Precision proteomics of pancreatic ductal adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive cancer and the fifth leading cause of cancer-related death in Western societies. Unlike other major cancers, its survival rate has not improved over the past 40 years, especially among the elderly. PDAC's aggressiveness is due to late detection, rapid metastasis, and resistance to treatment, with current diagnostic methods often missing early stages. A critical challenge is the absence of specific biomarkers to identify PDAC, underscoring the need for new prognostic, diagnostic, and therapeutic biomarkers. PDAC is characterized by hypoxia and limited external nutrients, driving tumor cells to adopt alternative metabolic pathways, including autophagy. Autophagy and the biogenesis of extracellular vesicles (EVs) share many pathways, suggesting that EVs from hypoxic PDAC cells may have distinct molecular profiles, potentially useful for diagnostics. This research aims to develop a method to capture and analyze EVs from PDAC cells, particularly under hypoxic conditions, using Bioorthogonal noncanonical amino acid tagging (BONCAT). BONCAT introduces non-canonical amino acids with an azide side chain into proteins of hypoxic cells, enabling the isolation of these proteins from EVs. The project integrates expertise in EV isolation, high-sensitivity proteomics, genetic engineering to create PDAC cells expressing a mutant methionyl tRNA synthetase under hypoxic conditions, and advanced cell culture techniques.