Integrated Transcriptomic Analyses and Experimental Validation Identify TSPO as a Mitochondria-Associated Therapeutic Target in Myocarditis.

Myocarditis is characterized by inflammatory cell infiltration and myocardial injury, yet current treatments remain largely empirical and nonspecific. Given the heart's high energy demand and the essential role of mitochondria in cellular metabolism, investigating mitochondria-related genes may provide novel therapeutic insights. In this study, we integrated bulk RNA sequencing data from CVB3 viral myocarditis (VMC) model with mitochondrial gene sets from MitoCarta3.0 and human myocarditis-related genes from GeneCards. Through comprehensive bioinformatics analyses, including weighted gene coexpression network analysis (WGCNA), random forest (RF), and SHapley Additive exPlanations (SHAP), we identified mitochondrial metabolism-related genes associated with myocarditis. Single-cell RNA sequencing (scRNA-seq) analysis further revealed that macrophages were the predominant infiltrating immune population, with Tspo expressed in myocarditis macrophages. Tspo knockdown enhanced inflammatory responses and disrupted mitochondrial structure and function, whereas treatment with the Tspo ligand Ro5-4864 exerted the opposite effects. Consistently, administration of Ro5-4864 in a VMC mouse model mitigated inflammation and preserved cardiac function. Collectively, our findings identify Tspo as a novel mitochondria-associated protective response factor in myocarditis, providing new insights into its pathophysiological mechanisms and highlighting Tspo as a promising therapeutic target for future intervention.