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.
View: Text | PDF
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

×

Figure 3

Acat1 knockdown in lung carcinoma cells improves tertiary lymphoid structure abundance and sensitivity to immunotherapy.

Options: View larger image (or click on image) Download as PowerPoint
Acat1 knockdown in lung carcinoma cells improves tertiary lymphoid stru...
(A–C) Tumor development was measured in orthotopic lung tumor mice models. Schematic diagram (A). Acat1 negative control (NC) and knockdown (KD) lung carcinoma cells were inoculated on the lungs of C57/BL6. LLC model, NC and KD n = 10 and 12, respectively, experimental replicates = 3. (B). KP model, n = 4 per group, experimental replicates = 2. (C). (D and E) Tertiary lymphoid structures (TLS) in mice lung tissues of KP model were observed using H&E and immunofluorescence. Purple, CD8; red, CD19; yellow, CD21; blue, CD23; and gray, DAPI for nucleus. Representative staining images are shown (D). TLS score of lung tissues in NC and KD groups were 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 = 14 per group (E). (F) Tumor development for NC and KD groups with or without anti-PD1 treatment was measured in LLC model, NC and KD n = 4 and 6, respectively, experimental replicates = 2. (G and H) Tumor development for NC sg and Acat1 sg groups with or without anti-PD1 treatment was measured in KP-cas9 model, n = 6 per group. (I and J) TLS in G were observed using H&E and immunofluorescence. Purple: CD8; red: CD19; yellow: CD21; blue: CD23; and grey: DAPI. Representative staining images are shown, n = 4 per group. (I). 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 (J). 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 2.