Abstract P101: Investigating the Role of Hypoxia-Mediated Metabolic Dysregulation in Immune Evasion in Pancreatic Ductal Adenocarcinoma (PDAC)

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is known for its poor prognosis, with a 5-year survival rate of less than 10%, primarily due to late-stage diagnosis and the absence of reliable early biomarkers. The tumour's poor vascularization and hypoxic microenvironment drive metabolic adaptations that facilitate immune evasion. Despite these critical factors, the interplay between hypoxia-induced metabolic dysregulation and immune regulation in PDAC remains insufficiently explored. This project aims to uncover novel interactions between these processes to identify potential therapeutic targets. PDAC cell lines PANC-1 and BXPC-3, and control non-cancerous pancreatic ductal cells (H6c7) were treated for 72h in hypoxia (1% O2) and RNA was extracted. RNA sequencing was performed in normoxic and hypoxic samples looking for clusters of dysregulated metabolic genes. In total, 59 metabolic genes were commonly upregulated in hypoxic PDAC cells, including; ADAMTS7, PLOD1, COL5A1, DEPTOR, ECM1, GFPT2, HTRA3, ITGB8, and MLLT3. Immune deconvolution was conducted with TIMER database which showed correlation of several of the genes with M2 macrophages, known to promote immune evasion. Overall survival analysis on Kaplan-Meier plotter database also showed correlation of several genes with poor overall survival. 15 genes were selected for further investigation in vitro. siRNA knockdown of the selected genes will be conducted on the PDAC cells, after which conditioned media will be collected and used on THP-1 and U-937 derived macrophages to study effects on macrophage polarisation. For this immunophenotyping with qPCR for macrophage phenotype specific markers will be conducted. This may lead to discovery of novel biomarkers for PDAC diagnosis and targets for therapy.