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Zhang, Yunyan

Overview:

Epidermis of the skin undergoes lifelong self-renewal through a tight balance of cell growth, differentiation and programmed cell death. Deregulation of this balance is manifested in many diseases, including autoimmune diseases, psoriasis and cancer. Our lab is focused on addressing the following questions: 1) how are AP-1 family transcription factors involved in regulating human epithelial homeostasis and neoplasia? 2) how do NF-κB and JNK signaling pathways cross-talk to promote human squamous cell carcinoma (SCC)? and 3) What are the molecular mechanisms mediating the function of CYLD tumor suppressor in human SCC and melanoma?

I. Genetic regulation of epithelial cell proliferation and differentiation
NF-κB and AP-1 family gene regulatory proteins have been indicated in a wide range of cellular processes. Using both murine genetic and regenerated human skin tissue models, we have recently demonstrated that NF-κB and AP-1 regulate epidermal cell growth in an apposing fashion with the former inducing cell growth arrest and the later promoting cell proliferation. On the other hand, recent reports have revealed functional diversity for different AP-1 subunits. In particular, JunB and c-Jun AP-1 subunits have been implicated in a mouse model of psoriasis, while their role in human psoriasis remains controversial. Our goal is to use human skin tissue models to determine how JunB and c-Jun regulate epidermal cell proliferation and differentiation, and to explore their relevance to human skin diseases, including skin cancer and psoriasis.

II. Signaling networks in human SCC
Ras activation has emerged as a hallmark feature of a majority of human epidermal cancers, including SCC, the second most common malignancy in the United States. In addition to Ras induction, IKK/NF-κB loss-of-function and JNK/AP-1 gain-of-function have been established in spontaneous human SCC. In order to study interactions between these dominant signaling cascades, we use multiplex serial gene transfer (MSGT) through which alterations in multiple signaling networks can be rapidly made in normal cells. Combined with tissue engineering, MSGT permits the molecular reconstruction of events sufficient to turn normal human tissues into cancer. These new genetic approaches have led to the finding that either NF-κB blockade or JNK induction is sufficient to act in conjunction with Ras activation to transform normal human epidermal cells directly into invasive neoplasia. Our current ongoing efforts are directed at 1) exploring the mechanisms governing the JNK signaling pathway in epidermal carcinogenesis; and 2) exploring therapeutic values of blocking JNK-AP1 signaling for human SCC.

III. CYLD in human SCC and melanoma
CYLD, a deubiquitinase, is initially identified as a tumor suppressor due to its autosomal dominant genetic linkage to skin appendage tumors collectively named Brook-Speigler Syndrome (BSS). The role of CYLD has been recently expanded to many other cancers, including melanoma, colon, liver, lung and renal and hematopoietic cancers. However, the underlying molecular mechanisms of CYLD in tumorigenesis are still unclear. Using transgenic mouse model, we have demonstrated that K14-driven epidermal specific expression of a catalytically deficient CYLD mutant (CYLDm) leads to increased sensitivity to chemically induced skin carcinogenesis. We have also found that CYLD is downregulated in human SCC and melanoma. Most importantly, restoring CYLD expression inhibits tumorigenesis of both SCC and melanoma. Currently, our efforts are directed at understanding 1) mechanisms mediating the CYLD expression, 2) the molecular mechanisms governing CYLD function in nonmelanoma and melanoma skin cancer.

Positions:

Associate Professor in Dermatology

Dermatology
School of Medicine

Assistant Professor in Pathology

Pathology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

Ph.D. 1998

Ph.D. — University of Florida

Grants:

Spindle Orientation in Skin Development and Homeostasis

Administered By
Dermatology
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
September 01, 2015
End Date
August 31, 2020

Pilot Study Evaluating the Efficacy of a Topical PDE-4 Inhibitor for Morphea

Administered By
Dermatology
AwardedBy
Pfizer, Inc.
Role
Co-Principal Investigator
Start Date
August 14, 2017
End Date
December 31, 2019

THE ROLE OF MALT1 IN MELANOMA GROWTH AND METASTASIS

Administered By
Dermatology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 01, 2015
End Date
June 30, 2018

Improving Melanoma Diagnosis with Pump-Probe Optical Imaging

Administered By
Chemistry
AwardedBy
National Institutes of Health
Role
Co Investigator
Start Date
January 02, 2013
End Date
December 31, 2017

JUN PROTEINS IN EPIDERMAL HOMEOSTASIS AND NEOPLASIA

Administered By
Dermatology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
August 09, 2010
End Date
May 31, 2015

Imaging Nonlinear Absorption of Biomarkers for Improved Detection of Melanoma

Administered By
Chemistry
AwardedBy
National Institutes of Health
Role
Investigator
Start Date
September 30, 2009
End Date
March 29, 2012

Role of c-Jun N-terminal kinases in human epidermal homeostasis

Administered By
Dermatology
AwardedBy
National Institutes of Health
Role
Sponsor
Start Date
July 23, 2009
End Date
September 15, 2010

CYLD Regulation of Epidermal Growth and Neoplasia

Administered By
Dermatology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
August 01, 2007
End Date
July 31, 2010

NF-kB, CDK4 and JNK in Epidermal Growth Regulation

Administered By
Dermatology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
April 05, 2005
End Date
March 31, 2010
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Publications:

MALT1 promotes melanoma progression through JNK/c-Jun signaling.

Mucosa-associated lymphoma antigen 1 (MALT1) is a lymphoma oncogene that regulates signal transduction as a paracaspase and an adaptor protein. Yet, the role of MALT1 in other solid cancers such as melanoma is not well-understood. Here, we demonstrate that MALT1 is overexpressed in malignant melanoma cells, and predicts a poor disease-free survival. MALT1 inhibition via shRNA-mediated gene silencing or pharmacologically with MI-2 compound markedly reduced cell growth and migration of A2058 and A375 melanoma cell lines in vitro. Subcutaneous tumor growth analysis revealed that MALT1 gene silencing significantly reduced tumor growth and metastasis to the lung. Consistently, the subcutaneous tumors with MALT1 loss had increased cell apoptosis and decreased proliferation. In addition, these tumors showed signs of mesenchymal-epithelial transition as indicated by the upregulation of E-cadherin and downregulation of N-cadherin and β1-intergrin. Further molecular analysis revealed that MALT1 is required for c-Jun and nuclear factor-κB (NF-κB) activation by tumor necrosis factor-α. Forced expression of the c-Jun upstream activator MKK7 reversed the cell growth and migration defects caused by MALT1 loss. In contrast, NF-κB activation via expression of p65ER, a fusion protein containing NF-κB p65 and the tamoxifen-responsive mutant estrogen receptor, induced minimal effects on cell proliferation, but diminished cell death induced by MALT1 loss and TRAIL treatment. Together, these findings demonstrate that MALT1 promotes melanoma cell proliferation and motility through JNK/c-Jun, and enhances melanoma cell survival through NF-κB, underscoring MALT1 as a potential therapeutic target and biomarker for malignant melanoma.

Authors
Wang, Y; Zhang, G; Jin, J; Degan, S; Tameze, Y; Zhang, JY
MLA Citation
Wang, Y, Zhang, G, Jin, J, Degan, S, Tameze, Y, and Zhang, JY. "MALT1 promotes melanoma progression through JNK/c-Jun signaling." Oncogenesis 6.7 (July 31, 2017): e365-.
Website
http://hdl.handle.net/10161/15170
PMID
28759024
Source
epmc
Published In
Oncogenesis
Volume
6
Issue
7
Publish Date
2017
Start Page
e365
DOI
10.1038/oncsis.2017.68

Animal models of skin disordersThe role of the c-Jun N-terminal Kinase signaling pathway in skin cancer

Animal Models for the Study of Human Disease, Second Edition, provides needed information on model sharing, animal alternatives, animal ethics and access to databanks of models, bringing together common descriptions of models for busy ...

Authors
Zhang, JY; Zhang, JY; Selim, MA
MLA Citation
Zhang, JY, Zhang, JY, and Selim, MA. "Animal models of skin disordersThe role of the c-Jun N-terminal Kinase signaling pathway in skin cancer." Animal Models for the Study of Human Disease. Ed. PM Conn. Academic Press, June 20, 2017. 357-371. (Chapter)
Website
http://hdl.handle.net/10161/13287
Source
manual
Publish Date
2017
Start Page
357
End Page
371

Printing amphotericin B on microneedles using matrix assisted pulsed laser evaporation

Authors
Sachan, R; Jaipan, P; Zhang, JY; Degan, S; Erdmann, D; Tedesco, J; Vanderwal, L; Stafslien, SJ; Negut, I; Visan, A; Dorcioman, G; Socol, G; Cristescu, R; Chrisey, DB; Narayan, RJ
MLA Citation
Sachan, R, Jaipan, P, Zhang, JY, Degan, S, Erdmann, D, Tedesco, J, Vanderwal, L, Stafslien, SJ, Negut, I, Visan, A, Dorcioman, G, Socol, G, Cristescu, R, Chrisey, DB, and Narayan, RJ. "Printing amphotericin B on microneedles using matrix assisted pulsed laser evaporation." International Journal of Bioprinting 3.2 (June 12, 2017).
Source
crossref
Volume
3
Issue
2
Publish Date
2017
DOI
10.18063/IJB.2017.02.004

TRPV4 Moves toward Center-Fold in Rosacea Pathogenesis.

