Clinical and translational medicine
Graph attention network-driven single-cell omics analysis reveals how B4GALNT1-mediated glycosylation remodelling of the ferroptosis-pyroptosis network promotes drug resistance in hepatocellular carcinoma
By Bie L, Xu B, Shen H and 4 others.
Abstract
Background
Acquired resistance to sorafenib in hepatocellular carcinoma (HCC) is accompanied by coordinated changes in oxidative-stress handling and regulated cell-death pathways, yet how membrane organisation connects these adaptations is not fully resolved. We therefore examined whether B4GALNT1, a ganglioside biosynthetic glycosyltransferase, couples changes in ganglioside composition and lipid-raft architecture to ferroptosis regulation and caspase-1/GSDMD-associated signalling in sorafenib-resistant HCC.
Methods
We combined TCGA-LIHC analysis, single-cell transcriptomics, pathway-level analyses, and graph attention network modelling with a treatment-annotated local cohort of 92 patients with HCC who received sorafenib. Functional experiments were conducted in Huh7/Huh7-SR cells and additional liver-tumour models using B4GALNT1 gain- and loss-of-function, an independent second-sgRNA knockout clone, GM1 rescue, and re-expression of either wild-type B4GALNT1 or the catalytic-loss D358A variant. Validation included ganglioside profiling, detergent-resistant membrane isolation, lipid-raft disruption/repletion, ferroptosis-directed pharmacology, cell-death measurements, and xenograft studies.