Single-cell transcriptome sequencing reveals hepatocyte heterogeneity in response to radiation-induced liver injury and regeneration.

Abstract

Radiation-induced liver injury (RILI) presents a significant constraint on radiotherapy efficacy for hepatic malignancies, yet its underlying mechanisms remain poorly defined. Employing integrated single-cell and single-nucleus RNA sequencing in a rat model receiving 3/4 vol partial liver irradiation (25 Gy), combined with proteomic profiling of human liver tissues, we delineated the spatiotemporal dynamics of hepatocyte responses to RILI. Hepatocyte subpopulations exhibited a compartmentalized division of labor: midzonal (zone 2) hepatocytes were characterized by transient proliferation in conjunction with coagulation factors (Fga/Fgb/Fgg) and ERK signaling, whereas periportal and pericentral (zone 1and 3) hepatocytes were characterized by p53-mediated cell cycle restraint and metabolic adaptation involving PPARα-YAP1-TEAD1 signaling. Trajectory analysis supported zone 2 hepatocytes as the primary source for repopulating damaged zone 1/3 compartments, with periportal regions (zone 1) replenishing first. Notably, pericentral (zone 3) hepatocytes demonstrated heightened vulnerability, manifesting late-stage hypertrophy and S-phase arrest. Interzonal crosstalk, including VEGFA-KDR signaling from zone 1/3 that was associated with zone 2 vascular remodeling and HGF-MET/GAS6-MERTK pathways that may coordinate regeneration and survival, may further contribute to repair. Critically, proteomic profiling of irradiated human liver tissues validated conserved activation of p53, coagulation, ERK, and PPAR/YAP pathways. Our findings unveil a sophisticated spatial paradigm of liver regeneration following radiation injury, wherein spatially partitioned hepatocyte subpopulations deploy compartmentalized repair programs to balance proliferative efficiency and metabolic adaptation, suggesting potential new targets for therapeutic intervention against RILI.