Mascarenhas et al. report that TRPV4 expression is upregulated in mast cells in response to the proteolytic cathelicidin fragment LL37 in a murine rosacea model and that TRPV4 loss of function attenuates mast cell degranulation. These findings render TRPV4 a translational-medical target in rosacea. However, signaling mechanisms causing increased expression of TRPV4 await elucidation. Moreover, we ask whether TRPV4-mediated Ca++-influx evokes mast cell degranulation.

Authors
Chen, Y; Moore, CD; Zhang, JY; Hall, RP; MacLeod, AS; Liedtke, W
MLA Citation
Chen, Y, Moore, CD, Zhang, JY, Hall, RP, MacLeod, AS, and Liedtke, W. "TRPV4 Moves toward Center-Fold in Rosacea Pathogenesis." The Journal of Investigative Dermatology 137.4 (April 2017): 801-804.
PMID
28340683
Source
epmc
Published In
Journal of Investigative Dermatology
Volume
137
Issue
4
Publish Date
2017
Start Page
801
End Page
804
DOI
10.1016/j.jid.2016.12.013

KIND1 Loss Sensitizes Keratinocytes to UV-Induced Inflammatory Response and DNA Damage.

Loss of function of KIND1, a cytoskeletal protein involved in β1-integrin function, causes Kindler syndrome, a genetic disease characterized by skin fragility, photosensitivity, and increased risk of squamous cell carcinoma. Dysregulation of β1-integrin underlies Kindler syndrome skin fragility. However, the mechanisms underlying squamous cell carcinoma susceptibility are unclear. Here, we demonstrate that gene silencing of KIND1 decreased keratinocyte proliferation and increased apoptosis in vitro and in skin grafts regenerated on mice, which was correlated with reduced cyclinB1. In addition, KIND1 loss sensitized keratinocytes to cytokine and UV-induced NF-κB and c-Jun N-terminal kinase activation and upregulation of CXCL10 and tumor necrosis factor-α. Moreover, KIND1 loss impaired DNA repair, as indicated by the increased detection of γH2AX and cyclobutane pyrimidine dimers 24 hours after UVB radiation. Genetic or pharmacological c-Jun N-terminal kinase inhibition and NF-κB inhibition markedly reduced cyclobutane pyrimidine dimers-positive cells. Further, we show that KIND1 was regulated by JunB at the transcriptional level and, like JunB, it was downregulated in human squamous cell carcinoma cells. Together, these results indicate that KIND1 is important not only for keratinocyte proliferation but also for the suppression of UV-induced inflammation and DNA damage. These latter findings support a tumor suppressor function for KIND1, and identify c-Jun N-terminal kinase and NF-κB as potential therapeutic targets for prevention of squamous cell carcinoma in patients with Kindler syndrome.

Authors
Zhang, X; Luo, S; Wu, J; Zhang, L; Wang, W-H; Degan, S; Erdmann, D; Hall, R; Zhang, JY
MLA Citation
Zhang, X, Luo, S, Wu, J, Zhang, L, Wang, W-H, Degan, S, Erdmann, D, Hall, R, and Zhang, JY. "KIND1 Loss Sensitizes Keratinocytes to UV-Induced Inflammatory Response and DNA Damage." The Journal of investigative dermatology 137.2 (February 2017): 475-483.
PMID
27725201
Source
epmc
Published In
Journal of Investigative Dermatology
Volume
137
Issue
2
Publish Date
2017
Start Page
475
End Page
483
DOI
10.1016/j.jid.2016.09.023

Epidermal CYLD inactivation sensitizes mice to the development of sebaceous and basaloid skin tumors.

The deubiquitinase-encoding gene Cyld displays a dominant genetic linkage to a wide spectrum of skin-appendage tumors, which could be collectively designated as CYLD mutant-syndrome (CYLD(m)-syndrome). Despite recent advances, little is understood about the molecular mechanisms responsible for this painful and difficult-to-treat skin disease. Here, we generated a conditional mouse model with epidermis-targeted expression of a catalytically deficient CYLD(m) through K14-Cre-mediated deletion of exon 9 (hereafter refer to Cyld(EΔ9/Δ9) ). Cyld(EΔ9/Δ9) mice were born alive but developed hair and sebaceous gland abnormalities and dental defects at 100% and 60% penetrance, respectively. Upon topical challenge with DMBA/TPA, these animals primarily developed sebaceous and basaloid tumors resembling human CYLD(m)-syndrome as opposed to papilloma, which is most commonly induced in WT mice by this treatment. Molecular analysis revealed that TRAF6-K63-Ubiquitination (K63-Ub), c-Myc-K63-Ub, and phospho-c-Myc (S62) were markedly elevated in Cyld(EΔ9/Δ9) skin. Topical treatment with a pharmacological c-Myc inhibitor induced sebaceous and basal cell apoptosis in Cyld(EΔ9/Δ9) skin. Consistently, c-Myc activation was readily detected in human cylindroma and sebaceous adenoma. Taken together, our findings demonstrate that Cyld(EΔ9/Δ9) mice represent a disease-relevant animal model and identify TRAF6 and c-Myc as potential therapeutic targets for CYLD(m)-syndrome.

Authors
Jin, YJ; Wang, S; Cho, J; Selim, MA; Wright, T; Mosialos, G; Zhang, JY
MLA Citation
Jin, YJ, Wang, S, Cho, J, Selim, MA, Wright, T, Mosialos, G, and Zhang, JY. "Epidermal CYLD inactivation sensitizes mice to the development of sebaceous and basaloid skin tumors." JCI insight 1.11 (July 2016).
PMID
27478875
Source
epmc
Published In
JCI insight
Volume
1
Issue
11
Publish Date
2016

FRA1 promotes squamous cell carcinoma growth and metastasis through distinct AKT and c-Jun dependent mechanisms.

FRA1 (Fos-like antigen 1) is highly expressed in many epithelial cancers including squamous cell carcinoma of the skin (cSCC) and head and neck (HNSCC). However, the functional importance and the mechanisms mediating FRA1 function in these cancers are not fully understood. Here, we demonstrate that FRA1 gene silencing in HNSCC and cSCC cells resulted in two consequences - impaired cell proliferation and migration. FRA1 regulation of cell growth was distinct from that of c-Jun, a prominent Jun group AP-1 factor. While c-Jun was required for the expression of the G1/S phase cell cycle promoter CDK4, FRA1 was essential for AKT activation and AKT-dependent expression of CyclinB1, a molecule required for G2-M progression. Exogenous expression of a constitutively active form of AKT rescued cancer cell growth defect caused by FRA1-loss. Additionally, FRA1 knockdown markedly slowed cell adhesion and migration, and conversely expression of an active FRA1 mutant (FRA1DD) expedited these processes in a JNK/c-Jun-dependent manner. Through protein and ChIP-PCR analyses, we identified KIND1, a cytoskeletal regulator of the cell adhesion molecule β1-integrin, as a novel FRA1 transcriptional target. Restoring KIND1 expression rescued migratory defects induced by FRA1 loss. In agreement with these in vitro data, HNSCC cells with FRA1 loss displayed markedly reduced rates of subcutaneous tumor growth and pulmonary metastasis. Together, these results indicate that FRA1 promotes cancer growth through AKT, and enhances cancer cell migration through JNK/c-Jun, pinpointing FRA1 as a key integrator of JNK and AKT signaling pathways and a potential therapeutic target for cSCC and HNSCC.

Authors
Zhang, X; Wu, J; Luo, S; Lechler, T; Zhang, JY
MLA Citation
Zhang, X, Wu, J, Luo, S, Lechler, T, and Zhang, JY. "FRA1 promotes squamous cell carcinoma growth and metastasis through distinct AKT and c-Jun dependent mechanisms." Oncotarget 7.23 (June 2016): 34371-34383.
Website
http://hdl.handle.net/10161/15165
PMID
27144339
Source
epmc
Published In
Oncotarget
Volume
7
Issue
23
Publish Date
2016
Start Page
34371
End Page
34383
DOI
10.18632/oncotarget.9110

Transient Receptor Potential Vanilloid 4 Ion Channel Functions as a Pruriceptor in Epidermal Keratinocytes to Evoke Histaminergic Itch.

