Single-Cell Multiomics Decoding of TCIRG1-Mediated Cuproptosis Circuitry Rewiring Immune-Metabolic Landscape in Ischemic Stroke.

Abstract

Ischemic stroke is one of the leading causes of disability and mortality worldwide. Its pathogenesis extends beyond focal cerebral ischemia-induced neuronal injury, encompassing profound immune dysregulation and secondary inflammatory responses. Recent studies have highlighted T cell dysfunction, particularly T cell exhaustion, as a critical driver of post-stroke immunosuppression and increased susceptibility to infection. However, the molecular mechanisms underlying this process remain unclear. This study focuses on a novel immunoregulatory axis involving cuproptosis, metabolic reprogramming, and T cell exhaustion, and systematically investigates the role of the key regulatory molecule TCIRG1 in post-ischemic immune homeostasis disruption. Through integrative analysis of human peripheral blood transcriptomic data (GSE58294) and single-cell transcriptomic datasets from mouse brain and bone marrow following middle cerebral artery occlusion (MCAO) (GSE174574 and GSE293098), we found that the cuproptosis pathway was markedly activated after ischemic stroke. Notably, TCIRG1 was highly expressed in T cells and co-upregulated with the cuproptosis regulator Fdx1 and T cell exhaustion markers. Metabolic flux analysis revealed that high TCIRG1 expression was associated with enhanced pyruvate metabolism and was accompanied by downregulation of MYC signaling and upregulation of Zfp644, suggesting a role for TCIRG1 in driving T cell exhaustion through metabolic reprogramming. Furthermore, CellChat analysis indicated that TCIRG1 altered intercellular communication between T cells and microglia, thereby reshaping the local immune communication network. Collectively, these findings suggest that TCIRG1 may promote T cell dysfunction via cuproptosis and metabolic pathways, contributing to immune microenvironmental imbalance after ischemic stroke. This study proposes a novel multi-axis regulatory model and provides a potential molecular target for post-stroke immunotherapy.