Integrated Single-Cell and Transcriptomic Analysis Suggests a Cuproptosis-Related Phenotype and Potential Therapeutic...

Acute pancreatitis (AP) is an acute inflammatory disorder of the exocrine pancreas with limited therapies. Whether cuproptosis, a copper-dependent cell death pathway, contributes to AP pathogenesis remains unknown. We integrated single-cell transcriptomics (GSE279876), single-cell transcriptomics (GSE188819), and human whole-blood transcriptomic data (GSE194331) to evaluate 13 cuproptosis-related genes across cell types, along with pseudotime trajectory, regulatory networks, and cell-cell communication. Findings were validated in caerulein-induced severe acute pancreatitis (SAP) mouse models and acinar cell injury using histopathology, electron microscopy, flow cytometry, immunofluorescence, and biochemical assays. Single-cell analysis showed selective activation of a cuproptosis-related signature in acinar cells, with all 13 genes upregulated, most notably GIs and Fdx1. The cuproptosis-related gene signature score increased along with pseudotime, and exploratory transcription factor (TF) and cell-cell communication analyses suggested enrichment of SPI1 and AP-1 (FOS/JUN), alongside MIF/CCL inflammatory signals, which was corroborated in independent datasets. In vitro and in vivo, pancreatic tissues and pancreatic acinar cells in SAP exhibited copper accumulation, FDX1/DLAT upregulation, mitochondrial shrinkage, and cristae loss, which are consistent with a cuproptosis-related phenotype, and were all attenuated by Tetrathiomolybdate (TTM). These findings suggest that the inflammatory microenvironment may be associated with selective activation of a cuproptosis-related program in acinar cells in AP, and that copper chelation with TTM attenuates pancreatic injury, indicating a potential therapeutic strategy targeting copper dyshomeostasis.