Intrinsic epithelial defects and NF-κB-dependent signaling heighten viral injury in biliary atresia

Targeted injury to the biliary epithelium and impaired bile flow are central mechanisms underlying childhood cholangiopathies. In biliary atresia (BA), immune-mediated cholangiocyte injury drives progressive bile duct obstruction, yet epithelial-intrinsic programs regulating injury susceptibility remain incompletely defined. Here, we modeled virus-epithelium interactions using rhesus rotavirus (RRV) infection of human liver-derived cholangiocyte organoids from normal donors (NCO) and infants with BA at diagnosis (BACO-Dx) or transplantation (BACO-Tx). Compared with NCO, BA organoids displayed baseline defects in tight junction architecture and barrier function, with prominent epithelial disruption and apoptosis following RRV exposure. Transcriptomic profiling revealed that while NCO and BACO-Dx mounted predominantly interferon-associated antiviral responses, BACO-Tx exhibited an expanded inflammatory program characterized by enrichment of NF-κB (RELA/NFKB1) regulatory network at baseline and amplified by infection. Inhibition of NF-κB signaling with the hydrogen sulfide donor GYY4137 restored organoid growth, improved tight junction organization, reduced permeability, and attenuated RRV-induced apoptosis. In the murine BA model, GYY4137 improved disease severity, preserved bile duct architecture, and protected epithelial junctional integrity while limiting cholangiocyte apoptosis. These findings identify an epithelial NF-κB-predominant program as a key determinant of intrinsic susceptibility to viral injury in BA and support targeting cholangiocyte NF-κB signaling to suppress disease progression.