Targeting Treg–fibroblast interaction to enhance immunotherapy in steatotic liver disease-related hepatocellular carcinoma
Aldo Prawira, Hang Xu, Yu Mei, Wei Qiang Leow, N. Jannah M. Nasir, Marie J. Y. Reolo, Masayuki Otsuka, Mohammad Rahbari, Ziyao Chen, Madhushanee Weerasooriya, Liyana Bte Abdullah, Jiawei Wu, Sharifah Nur Hazirah, Martin Wasser, Alexander Yaw Fui Chung, Brian K. P. Goh, Pierce K. H. Chow, Salvatore Albani, Joycelyn Jie Xin Lee, Tony Kiat Hon Lim, Weiwei Zhai, Yock Young Dan, George Boon‐Bee Goh, Mathias Heikenwälder, Yongliang Zhang, Ramanuj DasGupta, Wai Meng David Tai, Haiyan Liu, Jinmiao Chen, Valerie Chew
Abstract
Background Steatotic liver disease-related hepatocellular carcinoma (SLD-HCC), a rising global challenge, is characterised by unique tumour microenvironment (TME) adaptations. Objective This study investigates the immune microenvironment and interactions driving immunosuppression and potential resistance to immunotherapy in SLD-HCC. Design We employed single-cell transcriptomics, cytometry by time-of-flight (CyTOF) and two independent spatial transcriptomics platforms to study the TME of 22 SLD-HCC and 31 non-SLD-HCC cases. Findings were further validated using multiplex immunohistochemistry in an independent cohort of 103 patients, an HCC model and an immunotherapy-treated patient cohort to evaluate clinical relevance. Results Our findings revealed significant alterations in immune and lipid metabolism pathways, particularly in regulatory T cells (Tregs) and cancer-associated fibroblasts (CAFs), suggesting distinct cellular adaptations to a high-fat TME and general immunosuppression. CyTOF revealed a cold, immunosuppressive TME with reduced CD8 + T cells and increased Tregs. Spatial transcriptomics further highlighted distinct Treg–CAF clusters localised at tumour margins, suggesting a spatially organised immunosuppressive niche. Mechanistically, tumour necrosis factor superfamily member 14 (TNFSF14)-tumour necrosis factor receptor superfamily member 14 (TNFRSF14)-mediated Treg–CAF interaction was identified as a critical driver of immunotherapy resistance in SLD-HCC. Blocking TNFRSF14 in an HCC model fed with a high-fat diet resulted in reduced Tregs, increased active CD8 + and memory CD4 + T cells, and a synergistic effect with anti-programmed cell death protein 1 therapy to enhance antitumour immunity and overcome immunotherapy resistance in SLD-HCC. Conclusion This study uncovers critical immune and metabolic adaptations in SLD-HCC, identifying TNFSF14-TNFRSF14 signalling as a key driver of immunotherapy resistance. Targeting this signalling axis enhances antitumour immunity and improves immunotherapy efficacy, offering a promising therapeutic strategy for SLD-HCC.