Abstract

The objectives of this proposal are to determine how protein kinase Cepsilon (PKCepsilon) signals tumor necrosis factor alpha (TNFalpha) release and whether this TNFalpha release mediates the development of metastatic squamous cell carcinoma (mSCC) in PKCepsilon-overexpressing transgenic mice. PKC is not only the major intracellular receptor for the mouse skin tumor promoter 12-0-tetradecanoylphorbol-13-acetate (TPA) but also is activated by a variety of stress factors including ultraviolet irradiation (UVR). PKCepsilon is among the six PKC isoforms (alpha, delta, epsilon, eta, mu, delta) expressed in the mouse skin. To determine the in vivo functional specificity of PKCe in mouse skin carcinogenesis, we generated PKCepsilon transgenic mouse (FVB/N) lines 224 and 215 that overexpress approximately 8- and 18-fold respectively PKCepsilon protein over endogenous levels in basal epidermal cells. PKCepsilon transgenic mice were observed to be highly sensitive to the development of mSCC elicited either by the repeated exposure to UVR or by the DMBA (100 nmol) - TPA (5nmol) tumor promotion protocol. During studies to find clues about how PKCe overexpression mediates development of mSCC, we found that PKCepsilon transgenic mice have dramatically elevated TNFalpha serum levels relative to their wild-type littermates either following UVR exposure or during skin tumor promotion by TPA. TPA-stimulated serum TNFalpha levels were proportional to the level of expression of epidermal PKCe in transgenic mouse lines 224 and 215. Preliminary results indicate that mouse keratinocytes are the primary source of serum TNFalpha levels, and TPA affects ectodomain shedding of TNFalpha. The release of soluble TNFalpha is catalyzed by the TNFalpha converting enzyme (TACE), a transmembrane metalloproteinase. We hypothesize that: 1) UVR- or TPA-stimulated TNFalpha release is the key downstream component of the PKCepsilon signal transduction pathway to development of mSCC in PKCe transgenic mice and 2) PKCepsilon mediates UVR/TPA-stimulated TNFalpha release by regulating TACE activity. We propose two specific aims to test this hypothesis:
Specific Aim #1 : To determine whether TNFalpha is essential for the development of mSCC in PKCepsilon transgenic mice.
Specific Aim #2 : To determine how PKCe mediates UVR- or TPA-stimulated release of TNFalpha.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA102431-05
Application #
7339324
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Poland, Alan P
Project Start
2004-04-12
Project End
2009-02-28
Budget Start
2008-03-01
Budget End
2009-02-28
Support Year
5
Fiscal Year
2008
Total Cost
$254,537
Indirect Cost

  Institution

Name
University of Wisconsin Madison
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715

  Publications

Hafeez, Bilal Bin; Meske, Louise; Singh, Ashok et al. (2016) Tissue-specific conditional PKC? knockout mice: a model to precisely reveal PKC? functional role in initiation, promotion and progression of cancer. Oncotarget 7:33069-80
Singh, Ashok; Willems, Estelle; Singh, Anupama et al. (2016) Ultraviolet radiation-induced tumor necrosis factor alpha, which is linked to the development of cutaneous SCC, modulates differential epidermal microRNAs expression. Oncotarget 7:17945-56
Singh, Ashok; Willems, Estelle; Singh, Anupama et al. (2016) Ultraviolet radiation-induced differential microRNA expression in the skin of hairless SKH1 mice, a widely used mouse model for dermatology research. Oncotarget 7:84924-84937
Singh, Ashok; Singh, Anupama; Bauer, Samuel J et al. (2016) Genetic deletion of TNF? inhibits ultraviolet radiation-induced development of cutaneous squamous cell carcinomas in PKC? transgenic mice via inhibition of cell survival signals. Carcinogenesis 37:72-80
Singh, Anupama; Singh, Ashok; Sand, Jordan M et al. (2015) Topically applied Hsp90 inhibitor 17AAG inhibits UVR-induced cutaneous squamous cell carcinomas. J Invest Dermatol 135:1098-1107
Singh, Ashok; Singh, Anupama; Sand, Jordan M et al. (2013) Protein Kinase C ? , Which Is Linked to Ultraviolet Radiation-Induced Development of Squamous Cell Carcinomas, Stimulates Rapid Turnover of Adult Hair Follicle Stem Cells. J Skin Cancer 2013:452425
Sand, Jordan Marshall; Bin Hafeez, Bilal; Aziz, Moammir Hasan et al. (2012) Ultraviolet radiation and 12-O-tetradecanoylphorbol-13-acetate-induced interaction of mouse epidermal protein kinase Cýý with Stat3 involve integration with ERK1/2. Mol Carcinog 51:291-302
Aziz, Moammir Hasan; Sundling, Kaitlin Elizabeth; Dreckschmidt, Nancy Ellen et al. (2009) Protein kinase Cepsilon inhibits UVR-induced expression of FADD, an adaptor protein, linked to both Fas- and TNFR1-mediated apoptosis. J Invest Dermatol 129:2011-21
Aziz, Moammir H; Manoharan, Herbert T; Sand, Jordan M et al. (2007) Protein kinase Cepsilon interacts with Stat3 and regulates its activation that is essential for the development of skin cancer. Mol Carcinog 46:646-53
Aziz, Moammir H; Manoharan, Herbert T; Verma, Ajit K (2007) Protein kinase C epsilon, which sensitizes skin to sun's UV radiation-induced cutaneous damage and development of squamous cell carcinomas, associates with Stat3. Cancer Res 67:1385-94

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