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
- 1The Seventh People's Hospital Affiliated to Shanghai University of Traditional Chinese Medicine
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. Results B4GALNT1 was enriched in tumour‐associated hepatocyte populations and was upregulated in Huh7‐SR cells. In the local cohort, higher B4GALNT1 protein abundance tracked with less favourable response to sorafenib and poorer survival outcomes. Increasing B4GALNT1 shifted ganglioside abundance toward GM2/GM1/GD2 and increased GM1‐enriched raft‐associated fractions, whereas knockout produced reciprocal changes. B4GALNT1‐high cells showed increased GPX4 and SLC7A11, reduced lipid peroxidation and labile Fe2+, and lower ferroptotic injury. At the same time, caspase‐1/GSDMD‐associated signalling and IL‐1β/IL‐18 release increased without widespread terminal membrane lysis. Exogenous GM1 partially rescued knockout‐associated changes, and wild‐type B4GALNT1 restored the phenotype more effectively than the D358A mutant. Concordant directional effects were observed in additional cell models and in xenografts. Conclusions The data are consistent with a model in which B4GALNT1 reshapes ganglioside composition and GM1‐enriched membrane microdomains, thereby supporting a drug‐adapted state that combines greater ferroptosis resistance with restricted caspase‐1/GSDMD‐associated membrane permeabilisation. This integrated phenotype accompanies reduced sorafenib responsiveness and offers a mechanistic basis for acquired resistance in HCC. HIGHLIGHTS B4GALNT1 is associated with sorafenib resistance and poorer outcomes in treated HCC. B4GALNT1 remodels gangliosides and lipid rafts, limiting ferroptotic injury. Caspase‐1/GSDMD signalling increases without widespread terminal membrane lysis. Genetic rescue and cross‐model studies support B4GALNT1 specificity.
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BibTeX
@article{bie2026graph,
title = {{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}},
author = {Lihan Bie and Biao Xu and Haitao Shen and Shuwen Ren and Xiaoyang Yao and Zhouhong Xiang and Zheng Xu},
journal = {Clinical and translational medicine},
year = {2026},
month = oct,
doi = {10.1002/ctm2.70873},
url = {https://europepmc.org/article/PMC/PMC13670205},
note = {CC BY},
}
RIS
TY - JOUR
TI - 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
AU - Lihan Bie
AU - Biao Xu
AU - Haitao Shen
AU - Shuwen Ren
AU - Xiaoyang Yao
AU - Zhouhong Xiang
AU - Zheng Xu
PY - 2026
DA - 2026/10/01
JO - Clinical and translational medicine
DO - 10.1002/ctm2.70873
UR - https://europepmc.org/article/PMC/PMC13670205
AB - 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. Results B4GALNT1 was enriched in tumour‐associated hepatocyte populations and was upregulated in Huh7‐SR cells. In the local cohort, higher B4GALNT1 protein abundance tracked with less favourable response to sorafenib and poorer survival outcomes. Increasing B4GALNT1 shifted ganglioside abundance toward GM2/GM1/GD2 and increased GM1‐enriched raft‐associated fractions, whereas knockout produced reciprocal changes. B4GALNT1‐high cells showed increased GPX4 and SLC7A11, reduced lipid peroxidation and labile Fe2+, and lower ferroptotic injury. At the same time, caspase‐1/GSDMD‐associated signalling and IL‐1β/IL‐18 release increased without widespread terminal membrane lysis. Exogenous GM1 partially rescued knockout‐associated changes, and wild‐type B4GALNT1 restored the phenotype more effectively than the D358A mutant. Concordant directional effects were observed in additional cell models and in xenografts. Conclusions The data are consistent with a model in which B4GALNT1 reshapes ganglioside composition and GM1‐enriched membrane microdomains, thereby supporting a drug‐adapted state that combines greater ferroptosis resistance with restricted caspase‐1/GSDMD‐associated membrane permeabilisation. This integrated phenotype accompanies reduced sorafenib responsiveness and offers a mechanistic basis for acquired resistance in HCC. HIGHLIGHTS B4GALNT1 is associated with sorafenib resistance and poorer outcomes in treated HCC. B4GALNT1 remodels gangliosides and lipid rafts, limiting ferroptotic injury. Caspase‐1/GSDMD signalling increases without widespread terminal membrane lysis. Genetic rescue and cross‐model studies support B4GALNT1 specificity.
SN - 2001-1326
ER -