Targeting the N-acetyltransferase 10/DKK2 axis enhances CD8+ T cell antitumor activity in colorectal cancer models
Mengmeng Li, Xiaoya Zhao, Jun Wu, Shimeng Zhou, Yao Fu, Chen Chen, Zhuang Ma, Jiawen Xu, Yun Qian, Zhangding Wang, Bo Wang, Qiang Wang, Qingqing Ding, Changyu Chen, Honggang Wang, Xiaozhong Yang, Weijie Dai, Wenjie Zhang, Shouyu Wang
Mengmeng Li, Xiaoya Zhao, Jun Wu, Shimeng Zhou, Yao Fu, Chen Chen, Zhuang Ma, Jiawen Xu, Yun Qian, Zhangding Wang, Bo Wang, Qiang Wang, Qingqing Ding, Changyu Chen, Honggang Wang, Xiaozhong Yang, Weijie Dai, Wenjie Zhang, Shouyu Wang
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Research Article Gastroenterology Immunology Oncology

Targeting the N-acetyltransferase 10/DKK2 axis enhances CD8+ T cell antitumor activity in colorectal cancer models

Abstract

Despite overexpression of N-acetyltransferase 10 (NAT10) in colorectal cancer (CRC), its immunomodulatory role in the tumor microenvironment remains elusive. Here, we reveal that NAT10 promotes immune evasion through N4-acetylcytosine–dependent (ac4C-dependent) mRNA stabilization. Using syngeneic mouse models (MC38/CT-26), intestinal epithelial-cell specific Nat10 conditional KO (Nat10cKO) mice, patient-derived organoids, and clinical specimens, we show that Nat10 ablation enhanced CD8+ T cell–mediated antitumor immunity. Single-cell RNA-seq revealed increased cytotoxic CD8+ T cell infiltration in Nat10cKO tumors, which was corroborated by the inverse correlation of tumoral NAT10 expression and CD8+ T cell number in clinical specimens. Multi-omics integration analysis identified DKK2 as the predominant NAT10-regulated transcript. NAT10 stabilized DKK2 mRNA via ac4C modification, leading to high expression of the DKK2 protein. Secreted DKK2 engaged LRP6 receptors to activate AKT-mTOR signaling, inducing cholesterol accumulation in CD8+ T cells and impairing their cytotoxicity. Pharmacological NAT10 inhibition (Remodelin treatment) or DKK2 neutralization restored CD8+ T cell function and synergized with anti–PD-1 therapy. Our findings establish the NAT10/DKK2/LRP6/AKT-mTOR/cholesterol axis as a critical regulator of CD8+ T cell dysfunction in CRC, positioning NAT10/DKK2 as a potential target to enhance immunotherapy efficacy.

Authors

Mengmeng Li, Xiaoya Zhao, Jun Wu, Shimeng Zhou, Yao Fu, Chen Chen, Zhuang Ma, Jiawen Xu, Yun Qian, Zhangding Wang, Bo Wang, Qiang Wang, Qingqing Ding, Changyu Chen, Honggang Wang, Xiaozhong Yang, Weijie Dai, Wenjie Zhang, Shouyu Wang

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Figure 1

NAT10 deficiency attenuates CRC progression and activates antitumor immunity in syngeneic allografts.

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NAT10 deficiency attenuates CRC progression and activates antitumor imm...
(A) Western blot confirming Nat10 KO in MC38 cells. (B) Schematic for the subcutaneous implantation of WT or Nat10-KO MC38 cells into C57BL/6 mice. (C–E), Tumor growth curves (mean ± SEM) (C), representative images of tumors from each group (D), and tumor weights (E) (n = 7 mice/group). (F) The composition of immune cells in tumors from the MC38 WT and Nat10-KO groups was determined via flow cytometry (n = 5 mice/group). The data are presented as the mean ± SD of indicated mice per group and are representative of 2 independent experiments (C–F). (G) Representative mIHC staining of Nat10, GzmB, CD4+ T cells, CD8+ T cells and macrophages in tumor sections (n = 5 mice/group). Scale bar: 50 μm. (H) Quantification of CD4+ T cell and CD8+ T cell densities in tumor sections (n = 5 mice/group). (I) Flow cytometric analysis of GzmB+ and IFN-γ+CD8+ T cell infiltration in tumors from the MC38 Nat10-WT and KO groups (n = 5 mice/group). (J and K) mIHC-based quantification of exhausted PD-1+ Tim-3+ CD8+ T cells. Representative images are shown in J (scale bar: 50 μm), and quantification analysis is shown in K (n = 3 mice/group). (L) Flow cytometry assessment of Tebt+ and IFN-γ+CD4+ T cell populations. Data are shown as the mean ± SD of indicated mice per group (H, I, K, and L). Statistical analysis was performed by 1-way ANOVA (E, F, H, I, K, and L) and 2-way ANOVA (C). ns, P ≥ 0.05. P < 0.05 was considered to indicate statistical significance.