ANKRD55 is a key regulator of T cell inflammation in multiple sclerosis
Chuyu Wu, Meiling Jiang, Xue Yang, Yixuan Liu, Bin Huang, Yi Guo, Runjing Cao, Zhihui Cui, Guozhen Deng, Weiyan Wang, Mengdi Guo, Zhiyong Lin, Jiahui Fan, Lin-ming Zhang, Lorenzo Di Cesare Mannelli, Tao Pang, Chenhui Wang, Cun-Jin Zhang
Chuyu Wu, Meiling Jiang, Xue Yang, Yixuan Liu, Bin Huang, Yi Guo, Runjing Cao, Zhihui Cui, Guozhen Deng, Weiyan Wang, Mengdi Guo, Zhiyong Lin, Jiahui Fan, Lin-ming Zhang, Lorenzo Di Cesare Mannelli, Tao Pang, Chenhui Wang, Cun-Jin Zhang
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Research Article Autoimmunity Immunology

ANKRD55 is a key regulator of T cell inflammation in multiple sclerosis

Abstract

Multiple sclerosis (MS) is a progressive, chronic, and highly disabling neuroinflammatory disorder characterized by demyelination and T cell–driven inflammation. Pathogenic T cells play a central role in MS, but effective therapeutic targeting remains challenging. Here, we identified ankyrin repeat domain–containing protein 55 (ANKRD55) as a key regulator of T cell function by single-cell transcriptomic analysis of cerebrospinal fluid and blood from MS patients. ANKRD55 was predominantly expressed in CD4+ T cells in both compartments. Genetic ablation of Ankrd55 led to a robustly reduced disease severity and neuroinflammation in experimental autoimmune encephalomyelitis (EAE), a widely used animal model for MS. Furthermore, T cell–specific deficiency of Ankrd55 significantly impaired Th1 polarization and Th17 differentiation, reducing EAE pathogenicity. Mechanistically, we found that Ankrd55 deficiency disrupted T cell receptor (TCR) signaling integrity. We demonstrated that ANKRD55 regulates the formation of the immune synapse, an essential prerequisite for TCR activation, by interacting with subunits of the chaperonin-containing TCP1 (CCT) complex and modulating its activity, enhancing its assembly by competing with CCT5 for binding to TCP1, CCT3, and CCT6. This facilitates proper microtubule organization and TCR activation. These findings establish ANKRD55 as a critical regulator of TCR signaling and highlight its therapeutic potential in pathogenic T cell–driven autoimmune diseases.

Authors

Chuyu Wu, Meiling Jiang, Xue Yang, Yixuan Liu, Bin Huang, Yi Guo, Runjing Cao, Zhihui Cui, Guozhen Deng, Weiyan Wang, Mengdi Guo, Zhiyong Lin, Jiahui Fan, Lin-ming Zhang, Lorenzo Di Cesare Mannelli, Tao Pang, Chenhui Wang, Cun-Jin Zhang

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Figure 5

ANKRD55 promotes TCR signal transduction.

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ANKRD55 promotes TCR signal transduction. (A) Through KEGG pathway analy...
(A) Through KEGG pathway analysis, the most significantly enriched signaling pathway in CD4+ T cells isolated from the spleens of WT and Ankrd55–/– mice was identified on day 10 after EAE induction. (B) Heatmap analysis of the TCR signaling pathway revealed genes with adjusted P value < 0.05, FDR < 0.05, and log2(fold change) > 1.2 in RNA-Seq data from CD4+ T cells of 3 pairs of WT and Ankrd55–/– mice on day 10 after EAE induction. (C–E) RT-qPCR analysis of CD4+ T cells isolated from the spleens of WT and Ankrd55–/– mice on day 10 after EAE induction (n = 8) demonstrated differential expression of target genes. (F) CD4+ T cells were isolated from the spleens of WT and Ankrd55–/– mice and cultured on plates coated with anti-CD3 and anti-CD28 for 48 hours. IL-2 secretion levels were analyzed using ELISA. (G) Measurement of IL-2 secretion in ANKRD55-overexpressing Jurkat cells. Jurkat cells stably overexpressing ANKRD55 or transfected with empty vector were cultured on plates coated with anti-CD3 and anti-CD28 for 48 hours. Culture supernatants were collected, and IL-2 levels were quantified by ELISA. (H and I) Immunoblot analysis of TCR signaling in primary and transformed T cells. (H) Naive CD4+ T cells isolated from WT and Ankrd55–/– mice were stimulated on plates coated with anti-CD3 and anti-CD28 for 0, 2, 15, or 30 minutes. Cell lysates were collected and subjected to immunoblotting to assess activation of TCR signaling pathways. (I) Jurkat cells stably overexpressing ANKRD55 or transfected with empty vector were treated under the same stimulation conditions, and protein lysates were analyzed by immunoblotting. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, based on unpaired, 2-tailed t test (C–G). Data are shown as mean ± SEM.