Themicroenvironmentplaysauniqueroleinthedevelopmentofmelanoma,whicharisesintheskin.Theskin displays externally visible signs of aging, such as decreased elasticity and a loss of collagen integrity. Much workhasfocusedrecentlyonhowchangesincollagencanaffecttumormetastasis,bothbiomechanicallyand biochemically. Data from several groups indicate that the cross-talk between stromal fibroblasts and transformed melanocytes is important for invasion, melanoma growth, and even therapy resistance. Our proteomicsanalysesindicatethatfactorsusedtocrosslinkcollagen,suchasHAPLN1aresecretedbyyoung, but not aged dermal fibroblasts, and Wnt5A is secreted by aged, but not young fibroblasts. The interplay between these molecules may contribute to age-related changes in collagen integrity. We hypothesize that changes regulated by age-related alterations in the extracellular matrix (ECM) initiate or promote a pro- metastaticprogram,andimpacttherapyresistance. We will query the contribution of aged skin biochemical and architectural contributions in governing melanoma?smetastaticprogressionaswellasresistancetotargetedtherapy.Weproposeanalternatetheory of matrix stiffness that hypothesizes that increasing stiffness will have a non-linear effect on metastatic progression,andtherapyresistance.Wewillusepathophysiologicallyrelevantinvitro3Dstromalsystems(3D skin reconstructions), animal models of melanoma, a novel simultaneous multi-channel immunofluorescent analysis (SMIA) of formalin-fixed and paraffin embedded (FFPE) human tissue cohorts in combination with a customized new software written for the bulk analysis and acquisition of SMIA-generated images, single moleculeRNAimagingcoupledwithhigh-throughputsinglecellimagingandsequencing,allofwhichwillfeed back into an increasingly intricate mathematical modeling for improved understanding and better patient personalized (i.e., metastatic and efficacy of drug treatment) prediction capabilities. We expect our data will reveal a synergistic picture of the mechanochemical interactions between the aged microenvironment and singulartumorcellsthattheindividualapproachescouldnothavedeciphered. This team project involves experts in the biology of melanoma metastasis, Wnt signaling and aging (Weeraratna), single cell RNA systems biology (Raj), tumor-stromal interactions and digital imaging quantitativeanalyses(Cukierman)andcomputationalandmathematicalpredictivemodeling(Shenoy).
The rate of melanoma development, progression and therapy resistance is greatly increased in the aging population, however the reasons for this are still unclear. We analyzed normal skin cells (fibroblasts) for changes during aging, and found that the most significant changes that occurred were those involved in the architecture of collagen and the integrity of the extracellular matrix. We will use sophisticated mathematical models, high-level imaging studies, and the analysis of the change in the tensile forces of collagen on single cellstouncovermechanismsbywhichthebreakdownofcollagenandtheextracellularmatrixintheagingskin promotesmelanomametastasisandtherapyresistance.