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 3

ANKRD55 critically promotes Th1 and Th17 differentiation.

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ANKRD55 critically promotes Th1 and Th17 differentiation. (A and B) Flow...
(A and B) Flow cytometry analysis of naive CD4+ T cells sorted by magnetic beads was performed. The cells were cultured on anti-CD3/CD28–coated plates under Th1 (n = 18) or Th17 (n = 12) polarization conditions. (C and D) Naive CD4+ T cells were isolated from WT and Ankrd55–/– mice and cultured under Th1- or Th17-polarizing conditions for 4 days. Expression of T-bet (C) (under Th1 conditions) and RORγt (D) (under Th17 conditions) was evaluated by intracellular staining followed by flow cytometry. Representative flow cytometry plots (left) and quantification of T-bet+ and RORγt+ cells (right) are shown (n = 10 or 12). (E and F) Cells isolated from the spleens of mice on day 10 after immunization were restimulated with MOG35–55 and induced to polarize toward Th1 or Th17, followed by intracellular staining for IL-17A and IFN-γ and analysis by flow cytometry (n = 12). (G) CD4+ T cells were sorted from mouse spleens using magnetic beads. The cells were labeled with CFSE (0.5 μM at 37°C for 5 minutes), and the proportion of proliferating cells was analyzed by flow cytometry after culturing for 48 hours on plates coated with anti-CD3 and anti-CD28 (n = 8). (H) Human PBMCs were divided into different groups based on single nucleotide polymorphism genotype of ANKRD55. Expression of ANKRD55 mRNA was detected by RT-qPCR (n = 11). (I–K) The proportion of CD4+ T cells secreting IFN-γ, IL-17A, and GM-CSF in PBMCs was analyzed by flow cytometry (n = 11). (I and J) IFN-γ and IL-17A; (K) GM-CSF. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, based on unpaired, 2-tailed t test. Data are shown as mean ± SEM.