Metabolic reprogramming by mutant GNAS creates an actionable dependency in intraductal papillary mucinous neoplasms of the pancreas.

[BACKGROUND] Oncogenic 'hotspot' mutations of and are two major driver alterations in intraductal papillary mucinous neoplasms (IPMNs), which are precursors to pancreatic ductal adenocarcinoma. We previously reported that pancreas-specific and co-expression in p48; Kras; Rosa26; Tg (TetO-Gnas) mice (' mice) caused development of cystic lesions recapitulating IPMNs.

[OBJECTIVE] We aim to unveil the consequences of mutant expression on phenotype, transcriptomic profile and genomic dependencies.

[DESIGN] We performed multimodal transcriptional profiling (bulk RNA sequencing, single-cell RNA sequencing and spatial transcriptomics) in the ' autochthonous model and tumour-derived cell lines ( cells), where expression is inducible. A genome-wide CRISPR/9 screen was conducted to identify potential vulnerabilities in co-expressing cells.

[RESULTS] Induction of -and resulting Galpha signalling-leads to the emergence of a gene signature of gastric (pyloric type) metaplasia in pancreatic neoplastic epithelial cells. CRISPR screening identified the synthetic essentiality of glycolysis-related genes and in ; co-expressing cells. Real-time metabolic analyses in cells and autochthonous model confirmed enhanced glycolysis on induction. Induction of made expressing cells more dependent on glycolysis for their survival. Protein kinase A-dependent phosphorylation of the glycolytic intermediate enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) was a driver of increased glycolysis on induction.

[CONCLUSION] Multiple orthogonal approaches demonstrate that and co-expression results in a gene signature of gastric pyloric metaplasia and glycolytic dependency during IPMN pathogenesis. The observed metabolic reprogramming may provide a potential target for therapeutics and interception of IPMNs.