Comprehensive and Combined Omics Analysis Reveals Immune-Infiltration-Associated Alternative Splicing and RBP Regulators Contributing to Ischemia-Reperfusion Injury in Liver Transplantation.

Background: Hepatic ischemia-reperfusion injury (HIRI) is a major cause of early allograft dysfunction after liver transplantation, but its post-transcriptional regulation remains poorly understood. This study aimed to characterize alternative splicing (AS) events in HIRI and explore a potential association network involving RNA-binding proteins (RBPs), AS, and immune responses. Methods: Using the GSE151648 dataset (80 paired samples from 40 patients), we identified regulated alternative splicing (RAS) events and analyzed dynamic changes in immune cell populations. An RBP-RAS co-expression network was constructed, and single-cell data (GSE171539, one donor, three time points) were used to explore cell-type-specific expression of key RBPs in a qualitative, hypothesis-generating capacity. RBP expression changes were validated in an independent microarray cohort (GSE12720, 20 paired biopsies) and cross-validated with Human Protein Atlas liver proteomic data. Results: Mild IRI (IRI-) samples mainly exhibited alternative 5 Conclusions: This study suggests that mild and severe HIRI are associated with distinct splicing programs and that these changes are closely associated with immune microenvironment remodeling, supporting the hypothesis of an RBP-AS-immune regulatory network. These findings, in which the RBP expression changes were independently validated in an external cohort, provide a basis for generating hypotheses regarding post-transcriptional regulation in HIRI.