TRPV4 ion channels function in epidermal keratinocytes and in innervating sensory neurons; however, the contribution of the channel in either cell to neurosensory function remains to be elucidated. We recently reported TRPV4 as a critical component of the keratinocyte machinery that responds to ultraviolet B (UVB) and functions critically to convert the keratinocyte into a pain-generator cell after excess UVB exposure. One key mechanism in keratinocytes was increased expression and secretion of endothelin-1, which is also a known pruritogen. Here we address the question of whether TRPV4 in skin keratinocytes functions in itch, as a particular form of "forefront" signaling in non-neural cells. Our results support this novel concept based on attenuated scratching behavior in response to histaminergic (histamine, compound 48/80, endothelin-1), not non-histaminergic (chloroquine) pruritogens in Trpv4 keratinocyte-specific and inducible knock-out mice. We demonstrate that keratinocytes rely on TRPV4 for calcium influx in response to histaminergic pruritogens. TRPV4 activation in keratinocytes evokes phosphorylation of mitogen-activated protein kinase, ERK, for histaminergic pruritogens. This finding is relevant because we observed robust anti-pruritic effects with topical applications of selective inhibitors for TRPV4 and also for MEK, the kinase upstream of ERK, suggesting that calcium influx via TRPV4 in keratinocytes leads to ERK-phosphorylation, which in turn rapidly converts the keratinocyte into an organismal itch-generator cell. In support of this concept we found that scratching behavior, evoked by direct intradermal activation of TRPV4, was critically dependent on TRPV4 expression in keratinocytes. Thus, TRPV4 functions as a pruriceptor-TRP in skin keratinocytes in histaminergic itch, a novel basic concept with translational-medical relevance.

Authors
Chen, Y; Fang, Q; Wang, Z; Zhang, JY; MacLeod, AS; Hall, RP; Liedtke, WB
MLA Citation
Chen, Y, Fang, Q, Wang, Z, Zhang, JY, MacLeod, AS, Hall, RP, and Liedtke, WB. "Transient Receptor Potential Vanilloid 4 Ion Channel Functions as a Pruriceptor in Epidermal Keratinocytes to Evoke Histaminergic Itch." The Journal of biological chemistry 291.19 (May 2016): 10252-10262.
Website
http://hdl.handle.net/10161/12969
PMID
26961876
Source
epmc
Published In
The Journal of biological chemistry
Volume
291
Issue
19
Publish Date
2016
Start Page
10252
End Page
10262
DOI
10.1074/jbc.m116.716464

Use of Drawing Lithography-Fabricated Polyglycolic Acid Microneedles for Transdermal Delivery of Itraconazole to a Human Basal Cell Carcinoma Model Regenerated on Mice

Authors
Zhang, J; Wang, Y; Jin, JY; Degan, S; Hall, RP; Boehm, RD; Jaipan, P; Narayan, RJ
MLA Citation
Zhang, J, Wang, Y, Jin, JY, Degan, S, Hall, RP, Boehm, RD, Jaipan, P, and Narayan, RJ. "Use of Drawing Lithography-Fabricated Polyglycolic Acid Microneedles for Transdermal Delivery of Itraconazole to a Human Basal Cell Carcinoma Model Regenerated on Mice." JOM 68.4 (April 2016): 1128-1133.
Source
crossref
Published In
JOM
Volume
68
Issue
4
Publish Date
2016
Start Page
1128
End Page
1133
DOI
10.1007/s11837-016-1841-1

Comparing in vivo pump-probe and multiphoton fluorescence microscopy of melanoma and pigmented lesions.

We demonstrate a multimodal approach that combines a pump-probe with confocal reflectance and multiphoton autofluorescence microscopy. Pump-probe microscopy has been proven to be of great value in analyzing thin tissue sections of pigmented lesions, as it produces molecular contrast which is inaccessible by other means. However, the higher optical intensity required to overcome scattering in thick tissue leads to higher-order nonlinearities in the optical response of melanin (e.g., two-photon pump and one-photon probe) that present additional challenges for interpreting the data. We show that analysis of pigment composition in vivo must carefully account for signal terms that are nonlinear with respect to the pump and probe intensities. We find that pump-probe imaging gives useful contrast for pigmented structures over a large range of spatial scales (100 μm to 1 cm), making it a potentially useful tool for tracking the progression of pigmented lesions without the need to introduce exogenous contrast agents.

Authors
Wilson, JW; Degan, S; Gainey, CS; Mitropoulos, T; Simpson, MJ; Zhang, JY; Warren, WS
MLA Citation
Wilson, JW, Degan, S, Gainey, CS, Mitropoulos, T, Simpson, MJ, Zhang, JY, and Warren, WS. "Comparing in vivo pump-probe and multiphoton fluorescence microscopy of melanoma and pigmented lesions." Journal of Biomedical Optics 20.5 (May 2015): 051012-.
PMID
25415567
Source
epmc
Published In
Journal of Biomedical Optics
Volume
20
Issue
5
Publish Date
2015
Start Page
051012
DOI
10.1117/1.jbo.20.5.051012

RNA-Seq and ChIP-Seq reveal SQSTM1/p62 as a key mediator of JunB suppression of NF-κB-dependent inflammation.

Mice with epidermal deletion of JunB transcription factor displayed a psoriasis-like inflammation. The relevance of these findings to humans and the mechanisms mediating JunB function are not fully understood. Here we demonstrate that impaired JunB function via gene silencing or overexpression of a dominant negative mutant increased human keratinocyte cell proliferation but decreased cell barrier function. RNA-seq revealed over 500 genes affected by JunB loss of function, which included the upregulation of an array of proinflammatory molecules relevant to psoriasis. Among these were tumor necrosis factor α (TNFα), CCL2, CXCL10, IL6R, and SQSTM1, an adaptor protein involved in nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation. Chromatin immunoprecipitation (ChIP)-Seq and gene reporter analyses showed that JunB directly suppressed SQSTM1 by binding to a consensus AP-1 cis element located around 2 kb upstream of SQSTM1-transcription start site. Similar to JunB loss of function, SQSTM1-overexpression induced TNFα, CCL2, and CXCL10. Conversely, NF-κB inhibition genetically with a mutant IκBα or pharmacologically with pyrrolidine dithiocarbamate (PDTC) prevented cytokine, but not IL6R, induction by JunB deficiency. Taken together, our findings indicate that JunB controls epidermal growth, barrier formation, and proinflammatory responses through direct and indirect mechanisms, pinpointing SQSTM1 as a key mediator of JunB suppression of NF-κB-dependent inflammation.

Authors
Zhang, X; Jin, JY; Wu, J; Qin, X; Streilein, R; Hall, RP; Zhang, JY
MLA Citation
Zhang, X, Jin, JY, Wu, J, Qin, X, Streilein, R, Hall, RP, and Zhang, JY. "RNA-Seq and ChIP-Seq reveal SQSTM1/p62 as a key mediator of JunB suppression of NF-κB-dependent inflammation." The Journal of investigative dermatology 135.4 (April 2015): 1016-1024.
Website
http://hdl.handle.net/10161/15166
PMID
25501661
Source
epmc
Published In
Journal of Investigative Dermatology
Volume
135
Issue
4
Publish Date
2015
Start Page
1016
End Page
1024
DOI
10.1038/jid.2014.519

In vivo pump-probe microscopy of melanoma: Characterizing shifts in excited state photodynamics with respect to invasiveness

© 2015 SPIE. Pump-probe microscopy is a multiphoton technique that generates molecular contrast from absorptive pigments, such as melanin. It holds the potential to be used as a non-invasive screening tool to discern whether a given early-stage melanoma has acquired the capacity for metastasis. Here, we examined lesions in a Braf(V600E)-driven model of melanoma to assess whether loss of the tumor suppressor gene Pten in a is accompanied by a shift in pigment expression, as measured in vivo by pump-probe microscopy. The data were analyzed to determine differences in the excited-state lifetime of melanins expressed in Pten-competent and Pten-loss pigmented lesions. Loss of the tumor suppressor Pten was found to be accompanied by a statistically significant decrease in pixel-average excited state lifetime (p = 1.3e-4).

Authors
Wilson, JW; Degan, S; Gainey, CS; Deb, S; Dall, CP; Tameze-Rivas, Y; Zhang, J; Warren, WS
MLA Citation
Wilson, JW, Degan, S, Gainey, CS, Deb, S, Dall, CP, Tameze-Rivas, Y, Zhang, J, and Warren, WS. "In vivo pump-probe microscopy of melanoma: Characterizing shifts in excited state photodynamics with respect to invasiveness." January 1, 2015.
Source
scopus
Published In
Proceedings of SPIE
Volume
9329
Publish Date
2015
DOI
10.1117/12.2079886

Keratinocyte growth regulation TRP-ed up over downregulated TRPV4?

