Table_3_The Metabolic Landscape of Thymic T Cell Development In Vivo and In Vitro.xlsx

Although metabolic pathways have been shown to control differentiation and activation in peripheral T cells, metabolic studies on thymic T cell development are still lacking, especially in human tissue. In this study, we use transcriptomics and extracellular flux analyses to investigate the metabolic profiles of primary thymic and in vitro-derived mouse and human thymocytes. Core metabolic pathways, specifically glycolysis and oxidative phosphorylation, undergo dramatic changes between the double-negative (DN), double-positive (DP), and mature single-positive (SP) stages in murine and human thymus. Remarkably, despite the absence of the complex multicellular thymic microenvironment, in vitro murine and human T cell development recapitulated the coordinated decrease in glycolytic and oxidative phosphorylation activity between the DN and DP stages seen in primary thymus. Moreover, by inducing in vitro T cell differentiation from Rag1-/- mouse bone marrow, we show that reduced metabolic activity at the DP stage is independent of TCR rearrangement. Thus, our findings suggest that highly conserved metabolic transitions are critical for thymic T cell development.尽管已有研究证实代谢通路可调控外周T细胞的分化与活化，但针对胸腺T细胞发育的代谢研究仍相对匮乏，尤其是在人体组织样本中。本研究借助转录组学（transcriptomics）与细胞外通量分析（extracellular flux analyses），对原代小鼠及人胸腺细胞、体外诱导分化的小鼠与人类胸腺细胞的代谢谱进行了系统探究。核心代谢通路，特别是糖酵解与氧化磷酸化，在小鼠和人类胸腺的双阴性（DN）、双阳性（DP）以及成熟单阳性（SP）发育阶段间均发生了显著变化。值得关注的是，即便缺乏复杂的多细胞胸腺微环境，体外诱导的小鼠与人类T细胞发育过程，仍重现了原代胸腺中DN阶段向DP阶段过渡时，糖酵解与氧化磷酸化活性协同下降的特征。此外，本研究通过从Rag1-/-小鼠骨髓中体外诱导T细胞分化，证实DP阶段的代谢活性降低并不依赖于T细胞受体（TCR）重排。综上，本研究结果表明，高度保守的代谢转化过程对胸腺T细胞发育至关重要。