Zfp335 establishes eTreg lineage and neonatal immune tolerance by targeting Hadha-mediated fatty acid oxidation
Xin Wang, … , WanJun Chen, Baojun Zhang
Xin Wang, … , WanJun Chen, Baojun Zhang
Published October 16, 2023
Citation Information: J Clin Invest. 2023;133(20):e166628. https://doi.org/10.1172/JCI166628.
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Research Article Autoimmunity Immunology

Zfp335 establishes eTreg lineage and neonatal immune tolerance by targeting Hadha-mediated fatty acid oxidation

Abstract

Regulatory T cells (Tregs) are instrumental in maintaining immune tolerance and preventing destructive autoimmunity, but how heterogeneous Treg populations are established remains largely unknown. Here, we show that Zfp335 deletion in Tregs failed to differentiate into effector Tregs (eTregs) and lose Treg-suppressive function and that KO mice exhibited early-onset lethal autoimmune inflammation with unrestricted activation of conventional T cells. Single-cell RNA-Seq analyses revealed that Zfp335-deficient Tregs lacked a eTreg population and showed dramatic accumulation of a dysfunctional Treg subset. Mechanistically, Zfp335-deficient Tregs displayed reduced oxidative phosphorylation and dysfunctional mitochondrial activity. Further studies revealed that Zfp335 controlled eTreg differentiation by regulating fatty acid oxidation (FAO) through direct targeting of the FAO enzyme Hadha. Importantly, we demonstrate a positive correlation between ZNF335 and HADHA expression in human eTregs. Our findings reveal that Zfp335 controls FAO-driven eTreg differentiation to establish immune tolerance.

Authors

Xin Wang, Lina Sun, Biao Yang, Wenhua Li, Cangang Zhang, Xiaofeng Yang, Yae Sun, Xiaonan Shen, Yang Gao, Bomiao Ju, Yafeng Gao, Dan Liu, Jiapeng Song, Xiaoxuan Jia, Yanhong Su, Anjun Jiao, Haiyan Liu, Lianjun Zhang, Lan He, Lei Lei, WanJun Chen, Baojun Zhang

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Figure 4

Single-cell transcriptomics delineates distinct Treg populations.

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Single-cell transcriptomics delineates distinct Treg populations. (A) Sc...
(A) Schematic diagram of the experimental design, scRNA-Seq, data analysis, and validation. Tregs from 7-day-old WT and KO mice were sorted and subjected to scRNA-Seq by 10x Genomics. (B) Heatmap showing the relative expression of marker genes across different immune cell types. (C) Violin plots showing the Treg marker genes in each Treg cluster. (D) UMAP projections of Treg clusters, color-coded by cluster. (E) Violin plots showing the Treg signature genes specifically expressed in each Treg cluster. (F) Pseudotime plot shows the progression of 4 Treg populations reconstructed by monocle2 using scRNA-Seq data. (G) UMAP projections and percentage of Treg clusters in WT and KO mice. (H) Representative FACS plots of ICOS and CXCR3 expression in splenic Tregs from 8-day-old WT and KO mice. (I and J) Frequencies (I) and numbers (J) of ICOS+CXCR3+ (left) and ICOS+CXCR3– (right) splenic Tregs (n = 3). Gene expression in B, C, and E is represented as the expression of normalized log2 (count +1). Data are representative of 3 independent experiments and are shown as the mean ± SEM (H–J). *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001, by 2-sided, unpaired t tests (I and J).