Project 2 - Abstract/Summary Our mechanistic work has established general principles of metabolic reprogramming in PDAC, revealing therapeutic insights that show dramatic effects in preclinical models (alanine metabolism blockade) as well as benefit in the clinic (autophagy inhibition). In this next cycle, we seek to address two key questions to enable us to most effectively define and exploit metabolic vulnerabilities in PDAC. First, how do the different concurrent cancer gene mutations that define PDAC subsets influence cancer cell metabolism and the metabolic interplay with the TME? Secondly, what role does regulation of mitochondria, organelles central to integration of metabolism and innate immune signals, play in metabolic reprogramming and cancer-stroma cell interactions in genetic subsets of Kras* PDAC? By developing a series of novel GEM models and patient-derived KRAS* PDAC models representing major genotypes, we have identified pronounced genetically-driven differences in tumor cell metabolism and cancer-immune cell interactions. We hypothesize that cooperating genetic alterations drive distinct metabolic programs and associated vulnerabilities in cancer cells. Moreover, we predict that immune differences will further influence cancer cell metabolism in these distinct genetic settings. To address these hypotheses, we propose the following aims: #1 Determine the influence of major PDAC gene mutations on metabolic programs;? #2 Assess role of the autophagy/lysosome system on tumor-TME crosstalk in genetic subsets of PDAC;? and, #3 Investigate mitochondrial regulation and relationship to metabolism and inflammatory signaling in PDAC subsets.