Pharmacological activation of STAT1-GSDME pyroptotic circuitry reinforces epigenetic immunotherapy for hepatocellular carcinoma

Tu, Yalin
Wu, Haoran
Zhong, Chengpeng
Liu, Yan
Xiong, Zhewen
Chen, Siyun
Wang, Jing
Wong, Patrick Pak-Chun
Yang, Weiqin
Liang, Zhixian
Lu, Jiahuan
Chen, Shufen
Zhang, Lingyun
Feng, Yu
Si-Tou, Willis Wai-Yiu
Yin, Baoyi
Lin, Yingnan
Liang, Jianxin
Liang, Liying
Vong, Joaquim S L
Ren, Weida
Kwong, Tsz Tung
Leung, Howard
To, Ka Fai
Ma, Stephanie
Tong, Man
Sun, Hanyong
Xia, Qiang
Zhou, Jingying
Kerr, David
La Thangue, Nick
Sung, Joseph J Y
Chan, Stephen Lam
Cheng, Alfred Sze-Lok

¹School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong, China,
²Department of Liver Surgery, Renji Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai, China,
³Department of Clinical Pharmacy, Guangzhou Medical University, Guangzhou, China,
⁴Department of Clinical Oncology, The Chinese University of Hong Kong, Hong Kong, China,
⁵Department of Anatomical and Cellular Pathology, The Chinese University of Hong Kong, Hong Kong, China,
⁶School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China,
⁷Nuffield Division of Clinical and Laboratory Sciences, University of Oxford, Oxford, UK,
⁸Department of Oncology, The University of Oxford, Oxford, UK,
⁹Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore,
¹⁰State Key Laboratory of Digestive Disease, The Chinese University of Hong Kong, Hong Kong, China,

Correspondence to Professor Alfred Sze-Lok Cheng; e-mail: [email protected]; Professor Stephen Lam Chan; e-mail: [email protected]

Received February 24, 2024; Accepted October 02, 2024

X @stephaniema912, @caroltong92

YT and HW contributed equally.

Contributors Study concept and design: YT, SLC and AS-LC; Data acquisition and analysis: YT, HW, CZ, YanL, ZX, SiyunC, JW, PP-CW, WY, ZL, JLu, ShufenC, LZ, YF, WW-YS-T, BY, YingnanL, JLiang, LL, JSLV, WR, TTK, HL, JZ, HS, SLC and AS-LC; Clinical resources: SLC and K-FT; Bioinformatics analysis: HW; Writing of manuscript: YT and AS-LC; Material support: SM, DK and NLT; Critical review of manuscript: JZ and AS-LC; Supervision: MT, QX, JJYS, SLC and AS-LC; Funding acquisition: SM, DK, NLT, JJYS, SLC and AS-C. AS-LC is the guarantor.

Funding This project is supported by the General Research Fund (14119023, 14120621 and 14115820), Theme-based Research Scheme (T11-706/18-N), Research Impact Fund (R7022-20), the Li Ka Shing Foundation (CUHK_2021), the CUHK Strategic Seed Funding for Collaborative Research Scheme (NL/SSFCRS2022/0672/22en) and the NSFC/RGC Joint Research Scheme 2023/24 (N_CUHK456/23), We also acknowledge support (funding and study medications) by IngenOx Therapeutics (CUHK_2019) and Merck Sharp and Dohme (MSD-IIS 55253) for the preclinical and clinical studies, respectively.,

Competing interests The authors declare no conflicts of interest related to this work, except for the following declarations. SLC serves as an advisory member for AstraZeneca, MSD, Eisai, BMS, Ipsen and Hengrui, received research funds from MSD, Eisai, Ipsen, SIRTEX and Zailab, and honoraria from AstraZeneca, Eisai, Roche, Ipsen and MSD.

Patient and public involvement Patients and/or the public were not involved in the design, or conduct, or reporting, or dissemination plans of this research.

Provenance and peer review Not commissioned; externally peer reviewed.

Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.

Gut Publish Ahead of Print, November 5, 2024. | DOI: 10.1136/gutjnl-2024-332281

Background

Genomic screening uncovered interferon-gamma (IFNγ) pathway defects in tumours refractory to immune checkpoint blockade (ICB). However, its non-mutational regulation and reversibility for therapeutic development remain less understood.

Objective

We aimed to identify ICB resistance-associated druggable histone deacetylases (HDACs) and develop a readily translatable combination approach for patients with hepatocellular carcinoma (HCC).

Design

We correlated the prognostic outcomes of HCC patients from a pembrolizumab trial (NCT03419481) with tumourous cell expressions of all HDAC isoforms by single-cell RNA sequencing. We investigated the therapeutic efficacy and mechanism of action of selective HDAC inhibition in 4 ICB-resistant orthotopic and spontaneous models using immune profiling, single-cell multiomics and chromatin immunoprecipitation-sequencing and verified by genetic modulations and co-culture systems.

Results

HCC patients showing higher HDAC1/2/3 expressions exhibited deficient IFNγ signalling and poorer survival on ICB therapy. Transient treatment of a selective class-I HDAC inhibitor CXD101 resensitised HDAC1/2/3^high tumours to ICB therapies, resulting in CD8⁺T cell-dependent antitumour and memory T cell responses. Mechanistically, CXD101 synergised with ICB to stimulate STAT1-driven antitumour immunity through enhanced chromatin accessibility and H3K27 hyperacetylation of IFNγ-responsive genes. Intratumoural recruitment of IFNγ⁺GZMB⁺cytotoxic lymphocytes further promoted cleavage of CXD101-induced Gasdermin E (GSDME) to trigger pyroptosis in a STAT1-dependent manner. Notably, deletion of GSDME mimicked STAT1 knockout in abolishing the antitumour efficacy and survival benefit of CXD101-ICB combination therapy by thwarting both pyroptotic and IFNγ responses.

Conclusion

Our immunoepigenetic strategy harnesses IFNγ-mediated network to augment the cancer-immunity cycle, revealing a self-reinforcing STAT1-GSDME pyroptotic circuitry as the mechanistic basis for an ongoing phase-II trial to tackle ICB resistance (NCT05873244).