This commentary on an exciting new study (Fusi et al., 2014) puts the finding of TRPV4 downregulation in several nonmelanoma skin cancers into context. The original paper point toward possible use of TRPV4 as dermatopathologic marker, also toward the possibility that downregulated TRPV4 can affect biological properties of the cancer, by enhancing, but also regulating tumor growth. As calcium-permeable TRPV4 has recently been identified as UVB-receptor in skin keratinocytes, where it regulates skin tissue injury and pain after UVB overexposure, it is discussed whether TRPV4 downregulation can also be found in other non-UVB-exposed cancers.

Authors
Liedtke, W; Zhang, JY; Hall, RP; Steinhoff, M
MLA Citation
Liedtke, W, Zhang, JY, Hall, RP, and Steinhoff, M. "Keratinocyte growth regulation TRP-ed up over downregulated TRPV4?." The Journal of investigative dermatology 134.9 (September 2014): 2310-2312.
PMID
25120148
Source
epmc
Published In
Journal of Investigative Dermatology
Volume
134
Issue
9
Publish Date
2014
Start Page
2310
End Page
2312
DOI
10.1038/jid.2014.250

Effects of Y27632 on keratinocyte procurement and wound healing.

A number of Rho-kinase inhibitors have been developed for various clinical applications. We examined the effects of the Rho-kinase inhibitor Y27632 on keratinocyte proliferation and migration, and found that it promoted primary human keratinocyte proliferation and migration in both monolayer and skin explant cultures. In addition, topical application of Y27632 enhanced cutaneous wound closure in the majority of wounds in mice. The growth and migration effects of Y27632 appeared to be specific to keratinocytes compared with dermal fibroblasts, and required intact Jun kinase function. Y27632 seems to be a promising agent for keratinocyte procurement and wounding healing.

Authors
Gandham, VD; Maddala, RL; Rao, V; Jin, JY; Epstein, DL; Hall, RP; Zhang, JY
MLA Citation
Gandham, VD, Maddala, RL, Rao, V, Jin, JY, Epstein, DL, Hall, RP, and Zhang, JY. "Effects of Y27632 on keratinocyte procurement and wound healing." Clinical and experimental dermatology 38.7 (October 2013): 782-786.
PMID
23675999
Source
epmc
Published In
Clinical & Experimental Dermatology
Volume
38
Issue
7
Publish Date
2013
Start Page
782
End Page
786
DOI
10.1111/ced.12067

CYLD inhibits melanoma growth and progression through suppression of the JNK/AP-1 and β1-integrin signaling pathways.

The molecular mechanisms mediating cylindromatosis (CYLD) tumor suppressor function appear to be manifold. Here, we demonstrate that, in contrast to the increased levels of phosphorylated c-Jun NH(2)-terminal kinase (pJNK), CYLD was decreased in a majority of the melanoma cell lines and tissues examined. Exogenous expression of CYLD but not its catalytically deficient mutant markedly inhibited melanoma cell proliferation and migration in vitro and subcutaneous tumor growth in vivo. In addition, the melanoma cells expressing exogenous CYLD were unable to form pulmonary tumor nodules following tail-vein injection. At the molecular level, CYLD decreased β1-integrin and inhibited pJNK induction by tumor necrosis factor-α or cell attachment to collagen IV. Moreover, CYLD induced an array of other molecular changes associated with modulation of the "malignant" phenotype, including a decreased expression of cyclin D1, N-cadherin, and nuclear Bcl3, and an increased expression of p53 and E-cadherin. Most interestingly, coexpression of the constitutively active MKK7 or c-Jun mutants with CYLD prevented the above molecular changes, and fully restored melanoma growth and metastatic potential in vivo. Our findings demonstrate that the JNK/activator protein 1 signaling pathway underlies the melanoma growth and metastasis that are associated with CYLD loss of function. Thus, restoration of CYLD and inhibition of JNK and β1-integrin function represent potential therapeutic strategies for treatment of malignant melanoma.

Authors
Ke, H; Augustine, CK; Gandham, VD; Jin, JY; Tyler, DS; Akiyama, SK; Hall, RP; Zhang, JY
MLA Citation
Ke, H, Augustine, CK, Gandham, VD, Jin, JY, Tyler, DS, Akiyama, SK, Hall, RP, and Zhang, JY. "CYLD inhibits melanoma growth and progression through suppression of the JNK/AP-1 and β1-integrin signaling pathways." J Invest Dermatol 133.1 (January 2013): 221-229.
Website
http://hdl.handle.net/10161/15169
PMID
22832488
Source
pubmed
Published In
Journal of Investigative Dermatology
Volume
133
Issue
1
Publish Date
2013
Start Page
221
End Page
229
DOI
10.1038/jid.2012.253

CYLD inhibits melanoma growth and progression through suppression of the jnk/ap-1 and β1-integrin signaling pathways

The molecular mechanisms mediating cylindromatosis (CYLD) tumor suppressor function appear to be manifold. Here, we demonstrate that, in contrast to the increased levels of phosphorylated c-Jun NH 2 -terminal kinase (pJNK), CYLD was decreased in a majority of the melanoma cell lines and tissues examined. Exogenous expression of CYLD but not its catalytically deficient mutant markedly inhibited melanoma cell proliferation and migration in vitro and subcutaneous tumor growth in vivo. In addition, the melanoma cells expressing exogenous CYLD were unable to form pulmonary tumor nodules following tail-vein injection. At the molecular level, CYLD decreased β1-integrin and inhibited pJNK induction by tumor necrosis factor-α or cell attachment to collagen IV. Moreover, CYLD induced an array of other molecular changes associated with modulation of the "malignant" phenotype, including a decreased expression of cyclin D1, N-cadherin, and nuclear Bcl3, and an increased expression of p53 and E-cadherin. Most interestingly, coexpression of the constitutively active MKK7 or c-Jun mutants with CYLD prevented the above molecular changes, and fully restored melanoma growth and metastatic potential in vivo. Our findings demonstrate that the JNK/activator protein 1 signaling pathway underlies the melanoma growth and metastasis that are associated with CYLD loss of function. Thus, restoration of CYLD and inhibition of JNK and β1-integrin function represent potential therapeutic strategies for treatment of malignant melanoma. © 2013 The Society for Investigative Dermatology.

Authors
Ke, H; Augustine, CK; Gandham, VD; Jin, JY; Tyler, DS; Akiyama, SK; Hall, RP; Zhang, JY
MLA Citation
Ke, H, Augustine, CK, Gandham, VD, Jin, JY, Tyler, DS, Akiyama, SK, Hall, RP, and Zhang, JY. "CYLD inhibits melanoma growth and progression through suppression of the jnk/ap-1 and β1-integrin signaling pathways." Journal of Investigative Dermatology 133.1 (2013): 221-229.
Source
scival
Published In
Journal of Investigative Dermatology
Volume
133
Issue
1
Publish Date
2013
Start Page
221
End Page
229
DOI
10.1038/jid.2012.253

In vivo pump-probe microscopy of eumelanin, pheomelanin in melanoma

We employ pump-probe microscopy to highlight eumelanin versus pheomelanin content of pigmented skin lesions in vivo, and combine the technique with confocal reflectance and fluorescence microscopies to gain a more familiar illustration of the skin. © 2012.

Authors
Mitropoulos, T; Wilson, J; Degan, S; Selim, MA; Zhang, JY; Warrena, WS
MLA Citation
Mitropoulos, T, Wilson, J, Degan, S, Selim, MA, Zhang, JY, and Warrena, WS. "In vivo pump-probe microscopy of eumelanin, pheomelanin in melanoma." Biomedical Optics, BIOMED 2012 (December 1, 2012).
Source
scopus
Published In
Biomedical Optics, BIOMED 2012
Publish Date
2012

Abstract 381: Non-invasive histology for early detection of cutaneous melanoma and pigmented lesions in vivo

Authors
Degan, S; Wilson, JW; Mitropoulos, TE; Zhang, JY; Selim, MA; Warren, WS
MLA Citation
Degan, S, Wilson, JW, Mitropoulos, TE, Zhang, JY, Selim, MA, and Warren, WS. "Abstract 381: Non-invasive histology for early detection of cutaneous melanoma and pigmented lesions in vivo." Cancer Research 72.8 Supplement (April 15, 2012): 381-381.
Source
crossref
Published In
Cancer Research
Volume
72
Issue
8 Supplement
Publish Date
2012
Start Page
381
End Page
381
DOI
10.1158/1538-7445.AM2012-381

The role of the c-Jun N-terminal Kinase signaling pathway in skin cancer.

