Project 3 utilizes human and murine model systems to address the links between signature genetic events in melanoma progression (p16INK4a loss and N-RAS/B-RAF mutation) and the DMAdamage response in melanocytes. This project combines analyses of novel murine models of melanoma with a comprehensive molecular and immunohistochemical study of a large, clinically annotated human melanoma database.
In specific aim 1, we combine two different RAS alleles, conditionally inactivatable p53 and p16INK4a alleles, and an inducible, melanocyte-specific CRE allele to produce new murine models of melanoma that are highly faithful to the human genetics of this tumor. In these models, all oncogenic events are restricted to the melanocytic compartment, and include somatic inactivation of p16/p53 or somatic activation of K-RAS, Two of the alleles (Tyr-CRE-ER-T and conditional p16INK4a) are newly characterized and unpublished.
In specific aim 2, we combine these novel murine models of melanoma with neonatal UV-B exposure to facilitate in vivo melanomagenesis.
Specific aim 2 also includes a detailed immunohistochemical analysis of the kinetics of the expression of markers of the DNA damage response and senescence with or without UV- B treatment in the setting of RAS activation, p16INK4a loss and/or p53 loss.
This specific aim employs a similar approach to analyze human dermal reconstructs in immunodeficient mice with and without UV exposure using reagents supplied from projects 1 and 2.
In specific aim 3, we extend our analysis of human primary formalin-fixed and paraffin-embedded melanocytic lesions to identify the relationship among RAS/RAF/p16INK4a mutation, ERK MAP kinase activation, senescence and the DNA damage response in the progression from nevus to metastatic tumor.
This specific aim i ncludes a comprehensive immunohistochemical analysis of several markers of the DNA damage response and senescence, as well as a mutational analysis of N-RAS, B-RAF and p16INK4a.
This specific aim i s powered (250 melanocytic lesions from nevus to metaststic melanoma) to account for molecular heterogeneity in primary tumors but still uncover biologically significant relationships among these signature genetic events, the DNA damage response and melanoma progression. We expect this work will further our basic understanding of human melanoma progression as well as identify new clinjcal predictors of disease progression and outcome.
| Cordeiro-Stone, Marila; McNulty, John J; Sproul, Christopher D et al. (2016) Effective intra-S checkpoint responses to UVC in primary human melanocytes and melanoma cell lines. Pigment Cell Melanoma Res 29:68-80 |
| Kaufmann, William K; Carson, Craig C; Omolo, Bernard et al. (2014) Mechanisms of chromosomal instability in melanoma. Environ Mol Mutagen 55:457-71 |
| Chen, Liddy M; Ibrahim, Joseph G; Chu, Haitao (2014) Flexible stopping boundaries when changing primary endpoints after unblinded interim analyses. J Biopharm Stat 24:817-33 |
| Lin, Ja-An; Zhu, Hongtu; Mihye, Ahn et al. (2014) Functional-mixed effects models for candidate genetic mapping in imaging genetic studies. Genet Epidemiol 38:680-91 |
| Nikolaishvilli-Feinberg, Nana; Cohen, Stephanie M; Midkiff, Bentley et al. (2014) Development of DNA damage response signaling biomarkers using automated, quantitative image analysis. J Histochem Cytochem 62:185-96 |
| Kricker, Anne; Armstrong, Bruce K; Goumas, Chris et al. (2013) Survival for patients with single and multiple primary melanomas: the genes, environment, and melanoma study. JAMA Dermatol 149:921-7 |
| Schlegel, Jennifer; Sambade, Maria J; Sather, Susan et al. (2013) MERTK receptor tyrosine kinase is a therapeutic target in melanoma. J Clin Invest 123:2257-67 |
| Omolo, Bernard; Carson, Craig; Chu, Haitao et al. (2013) A prognostic signature of G(2) checkpoint function in melanoma cell lines. Cell Cycle 12:1071-82 |
| Smith-Roe, Stephanie L; Patel, Shivani S; Zhou, Yingchun et al. (2013) Separation of intra-S checkpoint protein contributions to DNA replication fork protection and genomic stability in normal human fibroblasts. Cell Cycle 12:332-45 |
| Lakhter, Alexander J; Sahu, Ravi P; Sun, Yang et al. (2013) Chloroquine promotes apoptosis in melanoma cells by inhibiting BH3 domain-mediated PUMA degradation. J Invest Dermatol 133:2247-54 |
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