Abstract

? The major objective of this grant over the past 20 years has been to study key aspects of the mechanisms of immunologic cytotoxicity in skin disease. The major focus has been on cytotoxicity to melanocytes and keratinocytes in diseases such as vitiligo and lupus erythematosus. The purpose of this COMPETITIVE RENEWAL is to characterize the mechanisms of Fas-mediated cytotoxicity in melanocytes and basal keratinocytes, especially in situations where the normal resistance of these cells is overcome. This grant will emphasize studies to identify the anti-apoptotic defenses of these cells, and to characterize the signaling pathways that maintain these defenses. This proposal addresses the molecular and cellular biology of a fundamental characteristic of the cells of the basal layer of the epidermis: their intrinsic resistance to immunologic cytotoxicity, especially Fas-mediated apoptosis:
Specific Aim 1 : To confirm that both melanocytes and proliferating keratinocytes are Fas Type II cells, requiring both extrinsic and intrinsic pathways for Fas-mediated apoptosis.
Specific Aim 2, 3 and 4: To demonstrate the importance of PI3K/Akt, NF-kB, and Ras pathways in control of the susceptibility of melanocytes and keratinocytes to Fas-mediated apoptosis, and identify the anti-apoptotic targets involved in extrinsic and/or intrinsic pathways for Fas-mediated apoptosis.
Specific Aim 2 : For the PI3K/Akt pathway.
Specific Aim 3 : For the NF-kB pathway.
Specific Aim 4 : For the Ras pathway.
Specific Aim 5 : To identify unanticipated anti-apoptotic targets of PI3K, NF-kB, and Ras for Fas mediated apoptosis in melanocytes and basal keratinocytes using proteomic analysis. In summary, we will demonstrate that melanocytes and keratinocytes require the intrinsic pathway for Fas-mediated apoptosis (aim 1), and we will determine whether and how PI3K/Akt, NF-kB, and Ras control the resistance of melanocytes and keratinocytes to Fas-mediated apoptosis (aim 2, 3, and 4). In addition, we will identify unanticipated targets for these signal pathways that contribute to anti-apoptotic defenses in these cells (aim 5). ? ?

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR026427-22
Application #
6793963
Study Section
Special Emphasis Panel (ZRG1-GMA-1 (01))
Program Officer
Moshell, Alan N
Project Start
1980-01-01
Project End
2008-08-31
Budget Start
2004-09-01
Budget End
2005-08-31
Support Year
22
Fiscal Year
2004
Total Cost
$358,882
Indirect Cost

  Institution

Name
University of Colorado Denver
Department
Dermatology
Type
Schools of Medicine
DUNS #
041096314
City
Aurora
State
CO
Country
United States
Zip Code
80045

  Publications

Reuland, Steven N; Smith, Shilo M; Bemis, Lynne T et al. (2013) MicroRNA-26a is strongly downregulated in melanoma and induces cell death through repression of silencer of death domains (SODD). J Invest Dermatol 133:1286-93
Reuland, Steven N; Goldstein, Nathaniel B; Partyka, Katie A et al. (2012) ABT-737 synergizes with Bortezomib to kill melanoma cells. Biol Open 1:92-100
Reuland, Steven N; Goldstein, Nathaniel B; Partyka, Katie A et al. (2011) The combination of BH3-mimetic ABT-737 with the alkylating agent temozolomide induces strong synergistic killing of melanoma cells independent of p53. PLoS One 6:e24294
Smith, Shilo M; Wunder, Michael B; Norris, David A et al. (2011) A simple protocol for using a LDH-based cytotoxicity assay to assess the effects of death and growth inhibition at the same time. PLoS One 6:e26908
Goldstein, Nathaniel B; Johannes, Widya U; Gadeliya, Agnessa V et al. (2009) Active N-Ras and B-Raf inhibit anoikis by downregulating Bim expression in melanocytic cells. J Invest Dermatol 129:432-7
Miller, Leslie A; Goldstein, Nathaniel B; Johannes, Widya U et al. (2009) BH3 mimetic ABT-737 and a proteasome inhibitor synergistically kill melanomas through Noxa-dependent apoptosis. J Invest Dermatol 129:964-71
Shellman, Yiqun G; Howe, William R; Miller, Leslie A et al. (2008) Hyperthermia induces endoplasmic reticulum-mediated apoptosis in melanoma and non-melanoma skin cancer cells. J Invest Dermatol 128:949-56
Ruth, Mariah C; Xu, Yisheng; Maxwell, Ian H et al. (2006) RhoC promotes human melanoma invasion in a PI3K/Akt-dependent pathway. J Invest Dermatol 126:862-8
Shellman, Yiqun G; Makela, Marja; Norris, David A (2006) Induction of secreted matrix metalloproteinase-9 activity in human melanoma cells by extracellular matrix proteins and cytokines. Melanoma Res 16:207-11
Ribble, Deborah; Goldstein, Nathaniel B; Norris, David A et al. (2005) A simple technique for quantifying apoptosis in 96-well plates. BMC Biotechnol 5:12

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