The c-Jun N-terminal Kinases (JNK), along with Erk and p38, constitute the principle members of the mitogen-activated protein kinase (MAPK) family. JNK functions primarily through AP1 family transcription factors to regulate a plethora of cellular processes, including cell proliferation, differentiation, survival and migration. It also cross-talks and integrates with other signaling pathways in a cell context-specific and cell type-specific manner. The current views of JNK function in various skin cancers and the need of developing JNK subunit-specific inhibitors for cancer type-specific applications have been summarized in this review.

Authors
Zhang, JY; Selim, MA
MLA Citation
Zhang, JY, and Selim, MA. "The role of the c-Jun N-terminal Kinase signaling pathway in skin cancer." American journal of cancer research 2.6 (January 2012): 691-698.
PMID
23226615
Source
epmc
Published In
American Journal of Cancer Research
Volume
2
Issue
6
Publish Date
2012
Start Page
691
End Page
698

In vivo pump-probe microscopy of melanoma and pigmented lesions

A growing number of dermatologists and pathologists are concerned that the rapidly rising incidence of melanoma reflects not a true 'epidemic' but an increasing tendency to overdiagnose pigmented lesions. Addressing this problem requires both a better understanding of early-stage melanoma and new diagnostic criteria based on more than just cellular morphology and architecture. Here we present a method for in-vivo optical microscopy that utilizes pump-probe spectroscopy to image the distribution of the two forms of melanin in skin: eumelanin and pheomelanin. Images are acquired in a scanning microscope with a sensitive modulation transfer technique by analyzing back-scattered probe light with a lock-in amplifier. Early-stage melanoma is studied in a human skin xenografted mouse model. Individual melanocytes have been observed, in addition to pigmented keratinocytes. Combining the pump-probe images simultaneously with other noninvasive laser microscopy methods (confocal reflectance, multiphoton autofluorescence, and second harmonic generation) allows visualization of the skin architecture, framing the functional pump-probe image in the context of the surrounding tissue morphology. It is found that pump-probe images of melanin can be acquired with low peak intensities, enabling wide field-of-view pigmentation surveys. Finally, we investigate the diagnostic potential of the additional chemical information available from pump-probe microscopy. © 2012 Copyright Society of Photo-Optical Instrumentation Engineers (SPIE).

Authors
Wilson, JW; Degan, S; Mitropoulos, T; Selim, MA; Zhang, JY; Warren, WS
MLA Citation
Wilson, JW, Degan, S, Mitropoulos, T, Selim, MA, Zhang, JY, and Warren, WS. "In vivo pump-probe microscopy of melanoma and pigmented lesions." Progress in Biomedical Optics and Imaging - Proceedings of SPIE 8226 (2012).
Source
scival
Published In
Proceedings of SPIE
Volume
8226
Publish Date
2012
DOI
10.1117/12.908821

Pump-probe melanoma imaging: Applications to high-resolution and in-vivo microscopy

Pump-probe imaging of melanin with near-infrared pulses has been extended to two new domains: high-resolution imaging of the melanin content of melanosomes in an individual melanocyte and epi-detected in vivo microscopy of a developing melanoma. © OSA/ CLEO 2011.

Authors
Wilson, JW; Matthews, TE; Degan, S; Zhang, JY; Simpson, MJ; Warren, WS
MLA Citation
Wilson, JW, Matthews, TE, Degan, S, Zhang, JY, Simpson, MJ, and Warren, WS. "Pump-probe melanoma imaging: Applications to high-resolution and in-vivo microscopy." Optics InfoBase Conference Papers (December 1, 2011).
Source
scopus
Published In
Optics InfoBase Conference Papers
Publish Date
2011

Pump-probe melanoma imaging: Applications to high-resolution and in-vivo microscopy

Pump-probe imaging of melanin with near-infrared pulses has been extended to two new domains: high-resolution imaging of the melanin content of melanosomes in an individual melanocyte and epi-detected in vivo microscopy of a developing melanoma. © OSA/ CLEO 2011.

Authors
Wilson, JW; Matthews, TE; Degan, S; Zhang, JY; Simpson, MJ; Warren, WS
MLA Citation
Wilson, JW, Matthews, TE, Degan, S, Zhang, JY, Simpson, MJ, and Warren, WS. "Pump-probe melanoma imaging: Applications to high-resolution and in-vivo microscopy." Optics InfoBase Conference Papers (December 1, 2011).
Source
scopus
Published In
Optics InfoBase Conference Papers
Publish Date
2011

Pump-probe microscopy captures cellular detail of melanoma in-vivo.

Pump-probe imaging of melanin with near-infrared pulses coupled with multphoton autofluorescence captures both chemical contrast and cellular detail in a live, developing melanoma. © 2011 OSA.

Authors
Wilson, JW; Matthews, TE; Degan, S; Zhang, JY; Simpson, MJ; Warren, WS
MLA Citation
Wilson, JW, Matthews, TE, Degan, S, Zhang, JY, Simpson, MJ, and Warren, WS. "Pump-probe microscopy captures cellular detail of melanoma in-vivo." Optics InfoBase Conference Papers (December 1, 2011).
Source
scopus
Published In
Optics InfoBase Conference Papers
Publish Date
2011

CYLD inhibits tumorigenesis and metastasis by blocking JNK/AP1 signaling at multiple levels.

CYLD has been recognized as a tumor suppressor due to its dominant genetic linkage to multiple types of epidermal tumors and a range of other cancers. The molecular mechanisms governing CYLD control of skin cancer are still unclear. Here, we showed that K14-driven epidermal expression of a patient-relevant and catalytically deficient CYLD truncated mutant (CYLD(m)) sensitized mice to skin tumor development in response to 7,12-dimethylbenz[α]anthracene (DMBA)/(12-O-tetradecanoylphorbol-13-acetate) TPA challenge. Tumors developed on transgenic mice were prone to malignant progression and lymph node metastasis and displayed increased activation of c-Jun-NH2-kinase (JNK) and the downstream c-Jun and c-Fos proteins. Most importantly, topical application of a pharmacologic JNK inhibitor significantly reduced tumor development and abolished metastasis in the transgenic mice. Further in line with these animal data, exogenous expression of CYLD(m) in A431, a human squamous cell carcinoma (SCC) cell line, markedly enhanced cell growth, migration, and subcutaneous tumor growth in an AP1-depdendent manner. In contrast, expression of the wild-type CYLD inhibited SCC tumorigenesis and AP1 function. Most importantly, CYLD(m) not only increased JNK activation but also induced an upregulation of K63 ubiquitination on both c-Jun and c-Fos, leading to sustained AP1 activation. Our findings uncovered c-Jun and c-Fos as novel CYLD targets and underscore that CYLD controls epidermal tumorigenesis through blocking the JNK/AP1 signaling pathway at multiple levels.

Authors
Miliani de Marval, P; Lutfeali, S; Jin, JY; Leshin, B; Selim, MA; Zhang, JY
MLA Citation
Miliani de Marval, P, Lutfeali, S, Jin, JY, Leshin, B, Selim, MA, and Zhang, JY. "CYLD inhibits tumorigenesis and metastasis by blocking JNK/AP1 signaling at multiple levels." Cancer Prev Res (Phila) 4.6 (June 2011): 851-859.
PMID
21478324
Source
pubmed
Published In
Cancer Prevention Research
Volume
4
Issue
6
Publish Date
2011
Start Page
851
End Page
859
DOI
10.1158/1940-6207.CAPR-10-0360

c-Jun promotes whereas JunB inhibits epidermal neoplasia.

Deregulation of the activator protein 1 (AP1) family gene regulators has been implicated in a wide range of diseases, including cancer. In this study we report that c-Jun was activated in human squamous cell carcinoma (SCC) and coexpression of c-Jun with oncogenic Ras was sufficient to transform primary human epidermal cells into malignancy in a regenerated human skin grafting model. In contrast, JunB was not induced in a majority of human SCC cells. Moreover, exogenous expression of JunB inhibited tumorigenesis driven by Ras or spontaneous human SCC cells. Conversely, the dominant-negative JunB mutant (DNJunB) promoted tumorigenesis, which is in contrast to the tumor-suppressor function of the corresponding c-Jun mutant. At the cellular level, JunB induced epidermal cell senescence and slowed cell growth in a cell-autonomous manner. Consistently, coexpression of JunB and Ras induced premature epidermal differentiation concomitant with upregulation of p16 and filaggrin and downregulation of cyclin D1 and cyclin-dependent kinase 4 (CDK4). These findings indicate that JunB and c-Jun differentially regulate cell growth and differentiation and induce opposite effects on epidermal neoplasia.JID JOURNAL CLUB ARTICLE: For questions, answers, and open discussion about this article, please go to http://www.nature.com/jid/journalclub.

