| Autor: |
Makino, Yuki, Rajapakshe, Kimal I, Chellakkan Selvanesan, Benson, Okumura, Takashi, Date, Kenjiro, Dutta, Prasanta, Abou-Elkacem, Lotfi, Sagara, Akiko, Min, Jimin, Sans, Marta, Yee, Nathaniel, Siemann, Megan J, Enriquez, Jose, Smith, Paytience, Bhattacharya, Pratip, Kim, Michael, Dede, Merve, Hart, Traver, Maitra, Anirban, Thege, Fredrik Ivar |
| Zdroj: |
Gut; 2025, Vol. 74 Issue: 1 p75-88, 14p |
| Abstrakt: |
BackgroundOncogenic ‘hotspot’ mutations of KRASand GNASare two major driver alterations in intraductal papillary mucinous neoplasms (IPMNs), which are bona fideprecursors to pancreatic ductal adenocarcinoma. We previously reported that pancreas-specific KrasG12Dand GnasR201Cco-expression in p48Cre; KrasLSL-G12D; Rosa26LSL-rtTA; Tg (TetO-GnasR201C) mice (‘Kras;Gnas’ mice) caused development of cystic lesions recapitulating IPMNs.ObjectiveWe aim to unveil the consequences of mutant GnasR201Cexpression on phenotype, transcriptomic profile and genomic dependencies.DesignWe performed multimodal transcriptional profiling (bulk RNA sequencing, single-cell RNA sequencing and spatial transcriptomics) in the ‘Kras;Gnas’ autochthonous model and tumour-derived cell lines (Kras;Gnascells), where GnasR201Cexpression is inducible. A genome-wide CRISPR/Cas9 screen was conducted to identify potential vulnerabilities in KrasG12D;GnasR201Cco-expressing cells.ResultsInduction of GnasR201C—and resulting G(s)alpha 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 Gpi1and Slc2a1in KrasG12D;GnasR201Cco-expressing cells. Real-time metabolic analyses in Kras;Gnascells and autochthonous Kras;Gnasmodel confirmed enhanced glycolysis on GnasR201Cinduction. Induction of GnasR201Cmade KrasG12Dexpressing 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 GnasR201Cinduction.ConclusionMultiple orthogonal approaches demonstrate that KrasG12Dand GnasR201Cco-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. |
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