Lipotoxic fingerprints in clinically relevant postoperative pancreatic fistula: fatty acid driven cytotoxicity targets cells involved in anastomotic healing

Abstract

Background & AimsClinically relevant postoperative pancreatic fistula (CR-POPF) remains a major cause of morbidity following pancreatic surgery, potentially due to fatty acid release by lipase activity. This study investigated how the biochemical composition of CR-POPF effluents drives cellular injury and transcriptional stress responses. MethodsDrain effluents from 14 patients undergoing pancreatoduodenectomy (7 with CR-POPF and 7 controls) were analyzed using gas chromatography-mass spectrometry. Candidate lipids were tested on human foreskin fibroblasts, mesothelial cells, and pancreatic epithelial cells using viability and cytotoxicity assays. Effluents were applied directly to cultures, and RNA sequencing was performed on cells exposed to the two most cytotoxic CR-POPF samples. ResultsMetabolomic profiling revealed lipolytic traits characterized by long-chain saturated fatty acids, including palmitic and stearic acid, and the palmitic acid monoacylglycerol monopalmitin, in drain effluents. These fatty acids accounted for over 70% of the variance in multivariate metabolomic analyses between CR-POPF and control groups. Dose-response assays confirmed concentration-dependent cytotoxicity (p 19, p View larger version (15K): org.highwire.dtl.DTLVardef@2ba175org.highwire.dtl.DTLVardef@752522org.highwire.dtl.DTLVardef@d8f823org.highwire.dtl.DTLVardef@8acb7f_HPS_FORMAT_FIGEXP M_FIG Graphical abstract C_FIG