Authors
Jin, JY; Ke, H; Hall, RP; Zhang, JY
MLA Citation
Jin, JY, Ke, H, Hall, RP, and Zhang, JY. "c-Jun promotes whereas JunB inhibits epidermal neoplasia." J Invest Dermatol 131.5 (May 2011): 1149-1158.
PMID
21289643
Source
pubmed
Published In
Journal of Investigative Dermatology
Volume
131
Issue
5
Publish Date
2011
Start Page
1149
End Page
1158
DOI
10.1038/jid.2011.1

In vivo and ex vivo epi-mode pump-probe imaging of melanin and microvasculature

We performed epi-mode pump-probe imaging of melanin in excised human pigmented lesions and both hemoglobin and melanin in live xenograft mouse melanoma models to depths greater than 100 μm. Eumelanin and pheomelanin images, which have been previously demonstrated to differentiate melanoma from benign lesions, were acquired at the dermal-epidermal junction with cellular resolution and modest optical powers (down to 15 mW). We imaged dermal microvasculature with the same wavelengths, allowing simultaneous acquisition of melanin, hemoglobin and multiphoton autofluorescence images. Molecular pump-probe imaging of melanocytes, skin structure and microvessels allows comprehensive, non-invasive characterization of pigmented lesions. © 2011 Optical Society of America.

Authors
Matthews, TE; Wilson, JW; Degan, S; Simpson, MJ; Jin, JY; Zhang, JY; Warren, WS
MLA Citation
Matthews, TE, Wilson, JW, Degan, S, Simpson, MJ, Jin, JY, Zhang, JY, and Warren, WS. "In vivo and ex vivo epi-mode pump-probe imaging of melanin and microvasculature." Biomedical Optics Express 2.6 (2011): 1576-1583.
Website
http://hdl.handle.net/10161/15168
PMID
21698020
Source
scival
Published In
Biomedical Optics Express
Volume
2
Issue
6
Publish Date
2011
Start Page
1576
End Page
1583
DOI
10.1364/BOE.2.001576

BCL2 interaction with actin in vitro may inhibit cell motility by enhancing actin polymerization

In addition to its well-defined role as an antagonist in apoptosis, we propose that BCL2 may act as an intracellular suppressor of cell motility and adhesion under certain conditions. Our evidence shows that, when overexpressed in both cancer and non-cancer cells, BCL2 can form a complex with actin and gelsolin that functions to decrease gelsolin-severing activity to increase actin polymerization and, thus, suppress cell adhesive processes. The linkage between increased BCL2 and increased actin polymerization on the one hand and suppression of cell adhesion, spreading and motility on the other hand, is a novel observation that may provide a plausible explanation for why BCL2 overexpression in some tumors is correlated with improved patient survival. In addition, we have identified conditions in vitro in which F-actin polymerization can be increased while cell motility is reduced. These findings underscore the possibility that BCL2 may be involved in modulating cytoskeleton reorganization and may provide an opportunity to explore signal transduction pathways important for cell adhesion and migration and to develop small molecule therapies for suppression of cancer metastasis. © 2011 Landes Bioscience.

Authors
Ke, H; Zhang, JY; Akiyama, SK; French, JE
MLA Citation
Ke, H, Zhang, JY, Akiyama, SK, and French, JE. "BCL2 interaction with actin in vitro may inhibit cell motility by enhancing actin polymerization." Cell Adhesion and Migration 5.1 (2011): 6-10.
PMID
20716950
Source
scival
Published In
Cell adhesion & migration
Volume
5
Issue
1
Publish Date
2011
Start Page
6
End Page
10
DOI
10.4161/cam.5.1.13175

The c-Jun NH2-terminal kinase 2 plays a dominant role in human epidermal neoplasia.

The c-Jun NH(2)-terminal kinase (JNK) signaling cascade has been implicated in a wide range of diseases, including cancer. It is unclear how different JNK proteins contribute to human cancer. Here, we report that JNK2 is activated in more than 70% of human squamous cell carcinoma (SCC) samples and that inhibition of JNK2 pharmacologically or genetically impairs tumorigenesis of human SCC cells. Most importantly, JNK2, but not JNK1, is sufficient to couple with oncogenic Ras to transform primary human epidermal cells into malignancy with features of SCC. JNK2 prevents Ras-induced cell senescence and growth arrest by reducing the expression levels of the cell cycle inhibitor p16 and the activation of NF-kappaB. On the other hand, JNK, along with phosphoinositide 3-kinase, is essential for Ras-induced glycolysis, an energy-producing process known to benefit cancer growth. These data indicate that JNK2 collaborates with other oncogenes, such as Ras, at multiple molecular levels to promote tumorigenesis and hence represents a promising therapeutic target for cancer.

Authors
Ke, H; Harris, R; Coloff, JL; Jin, JY; Leshin, B; Miliani de Marval, P; Tao, S; Rathmell, JC; Hall, RP; Zhang, JY
MLA Citation
Ke, H, Harris, R, Coloff, JL, Jin, JY, Leshin, B, Miliani de Marval, P, Tao, S, Rathmell, JC, Hall, RP, and Zhang, JY. "The c-Jun NH2-terminal kinase 2 plays a dominant role in human epidermal neoplasia." Cancer Res 70.8 (April 15, 2010): 3080-3088.
PMID
20354187
Source
pubmed
Published In
Cancer Research
Volume
70
Issue
8
Publish Date
2010
Start Page
3080
End Page
3088
DOI
10.1158/0008-5472.CAN-09-2923

BCL2 inhibits cell adhesion, spreading, and motility by enhancing actin polymerization

BCL2 is best known as a multifunctional anti-apoptotic protein. However, little is known about its role in cell-adhesive and motility events. Here, we show that BCL2 may play a role in the regulation of cell adhesion, spreading, and motility. When BCL2 was overexpressed in cultured murine and human cell lines, cell spreading, adhesion, and motility were impaired. Consistent with these results, the loss of Bcl2 resulted in higher motility observed in Bcl2-null mouse embryonic fibroblast (MEF) cells compared to wild type. The mechanism of BCL2 regulation of cell adhesion and motility may involve formation of a complex containing BCL2, actin, and gelsolin, which appears to functionally decrease the severing activity of gelsolin. We have observed that the lysate from MCF-7 and NIH3T3 cells that overexpressed BCL2 enhanced actin polymerization in cell-free in vitro assays. Confocal immunofluorescent localization of BCL2 and F-actin during spreading consistently showed that increased expression of BCL2 resulted in increased F-actin polymerization. Thus, the formation of BCL2 and gelsolin complexes (which possibly contain other proteins) appears to play a critical role in the regulation of cell adhesion and migration. Given the established correlation of cell motility with cancer metastasis, this result may explain why the expression of BCL2 in some tumor cell types reduces the potential for metastasis and is associated with improved patient prognosis. © 2010 IBCB, SIBS, CAS All rights reserved.

Authors
Ke, H; Parron, VI; Reece, J; Zhang, JY; Akiyama, SK; French, JE
MLA Citation
Ke, H, Parron, VI, Reece, J, Zhang, JY, Akiyama, SK, and French, JE. "BCL2 inhibits cell adhesion, spreading, and motility by enhancing actin polymerization." Cell Research 20.4 (2010): 458-469.
Website
http://hdl.handle.net/10161/15167
PMID
20142842
Source
scival
Published In
Cell Research
Volume
20
Issue
4
Publish Date
2010
Start Page
458
End Page
469
DOI
10.1038/cr.2010.21

Tumor necrosis factor receptor 1/c-Jun-NH2-kinase signaling promotes human neoplasia.

The tumor necrosis factor alpha receptor (TNFR1) activates downstream effectors that include the mitogen-activated protein kinase kinase 7 (MKK7)/c-Jun-NH(2)-kinase (JNK)/activator protein 1 (AP1) cascade. Here, we report that JNK is activated in a majority of spontaneous human squamous cell carcinomas (SCC). JNK pathway induction bypassed cell cycle restraints induced by oncogenic Ras and cooperated with Ras to convert normal human epidermis into tumors indistinguishable from SCC, confirming its oncogenic potency in human tissue. Inhibiting MKK7, JNK, and AP1 as well as TNFR1 itself using genetic, pharmacologic, or antibody-mediated approaches abolished invasive human epidermal neoplasia in a tumor cell autonomous fashion. The TNFR1/MKK7/JNK/AP1 cascade thus promotes human neoplasia and represents a potential therapeutic target for human epithelial cancers.

