Pancreatic ductal adenocarcinoma (PDAC) is the most common and aggressive cancer of the pancreas. PDAC tumors are highly reliant on nutrient scavenging pathways such as autophagy, and the lysosome to sustain metabolic homeostasis and cellular quality control. To fully understand the functions of the lysosome in PDAC progression, our lab uses biochemical approaches to isolate intact lysosomes from PDAC cells, followed by mass spectrometry-based proteomics. Using this strategy, we have uncovered unique features and functions of PDAC lysosomes that promote cellular adaptation to stress and tumor growth. We have now developed techniques to profile lysosome content and composition during different stages of tumor evolution including metastasis to the liver and lungs – the two major sites of secondary tumor growth in PDAC patients. Our proteomics analysis uncovered pathways and proteins that are unique to lysosomes of tumors growing in these three tissues. For instance, lysosomes isolated from the primary tumor show enrichment for cargo proteins involved in MHC-I mediated antigen processing and presentation, while lysosomes isolated from liver and lung metastases show enrichment for proteins involved in lipoprotein transport and mitophagy, respectively. In addition, we identified hydrolases and transporters that are unique to lysosomes of tumor cells growing in the pancreas, liver or lungs, and their functional inactivation leads to organ specific defects in tumor growth. These studies highlight how dynamic changes in lysosome composition in vivo, enables PDAC cells to rapidly adapt to growth at different organ sites.