Grant: Doctoral: Vanier Canada Graduate Scholarships

Pancreatic ductal adenocarcinoma (PDAC) is a deadly malignancy with a low 5-year survival rate and few therapeutic treatments. Malignant cancer cells grow and metabolize rapidly with a high demand for nutrients including amino acids. Alanine (an amino acid) has been shown to act as a significant carbon and nitrogen source for amino acid metabolism, protein and fat synthesis, and energy pathways in pancreatic cancer. PDAC has access to alanine from different sources, such as environmental uptake or from synthesis de novo, which creates a major barrier to understanding why the cells need alanine. My overall objective is to understand why and how alanine restriction impacts PDAC metabolism and growth. Two specific objectives are: (1) to develop an enzymatic depletion strategy to limit alanine in PDAC cells since they can fulfill their demands through de novo synthesis and uptake and (2) to track how the depletion of alanine causes metabolic and cellular stress in PDAC cells. I have successfully generated a stable human pancreatic cancer cell line that expresses the engineered enzyme system and depleted alanine using lentiviral transduction, which is a method that allows the delivery of foreign gene to mammalian cells. Using PDAC cell lines, I will determine how alanine limitation activates cellular stress responses and affects intracellular metabolites by molecular and analytical techniques such as western blot and mass spectrometry. Cell proliferation, survival assay, and tumour growth in mice will also be measured in response to alanine limitation. The expected results will define the direct and indirect metabolic roles for alanine in PDAC cells. In addition, the results of these findings will potentially inform therapeutic development (e.g., targeted metabolic restriction using pharmacologic or enzymatic approaches) for PDAC.