Authors
Zhang, JY; Adams, AE; Ridky, TW; Tao, S; Khavari, PA
MLA Citation
Zhang, JY, Adams, AE, Ridky, TW, Tao, S, and Khavari, PA. "Tumor necrosis factor receptor 1/c-Jun-NH2-kinase signaling promotes human neoplasia." Cancer Res 67.8 (April 15, 2007): 3827-3834.
PMID
17440097
Source
pubmed
Published In
Cancer Research
Volume
67
Issue
8
Publish Date
2007
Start Page
3827
End Page
3834
DOI
10.1158/0008-5472.CAN-06-4017

Motif module map reveals enforcement of aging by continual NF-κB activity

Aging is characterized by specific alterations in gene expression, but their underlying mechanisms and functional consequences are not well understood. Here we develop a systematic approach to identify combinatorial cis-regulatory motifs that drive age-dependent gene expression across different tissues and organisms. Integrated analysis of 365 microarrays spanning nine tissue types predicted fourteen motifs as major regulators of age-dependent gene expression in human and mouse. The motif most strongly associated with aging was that of the transcription factor NF-κB. Inducible genetic blockade of NF-κB for 2 wk in the epidermis of chronologically aged mice reverted the tissue characteristics and global gene expression programs to those of young mice. Age-specific NF-κB blockade and orthogonal cell cycle interventions revealed that NF-κB controls cell cycle exit and gene expression signature of aging in parallel but not sequential pathways. These results identify a conserved network of regulatory pathways underlying mammalian aging and show that NF-κB is continually required to enforce many features of aging in a tissue-specific manner. © 2007 by Cold Spring Harbor Laboratory Press.

Authors
Adler, AS; Sinha, S; Kawahara, TLA; Zhang, JY; Segal, E; Chang, HY
MLA Citation
Adler, AS, Sinha, S, Kawahara, TLA, Zhang, JY, Segal, E, and Chang, HY. "Motif module map reveals enforcement of aging by continual NF-κB activity." Genes and Development 21.24 (2007): 3244-3257.
PMID
18055696
Source
scival
Published In
Genes & development
Volume
21
Issue
24
Publish Date
2007
Start Page
3244
End Page
3257
DOI
10.1101/gad.1588507

CDK4 regulation by TNFR1 and JNK is required for NF-kappaB-mediated epidermal growth control.

Nuclear factor kappaB (NF-kappaB) mediates homeostatic growth inhibition in the epidermis, and a loss of NF-kappaB function promotes proliferation and oncogenesis. To identify mechanisms responsible for these effects, we impaired NF-kappaB action in the epidermis by three different genetic approaches, including conditional NF-kappaB blockade. In each case, epidermal hyperplasia was accompanied by an increase in both protein levels and tissue distribution of the G1 cell cycle kinase, CDK4. CDK4 up-regulation required intact TNFR1 and c-Jun NH2-terminal kinase (JNK) function. Cdk4 gene deletion concomitant with conditional NF-kappaB blockade demonstrated that CDK4 is required for growth deregulation. Therefore, epidermal homeostasis depends on antagonist regulation of CDK4 expression by NF-kappaB and TNFR1/JNK.

Authors
Zhang, JY; Tao, S; Kimmel, R; Khavari, PA
MLA Citation
Zhang, JY, Tao, S, Kimmel, R, and Khavari, PA. "CDK4 regulation by TNFR1 and JNK is required for NF-kappaB-mediated epidermal growth control." J Cell Biol 168.4 (February 14, 2005): 561-566.
PMID
15699216
Source
pubmed
Published In
The Journal of Cell Biology
Volume
168
Issue
4
Publish Date
2005
Start Page
561
End Page
566
DOI
10.1083/jcb.200411060

NF-kappaB RelA opposes epidermal proliferation driven by TNFR1 and JNK.

NF-kappaB inhibition promotes epidermal tumorigenesis; however, whether this reflects an underlying role in homeostasis or a special case confined to neoplasia is unknown. Embryonic lethality of mice lacking NF-kappaB RelA has hindered efforts to address this. We therefore generated developmentally mature RelA(-/-) skin. RelA(-/-) epidermis displays hyperplasia without abnormal differentiation, inflammation, or apoptosis. Hyperproliferation is TNFR1-dependent because Tnfr1 deletion normalized cell division. TNFR1-dependent JNK activation occurred in RelA(-/-) epidermis, and JNK inhibition abolished hyperproliferation due to RelA deficiency. Thus, RelA antagonizes TNFR1-JNK proliferative signals in epidermis and plays a nonredundant role in restraining epidermal growth.

Authors
Zhang, JY; Green, CL; Tao, S; Khavari, PA
MLA Citation
Zhang, JY, Green, CL, Tao, S, and Khavari, PA. "NF-kappaB RelA opposes epidermal proliferation driven by TNFR1 and JNK." Genes Dev 18.1 (January 1, 2004): 17-22.
PMID
14724177
Source
pubmed
Published In
Genes & development
Volume
18
Issue
1
Publish Date
2004
Start Page
17
End Page
22
DOI
10.1101/gad.1160904

Erratum: NF-κB RelA opposes epidermal proliferation driven by TNFR1 and JNK (Genes and Development (2004) 18 (17-22))

Authors
Zhang, JY; Green, CL; Tao, S; Khavari, PA
MLA Citation
Zhang, JY, Green, CL, Tao, S, and Khavari, PA. "Erratum: NF-κB RelA opposes epidermal proliferation driven by TNFR1 and JNK (Genes and Development (2004) 18 (17-22))." Genes and Development 18.4 (2004): 461--.
Source
scival
Published In
Genes and Development
Volume
18
Issue
4
Publish Date
2004
Start Page
461-

Innate inhibition of adaptive immunity: Mycobacterium tuberculosis-induced IL-6 inhibits macrophage responses to IFN-gamma.

In humans and in mice, control of the intracellular pathogen, Mycobacterium tuberculosis (Mtb), requires IFN-gamma. Although the adaptive immune response results in production of substantial amounts of IFN-gamma in response to Mtb, the immune response is unable to eradicate the infection in most cases. We have previously reported evidence that Mtb inhibits macrophage responses to IFN-gamma, suggesting that this may limit the ability of IFN-gamma to stimulate macrophages to kill Mtb. We have also observed that uninfected macrophages, adjacent to infected macrophages in culture, exhibit decreased responses to IFN-gamma. Here we report that IL-6 secreted by Mtb-infected macrophages inhibits the responses of uninfected macrophages to IFN-gamma. IL-6 selectively inhibits a subset of IFN-gamma-responsive genes at the level of transcriptional activation without inhibiting activation or function of STAT1. Inhibition of macrophage responses to IFN-gamma by IL-6 requires new protein synthesis, but this effect is not attributable to suppressor of cytokine signaling 1 or 3. These results reveal a novel function for IL-6 and indicate that IL-6 secreted by Mtb-infected macrophages may contribute to the inability of the cellular immune response to eradicate infection.

Authors
Nagabhushanam, V; Solache, A; Ting, L-M; Escaron, CJ; Zhang, JY; Ernst, JD
MLA Citation
Nagabhushanam, V, Solache, A, Ting, L-M, Escaron, CJ, Zhang, JY, and Ernst, JD. "Innate inhibition of adaptive immunity: Mycobacterium tuberculosis-induced IL-6 inhibits macrophage responses to IFN-gamma." J Immunol 171.9 (November 1, 2003): 4750-4757.
PMID
14568951
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
171
Issue
9
Publish Date
2003
Start Page
4750
End Page
4757

Divergent gene regulation and growth effects by NF-kappa B in epithelial and mesenchymal cells of human skin.

NF-kappa B regulates normal and pathological processes, including neoplasia, in a tissue-context-dependent manner. In skin, NF-kappa B is implicated in epidermal homeostasis as well as in the pathogenesis of squamous cell carcinoma; however, its function in the underlying mesenchymal dermis has been unclear. To gain insight into NF-kappa B roles in these two adjacent cutaneous tissue compartments, NF-kappa B effects on expression of 12 435 genes were determined in epidermal keratinocytes and dermal fibroblasts. Although NF-kappa B induced proinflammatory and antiapoptotic genes in both settings, it exhibited divergent effects on growth regulatory genes. In keratinocytes, but not in fibroblasts, NF-kappa B induced p21(CIP1), which was sufficient to inhibit growth of both cell types. Levels of growth inhibitory factor (GIF), in contrast, were increased by NF-kappa B in both settings but inhibited growth only in keratinocytes. These findings indicate that transcription factors such as NF-kappa B can program tissue-selective effects via both differential target gene induction as well as by inducing common targets that exert differing effects depending on cellular lineage.

Authors
Hinata, K; Gervin, AM; Jennifer Zhang, Y; Khavari, PA
MLA Citation
Hinata, K, Gervin, AM, Jennifer Zhang, Y, and Khavari, PA. "Divergent gene regulation and growth effects by NF-kappa B in epithelial and mesenchymal cells of human skin." Oncogene 22.13 (April 3, 2003): 1955-1964.
PMID
12673201
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
22
Issue
13
Publish Date
2003
Start Page
1955
End Page
1964
DOI
10.1038/sj.onc.1206198

Escaping G1 restraints on neoplasia--Cdk4 regulation by Ras and NF-kappa B.

