BLRD Research Career Scientist Award Application

Wm S. Middleton Memorial Veterans Hosp, Madison WI

Investigators

Abstract

Ongoing research in my laboratory is focused on three broad areas: 1) understanding the molecular mechanisms that drive melanoma tumor development and progression, 2) defining the role of skin microenvironment on melanomagenesis, and 3) identifying prognostic biomarkers for patients diagnosed with early-stage melanoma. The goal of the research project #1, which is funded by VA BLR&D Merit Review Award, is to understand the role of EPACs, proteins that mediate the alternative cAMP signaling, in promoting the growth of primary melanoma and the mechanism involved in metabolic adaptation that abolishes EPAC dependency during tumor progression. We are exploring inhibition of EPAC signaling in primary melanoma and restoration of EPAC dependency in metastatic melanoma as strategies, respectively, for prevention and treatment of melanoma in Veterans as well as general population. In project #2, we are modeling melanomagenesis using human skin-on-a-chip. The rationale for this project is that while genetically modified mouse models and human melanoma cell lines models in vitro are useful, they do not fully mimic the complex interactions that occur during melanomagenesis in the intact human skin microenvironment. The goal of the DoD Peer Reviewed Medical Research Program-funded research is to understand the role of epidermal keratinocytes and dermal fibroblasts in melanomagenesis to devise strategies for melanoma prevention in active service members and Veterans with increased risk of melanoma. These research projects with in-depth focus on cell and molecular aspects of melanoma also involve translational research using retrospective analysis of fixed and paraffin embedded human primary melanoma tumor tissues. The major focus of my future research is to translate our findings in prospective investigations in Veterans diagnosed with early-stage cutaneous melanoma. Recently, a collaboration with Drs. Gunasekaran and Jose Ayuso, biomedical engineers with expertise in biosensors and microfluidic technology, respectively, allowed us to develop a sensitive biosensor that we propose to employ for prospective studies targeted to detection of circulating melanoma cells in Veteran diagnosed with early-stage melanoma. In proof-of-principle studies, we showed selective and sensitive detection of cells in patient blood. We show that this immunosensor is readily adaptable, in an arrayed format, for simultaneous detection of multiple biomarkers and can be incorporated into a microfluidic device and multiplexed to identify and capture subsets of CTC based on their cell surface markers for phenotypic and molecular characterization. The goal of project #3 is to identify and characterize circulating tumor cells (CTC) in the peripheral blood as indicators of risk of metastatic melanoma and residual disease. The proposed specific aims of this project are a) detection and capture of melanoma cells based on surface marker heterogeneity, b) detection and characterization of CTC heterogeneity using multiplexed microfluidic immuno-sensor array and c) phenotypic and molecular characterization of CTC. These studies will be supported by VA CSR&D Merit Review Award application selected for funding. During the Research Career Scientist Award period, I plan to integrate our understanding of molecular mechanisms in melanoma progression with prospective studies that often require long-term monitoring of the patients. I plan to leverage the microfluidic platform to develop human skin-on-chip to investigate the relationship between environmental factors and risk of melanoma in Veterans. More importantly, the Research Career Scientist Award support will allow us to collect the critical additional data to support my next round of Merit Review application by going beyond detection of circulating melanoma cells to leverage the liquid biopsy for monitoring risk of metastatic melanoma in Veterans.