Targeting N-acetyltransferase 10 reprograms Treg function to enhance anti–PD-1 immunotherapy sensitivity in murine lung tumors
Xuewei Qi, Dan Cui, Xiao Li, Wenfan Fu, Handong Ma, Wenting Guo, Jiabao Wang, Jiaxi Song, Jiaxin Ma, Jing Yang, Haoyu Yu, Yuting Hong, Qian Wang, Susu Luo, Jia Wang, Baixing Wu, Shanshan Li, Hao Cheng, Min Cheng, Bin Li, Kaiming Zhang, Jia Li, Fengyin Li
Xuewei Qi, Dan Cui, Xiao Li, Wenfan Fu, Handong Ma, Wenting Guo, Jiabao Wang, Jiaxi Song, Jiaxin Ma, Jing Yang, Haoyu Yu, Yuting Hong, Qian Wang, Susu Luo, Jia Wang, Baixing Wu, Shanshan Li, Hao Cheng, Min Cheng, Bin Li, Kaiming Zhang, Jia Li, Fengyin Li
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Research Article Cell biology Oncology

Targeting N-acetyltransferase 10 reprograms Treg function to enhance anti–PD-1 immunotherapy sensitivity in murine lung tumors

Abstract

A significant challenge for improving the efficacy of immune checkpoint blockade (ICB) therapy is the concurrent activation of Tregs, which can dampen antitumor responses. Thus, elucidating the mechanisms that maintain Treg-suppressive function within the tumor microenvironment is critical. Here, we identified an accumulation of N-acetyltransferase 10–modified (NAT10-modified) RNA, particularly in NAT10+FOXP3+ Tregs, in human non–small cell lung cancer. Genetic ablation of Nat10 in tumor-infiltrating (TI) Tregs, or pharmacological inhibition of NAT10 expression, significantly reduced the PD-1+ Treg/CD8+ T cell ratio. These effects were accompanied by an increase in IFN-γ+ Tregs, a reduction in the immunosuppressive activity of Tregs, and inhibition of tumor progression. Mechanistically, NAT10 promotes TI Treg infiltration and activation independently of its RNA-modifying function by directly interacting with FOXP3 and chromatin, modulating local 3D chromatin contacts. Combined administration of anti–PD-1 antibodies and a NAT10 inhibitor further impaired TI Treg–mediated immunosuppression, leading to enhanced tumor control. Our findings demonstrate that NAT10 is a key regulator of TI Treg fragility and function and that its inhibition may enhance the therapeutic efficacy of ICB.

Authors

Xuewei Qi, Dan Cui, Xiao Li, Wenfan Fu, Handong Ma, Wenting Guo, Jiabao Wang, Jiaxi Song, Jiaxin Ma, Jing Yang, Haoyu Yu, Yuting Hong, Qian Wang, Susu Luo, Jia Wang, Baixing Wu, Shanshan Li, Hao Cheng, Min Cheng, Bin Li, Kaiming Zhang, Jia Li, Fengyin Li

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Figure 6

NAT10 promotes iTreg differentiation by positively transcriptional regulation of activation-associated genes.

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NAT10 promotes iTreg differentiation by positively transcriptional regul...
(A) Size-exclusion chromatography analysis of mouse NAT10. (B) EMSA analysis of WT NAT10 binding to DNA. (C) Left: Three-dimensional structure of dimerized NAT10 predicted by AlphaFold3. Two monomers of NAT10 are shown in blue and orange. Right: Surface electrostatic potential of the NAT10 dimer. The positively charged residues and acetyl-CoA-binding residues are colored in pink and green, respectively. (D) EMSA analysis of mutant NAT10 (exon 4–5 deletion) binding to DNA. (E) Sequence conservation analysis. The sequence logo, which depicts the relative frequency of amino acids at each position in a multiple sequence alignment, includes color-coded residues based on their chemical properties. (F) Compared with the WT NAT10 protein, the EMSA analysis of mutant NAT10 (H537A/Y538A) binding to DNA. The binding rate was determined by calculating the proportion of remaining free DNA, as measured by a grayscale value obtained using ImageJ (NIH) (n = 3). (G) The schematic of the experiment. (H–J) Representative histograms (left) and quantification of the MFI (right) of PD-1 (H), 4-1BB (I), and ICOS (J) in iTregs of Nat10+/+ Ox40cre and Nat10fl/fl Ox40cre mice (n = 3). Data represent 2–3 independent experiments; mean ± SEM. P values were calculated using 2-tailed unpaired Student’s t test (F) and 1-way ANOVA (H–J). **P < 0.01, ***P < 0.001, ****P < 0.0001.