Authors
Lazarov, M; Green, CL; Zhang, JY; Kubo, Y; Dajee, M; Khavari, PA
MLA Citation
Lazarov, M, Green, CL, Zhang, JY, Kubo, Y, Dajee, M, and Khavari, PA. "Escaping G1 restraints on neoplasia--Cdk4 regulation by Ras and NF-kappa B." Cell Cycle 2.2 (March 2003): 79-80.
PMID
12695650
Source
pubmed
Published In
Cell Cycle
Volume
2
Issue
2
Publish Date
2003
Start Page
79
End Page
80

NF-kappaB blockade and oncogenic Ras trigger invasive human epidermal neoplasia.

The nuclear factor NF-kappaB and oncogenic Ras can alter proliferation in epidermis, the most common site of human cancer. These proteins are implicated in epidermal squamous cell carcinoma in mice, however, the potential effects of altering their function are uncertain. Whereas inhibition of NF-kappaB enhances apoptosis in certain tumours, blockade of NF-kappaB predisposes murine skin to squamous cell carcinoma. Because therapeutics inhibiting Ras and NF-kappaB pathways are being developed to treat human cancer, it is essential to assess the effects of altering these regulators. The medical relevance of murine studies is limited, however, by differences between mouse and human skin, and by the greater ease of transforming murine cells. Here we show that in normal human epidermal cells both NF-kappaB and oncogenic Ras trigger cell-cycle arrest. Growth arrest triggered by oncogenic Ras can be bypassed by IkappaBalpha-mediated blockade of NF-kappaB, generating malignant human epidermal tissue resembling squamous cell carcinoma. Human cell tumorigenesis is dependent on laminin 5 and alpha6beta4 integrin. Thus, IkappaBalpha circumvents restraints on growth promotion induced by oncogenic Ras and can act with Ras to induce invasive human tissue neoplasia.

Authors
Dajee, M; Lazarov, M; Zhang, JY; Cai, T; Green, CL; Russell, AJ; Marinkovich, MP; Tao, S; Lin, Q; Kubo, Y; Khavari, PA
MLA Citation
Dajee, M, Lazarov, M, Zhang, JY, Cai, T, Green, CL, Russell, AJ, Marinkovich, MP, Tao, S, Lin, Q, Kubo, Y, and Khavari, PA. "NF-kappaB blockade and oncogenic Ras trigger invasive human epidermal neoplasia." Nature 421.6923 (February 6, 2003): 639-643.
PMID
12571598
Source
pubmed
Published In
Nature
Volume
421
Issue
6923
Publish Date
2003
Start Page
639
End Page
643
DOI
10.1038/nature01283

A linking function for the cellulose-binding protein SP85 in the spore coat of Dictyostelium discoideum.

SP85 is a multidomain protein of the Dictyostelium spore coat whose C-terminal region binds cellulose in vitro. To map domains critical for localizing SP85 and for binding to other proteins in vivo, its N- and C-terminal regions, and a hybrid fusion of the N- and C-regions, were expressed in prespore cells. Immunofluorescence showed that only the N-terminal region and the N/C-hybrid accumulated in prespore vesicles, where coat proteins are normally stored prior to secretion. In contrast, only the C-terminal region and N/C-hybrid were incorporated into the coat after secretion. To determine if SP85 is important for the incorporation of other coat proteins, an SP85-null strain was created and found to mislocalize the coat protein SP65 to the interspore matrix. In vitro binding studies demonstrated that the SP85 C-terminal region bound SP65 and cellulose simultaneously, and SP65 incorporation was rescued in vivo by the C-terminal region. SP85-null spores showed increased latent permeability to a fluorescent lectin probe and accelerated germination times, and decreased buoyant density of their coats, suggesting that coat barrier functions were compromised. Dominant negative reductions in barrier functions also resulted from expression of the SP85 terminal regions, suggesting that a linking activity was important for SP85's function. Thus, separate domains of SP85 specify prespore vesicle compartmentalization and coat incorporation, and additional domains link SP65 to the coat and simultaneously interact with other binding partners which contribute to coat barrier functions.

Authors
Zhang, Y; Zhang, P; West, CM
MLA Citation
Zhang, Y, Zhang, P, and West, CM. "A linking function for the cellulose-binding protein SP85 in the spore coat of Dictyostelium discoideum." J Cell Sci 112 ( Pt 23) (December 1999): 4367-4377.
PMID
10564654
Source
pubmed
Published In
Journal of cell science
Volume
112 ( Pt 23)
Publish Date
1999
Start Page
4367
End Page
4377

Two proteins of the Dictyostelium spore coat bind to cellulose in vitro.

The spore coat of Dictyostelium contains nine different proteins and cellulose. Interactions between protein and cellulose were investigated using an in vitro binding assay. Proteins extracted from coats with urea and 2-mercaptoethanol could, after removal of urea by gel filtration, efficiently bind to particles of cellulose (Avicel), but not Sephadex or Sepharose. Two proteins, SP85 and SP35, were enriched in the reconstitution, and they retained their cellulose binding activities after purification by ion exchange chromatography under denaturing conditions to suppress protein--protein interactions. Neither protein exhibited cellulase activity, though under certain conditions SP85 copurified with a cellulase activity which appeared after germination. Amino acid sequencing indicated that SP85 and SP35 are encoded by the previously described pspB and psvA genes. This was confirmed for SP85 by showing that natural M(r) polymorphisms correlated with changes in the number of tetrapeptide-encoding sequence repeats in pspB. Using PCR to reconstruct missing elements from the recombinogenic middle region of pspB, SP85 was shown to consist of three sequence domains separated by two groups of the tetrapeptide repeats. Expression of partial pspB cDNAs in Escherichia coli showed that cellulose-binding activity resided in the Cys-rich COOH-terminal domain of SP85. This cellulose-binding activity can explain SP85's ultrastructural colocalization with cellulose in vivo. Amino acid composition and antibody binding data showed that SP35 is derived from the Cys-rich N-terminal region of the previously described psvA protein. SP85 and SP35 may link other proteins to cellulose during coat assembly and germination.

Authors
Zhang, Y; Brown, RD; West, CM
MLA Citation
Zhang, Y, Brown, RD, and West, CM. "Two proteins of the Dictyostelium spore coat bind to cellulose in vitro." Biochemistry 37.30 (July 28, 1998): 10766-10779.
PMID
9692967
Source
pubmed
Published In
Biochemistry
Volume
37
Issue
30
Publish Date
1998
Start Page
10766
End Page
10779
DOI
10.1021/bi9808013

SP75 is encoded by the DP87 gene and belongs to a family of modular Dictyostelium discoideum outer layer spore coat proteins.

Highly purified spore coats of Dictyostelium discoideum each contained about 5 x 10(6) protein molecules as determined by amino acid composition analysis. By two-dimensional gel electrophoresis the coats were found to contain nine major-abundance and numerous minor protein species, most of which were highly enriched relative to the adjacent interspore matrix. Protein was nearly quantitatively eluted by denaturants and 2-mercaptoethanol, showing that it was not irreversibly cross-linked. Because a reducing agent is required together with denaturants to elute most proteins if their free thiol groups have been prealkylated, it was concluded that the D. discoideum spore coat proteins are disulfide cross-linked into the matrix. One major coat protein, SP75, was partially sequenced and found to be encoded by the previously identified DP87 gene; this finding was supported by additional physical, genetic, biochemical and microscopic evidence. The five major proteins for which genes have been cloned were associated with the outer layer of the coat. In coats missing one or more of four of these proteins as a result of gene disruption, there were physical changes but, with one exception, the other major coat proteins appeared to be incorporated normally. Sequence analysis showed that these five outer layer coat proteins are homologous and consist of alternating sequence motifs related to epithelial mucin repeats, basic proline repeats found in salivary acidic proline-rich proteins, the NH2-terminal subdomain of epidermal growth factor modules and other cysteine repeats. Based on these and other observations, outer layer coat proteins are predicted to organize indeterminately to form a cell surface microenvironment supportive of cellulose morphogenesis during spore coat formation.

Authors
West, CM; Mao, J; van der Wel, H; Erdos, GW; Zhang, Y
MLA Citation
West, CM, Mao, J, van der Wel, H, Erdos, GW, and Zhang, Y. "SP75 is encoded by the DP87 gene and belongs to a family of modular Dictyostelium discoideum outer layer spore coat proteins." Microbiology 142 ( Pt 8) (August 1996): 2227-2243.
PMID
8760935
Source
pubmed
Published In
Microbiology (Reading, England)
Volume
142 ( Pt 8)
Publish Date
1996
Start Page
2227
End Page
2243
DOI
10.1099/13500872-142-8-2227
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