published In
Growth Factors and Cytokines in Health and Disease
Volume
1
Issue
C
Publish Date
1996
Start Page
395
End Page
432
DOI
10.1016/S1874-5687(96)80017-6

Functional analysis of the transforming growth factor beta responsive elements in the WAF1/Cip1/p21 promoter.

The transforming growth factor beta s (TGF-beta s) are a group of multifunctional growth factors that inhibit cell cycle progression in many cell types. The TGF-beta-induced cell cycle arrest has been partially attributed to the regulatory effects of TGF-beta on both the levels and activities of the G1 cyclins and their cyclin-dependent kinase partners. The ability of TGF-beta to inhibit the activity of these kinase complexes derives in part from its regulatory effects on the cyclin-dependent kinase inhibitors, p21/WAF1/Cip1, p27Kip1, and p15. Upon treatment of cells with TGF-beta, these three inhibitors bind to and block the activities of specific cyclin-cyclin-dependent kinase complexes to cause cell cycle arrest. Little is known, however, on the mechanism through which TGF-beta activates these cyclin-dependent kinase inhibitors. In the case of p21, TGF-beta treatment leads to an increase in p21 mRNA. This increase in p21 mRNA is partly due to transcriptional activation of the p21 promoter by TGF-beta. To further define the signaling pathways through which TGF-beta induces p21, we have performed a detailed functional analysis on the p21 promoter. Through both deletion and mutation analysis of the p21 promoter, we have defined a 10-base pair sequence that is required for the activation of the p21 promoter by TGF-beta. In addition, this sequence is sufficient to drive TGF-beta-mediated transcription from a previously nonresponsive promoter. Preliminary gel shift assays demonstrate that this TGF-beta responsive element binds specifically to several proteins in vitro. Two of these proteins are the transcription factors Sp-1 and Sp-3. These studies represent the initial steps toward defining the signaling pathways involved in TGF-beta-mediated transcriptional activation of p21.

Authors
Datto, MB; Yu, Y; Wang, XF
MLA Citation
Datto, MB, Yu, Y, and Wang, XF. "Functional analysis of the transforming growth factor beta responsive elements in the WAF1/Cip1/p21 promoter." J Biol Chem 270.48 (December 1, 1995): 28623-28628.
PMID
7499379
Source
pubmed
Published In
The Journal of biological chemistry
Volume
270
Issue
48
Publish Date
1995
Start Page
28623
End Page
28628

Transforming growth factor beta induces the cyclin-dependent kinase inhibitor p21 through a p53-independent mechanism.

The transforming growth factor beta s (TGF-beta s) are a group of multifunctional growth factors which inhibit cell cycle progression in many cell types. The TGF-beta-induced cell cycle arrest has been partially attributed to the regulatory effects of TGF-beta on both the levels and the activities of the G1 cyclins and their kinase partners. The activities of these kinases are negatively regulated by a number of small proteins, p21 (WAF1, Cip1), p27Kip1, p16, and p15INK4B, that physically associate with cyclins, cyclin-dependent kinases, or cyclin-Cdk complexes. p21 has been previously shown to be transcriptionally induced by DNA damage through p53 as a mediator. We demonstrate that TGF-beta also causes a rapid transcriptional induction of p21, suggesting that p21 can respond to both intracellular and extracellular signals for cell cycle arrest. In contrast to DNA damage, however, induction of p21 by TGF-beta is not dependent on wild-type p53. The cell line studied in these experiments, HaCaT, contains two mutant alleles of p53, which are unable to activate transcription from the p21 promoter when overexpressed. In addition, TGF-beta and p53 act through distinct elements in the p21 promoter. Taken together, these findings suggest that TGF-beta can induce p21 through a p53-independent pathway. Previous findings have implicated p27Kip1 and p15INK2B as effectors mediating the TGF-beta growth inhibitory effect. These results demonstrate that a single extracellular antiproliferative signal, TGF-beta, can act through multiple signaling pathways to elicit a growth arrest response.

Authors
Datto, MB; Li, Y; Panus, JF; Howe, DJ; Xiong, Y; Wang, XF
MLA Citation
Datto, MB, Li, Y, Panus, JF, Howe, DJ, Xiong, Y, and Wang, XF. "Transforming growth factor beta induces the cyclin-dependent kinase inhibitor p21 through a p53-independent mechanism." Proc Natl Acad Sci U S A 92.12 (June 6, 1995): 5545-5549.
PMID
7777546
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
92
Issue
12
Publish Date
1995
Start Page
5545
End Page
5549

Expression of the E2F1 transcription factor overcomes type beta transforming growth factor-mediated growth suppression.

Inhibition of cell growth by type beta transforming growth factor (TGF-beta) occurs in mid-G1 and is associated with decreased G1 cyclin-dependent kinase activity and maintenance of the retinoblastoma tumor suppressor protein Rb in an underphosphorylated, growth-suppressive state. A variety of recent experiments suggest that a functional target of Rb is the E2F transcription factor. In addition, the growth-suppressive effects of TGF-beta can be overcome by expression of viral oncogene products that dissociate E2F from Rb and Rb-related polypeptides. These results suggest the possibility that control of E2F may be a downstream event of TGF-beta action. Consistent with that possibility is the observation that E2F1 RNA levels are drastically reduced in TGF-beta-treated cells. We have also used a recombinant adenovirus containing the human E2F1 gene to overexpress the E2F1 product in mink lung epithelial cells that were growth arrested with TGF-beta. We find that overexpression of E2F1 can overcome the TGF-beta-mediated effect as measured by the activation of cellular DNA synthesis. These results suggest that a likely downstream target for the cyclin-dependent kinases, which are controlled by TGF-beta, is the activation of E2F.

Authors
Schwarz, JK; Bassing, CH; Kovesdi, I; Datto, MB; Blazing, M; George, S; Wang, XF; Nevins, JR
MLA Citation
Schwarz, JK, Bassing, CH, Kovesdi, I, Datto, MB, Blazing, M, George, S, Wang, XF, and Nevins, JR. "Expression of the E2F1 transcription factor overcomes type beta transforming growth factor-mediated growth suppression." Proc Natl Acad Sci U S A 92.2 (January 17, 1995): 483-487.
PMID
7831315
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
92
Issue
2
Publish Date
1995
Start Page
483
End Page
487
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