Prawira Aldo, Xu Hang, Mei Yu, Leow Wei Qiang, Nasir Nurul Jannah Mohamed, Reolo Marie Jy, Otsuka Masayuki, Rahbari Mohammad, Chen Ziyao, Weerasooriya Madhushanee, Abdullah Liyana Bte, Wu Jiawei, Hazirah Sharifah N, Wasser Martin, Chung Alexander, Goh Brian Kp, Chow Pierce Kh, Albani Salvatore, Lee Joycelyn, Lim Tony Kiat Hon, Zhai Weiwei, Dan Yock Young, Goh George Bb, Heikenwälder Mathias F, Zhang Yongliang, Dasgupta Ramanuj, Tai Wai Meng David, Liu Haiyan, Chen Jinmiao, Chew Valerie
Translational Immunology Institute (TII), SingHealth-DukeNUS Academic Medical Centre, Singapore.
Binformatics Institute (BII), Agency for Science, Technology and Research (A*STAR), Singapore.
Gut. 2025 Jul 30. doi: 10.1136/gutjnl-2025-335084.
Steatotic liver disease-related hepatocellular carcinoma (SLD-HCC), a rising global challenge, is characterised by unique tumour microenvironment (TME) adaptations.
This study investigates the immune microenvironment and interactions driving immunosuppression and potential resistance to immunotherapy in SLD-HCC.
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.
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.
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.
脂肪性肝病相关肝细胞癌（SLD-HCC）是一个日益严峻的全球性挑战，其特征在于独特的肿瘤微环境（TME）适应性。
本研究调查SLD-HCC中驱动免疫抑制和免疫治疗潜在抗性的免疫微环境及相互作用。
我们采用单细胞转录组学、飞行时间细胞计数法（CyTOF）以及两个独立的空间转录组学平台，研究22例SLD-HCC和31例非SLD-HCC病例的TME。在一个由103名患者组成的独立队列、一个HCC模型以及一个接受免疫治疗的患者队列中，使用多重免疫组化进一步验证研究结果，以评估临床相关性。
我们的研究结果揭示了免疫和脂质代谢途径的显著改变，特别是在调节性T细胞（Tregs）和癌症相关成纤维细胞（CAFs）中，这表明细胞对高脂肪TME和普遍免疫抑制有独特的适应性。CyTOF显示出一种冷的、免疫抑制性的TME，其中CD8 T细胞减少而Tregs增加。空间转录组学进一步突出了位于肿瘤边缘的不同Treg-CAF簇，表明存在空间组织化的免疫抑制生态位。从机制上讲，肿瘤坏死因子超家族成员14（TNFSF14）-肿瘤坏死因子受体超家族成员14（TNFRSF14）介导的Treg-CAF相互作用被确定为SLD-HCC免疫治疗抗性的关键驱动因素。在喂食高脂肪饮食的HCC模型中阻断TNFRSF14，可导致Tregs减少、活性CD8和记忆CD4 T细胞增加，并与抗程序性细胞死亡蛋白1疗法产生协同效应，以增强抗肿瘤免疫力并克服SLD-HCC中的免疫治疗抗性。
本研究揭示了SLD-HCC中关键的免疫和代谢适应性，确定TNFSF14-TNFRSF14信号传导为免疫治疗抗性的关键驱动因素。靶向该信号轴可增强抗肿瘤免疫力并提高免疫治疗疗效，为SLD-HCC提供了一种有前景的治疗策略。
