Abstract 1420: MITF regulates proliferative subpopulation tumor architecture and modifies invasion and characteristics of the epithelial to mesenchymal transition within melanoma

Abstract

Melanoma is the leading cause of skin cancer-related death. Survival rates are high if the disease is diagnosed early, but drop precipitously at later stages. Small molecule inhibitor therapy has given robust responses in the clinic, but relapse is almost certain. This relapse may be due, in part, to tumor heterogeneity. Not only are there multiple cell types within a tumor, but cancer cells themselves can exhibit various phenotypes. This can be due to genotype variation or nutrient availability, and result in populations with different proliferative and invasive capabilities. As these cells display various behaviors, they could also respond to therapies uniquely. Understanding the molecular signature influencing different sub-populations is therefore crucial to design the most effective therapeutic regimen. The fluorescence ubiquitination cell cycle indicator (FUCCI) system, which delineates phases of the cell cycle by visual means, was employed to better understand melanoma tumor heterogeneity. Using this model, it was found that tumor xenografts grown in mice produce two cohorts. One that contained distinct clusters of either arrested or proliferating cells, and another that displayed a homogenous dispersion of proliferating cells throughout the breadth of the tumor. These cohorts were subsequently discovered to display either low or high levels of microphthalmia-associated transcription factor (MITF) expression, respectively. Additionally, loss of MITF by shRNA treatment resulted in conversion of the ability of melanoma cells to give rise to a homogenous xenograft, to instead produce a clustered tumor phenotype. Furthermore, in a 3D in vitro tumor spheroid model, MITF expression was predominantly found in the periphery of the spheroid, which corresponds with the region of highly proliferative cells. Forced over-expression of MITF within these spheroids results in loss of the distinct proliferative ring, and instead a homogenous growth pattern. Not only do spheroids express MITF around the perimeter, but also markers of the Epithelial to Mesenchymal Transition (EMT). These markers, such as Vimentin and Slug, also switch to become expressed homogenously upon high MITF expression. Surprisingly, the increased levels of EMT marker expression by MITF do not correlate to increased migration, and these spheroids in fact show reduced invasion into collagen. We are currently exploring what other means of cancer cell migration could trump an enhanced EMT phenotype and slow invasion in our model. These data outline how tumor heterogeneity, including proliferative and invasive potential, is tightly intertwined with MITF expression, making it an important marker for therapy design. Citation Format: Crystal A. Tonnessen, Kimberley A. Beaumont, David S. Hill, Sheena M. Daignault, Andrea Anfosso, Russell J. Jurek, Wolfgang Weninger, Nikolas K. Haass. MITF regulates proliferative subpopulation tumor architecture and modifies invasion and characteristics of the epithelial to mesenchymal transition within melanoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1420. doi:10.1158/1538-7445.AM2015-1420