ACAT1 regulates tertiary lymphoid structures and correlates with immunotherapy response in non–small cell lung cancer
Mengxia Jiao, … , Jie Gu, Ronghua Liu
Mengxia Jiao, … , Jie Gu, Ronghua Liu
Published April 1, 2025
Citation Information: J Clin Invest. 2025;135(7):e181517. https://doi.org/10.1172/JCI181517.
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Research Article Immunology Oncology

ACAT1 regulates tertiary lymphoid structures and correlates with immunotherapy response in non–small cell lung cancer

Abstract

Tertiary lymphoid structures (TLS) in the tumor microenvironment (TME) are emerging solid-tumor indicators of prognosis and response to immunotherapy. Considering that tumorigenesis requires metabolic reprogramming and subsequent TME remodeling, the discovery of TLS metabolic regulators is expected to produce immunotherapeutic targets. To identify such metabolic regulators, we constructed a metabolism-focused sgRNA library and performed an in vivo CRISPR screening in an orthotopic lung tumor mouse model. Combined with The Cancer Genome Atlas database analysis of TLS-related metabolic hub genes, we found that the loss of Acat1 in tumor cells sensitized tumors to anti-PD1 treatment, accompanied by increased TLS in the TME. Mechanistic studies revealed that ACAT1 resulted in mitochondrial protein hypersuccinylation in lung tumor cells and subsequently enhanced mitochondrial oxidative metabolism, which impeded TLS formation. Elimination of ROS by NAC or Acat1 knockdown promoted B cell aggregation and TLS construction. Consistently, data from tissue microassays of 305 patients with lung cancer showed that TLS were more abundant in non–small cell lung cancer (NSCLC) tissues with lower ACAT1 expression. Intratumoral ACAT1 expression was associated with poor immunotherapy outcomes in patients with NSCLC. In conclusion, our results identified ACAT1 as a metabolic regulator of TLS and a promising immunotherapeutic target in NSCLC.

Authors

Mengxia Jiao, Yifan Guo, Hongyu Zhang, Haoyu Wen, Peng Chen, Zhiqiang Wang, Baichao Yu, Kameina Zhuma, Yuchen Zhang, Jingbo Qie, Yun Xing, Pengyuan Zhao, Zihe Pan, Luman Wang, Dan Zhang, Fei Li, Yijiu Ren, Chang Chen, Yiwei Chu, Jie Gu, Ronghua Liu

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Figure 4

Acat1 knockdown promotes the infiltration and activation of B cells in the tumor microenvironment.

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Acat1 knockdown promotes the infiltration and activation of B cells in ...
(A–D) CD45+ immune cells were sorted by flow cytometry from tumor isolated from orthotopic lung tumor mice models. Single-cell sequencing was performed (A), canonical markers of different cell types were used to identify cell clusters (B), and cell clusters are displayed (C). The proportion of different immune cell subsets in the 2 groups (pool of 3 mice per group) (D). (E) The proportion of different immune cell subsets in mouse tumor were verified by flow cytometry, n = 7 per group, experimental replicates = 2. (F–I) Gene-expression levels related to B cell activation (F), proliferation (G), antigen processing and presentation (H), and B-cell receptor signaling pathway of Acat1 negative control (NC) and knockdown (KD) groups. (J–M) Anti-CD20 antibodies were used to eliminate B cells in the KP model, n = 4 per group, experimental replicates = 2. (J), followed by determination of the B cell depletion effect (K), tumor volume (L), tumor weight (M). (N and O) Tertiary lymphoid structures (TLS) in mouse lung tissues of J were observed using H&E and immunofluorescence. Purple, CD8; red, CD19; yellow, CD21; blue, CD23; and grey, DAPI for nucleus. Representative staining images are shown (O). TLS score was determined by the number and maturity of TLS as well as the ratio of the total area of all TLS to the total tumor area, n = 5 per group. (N). Data are shown as the mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 with 2-tailed unpaired Student’s t tests. Please also see Supplemental Figure 3.