Immunological Synapse Restricted Metabolic Reprogramming Drives Driectional Cytokine Synthesis

免疫突触限制代谢重编程驱动定向细胞因子合成

• 批准号: 10156024
• 负责人: Ronal Peralta
• 金额: $ 4.6万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10156024
• 关键词: Immunological; Synapse; Restricted; Metabolic; Reprogramming

Project Summary/Abstract T cells become activated when they encounter an antigen presenting cell and receive three signals: TCR signaling via peptide-MHC, co-stimulatory signaling and cytokines. Activation drives a host of metabolically demanding processes such as proliferation, differentiation, migration, and effector functions. Thus, T cells switch from oxidative phosphorylation (OXPHOS), when naïve, to aerobic glycolysis upon activation to generate enough ATP to accommodate these processes. We recently showed that T cells engage aerobic glycolysis within minutes of TCR signaling, independent of co-stimulatory signaling. We found that this mechanism was mediated by pyruvate dehydrogenase kinase 1 (PDHK1), a mitochondrial enzyme that associated with LCK and migrated to the T cell synapse upon activation. These data suggested that aerobic glycolysis could be spatially regulated at the T cell synapse during activation. Using pH sensitive fluorescent systems, we were able to generate data visualizing aerobic glycolysis restricted to the T cell synapse. These data suggest that the mitochondrial positioning in T cells could regulate the initiation of aerobic glycolysis at the immunological synapse. Further, our lab and others have shown that glycolytic enzymes, GAPDH and LDH, are mRNA binding proteins and repress their cytokine translation in naïve T cells, and activation of glycolysis via TCR stimulus promotes the dissociation of glycolytic enzymes from cytokines mRNA. Therefore, we hypothesize that mitochondrial migration to the IS, regulated by mitochondrial LCK, enables localized aerobic glycolysis and subsequent synapse restricted cytokine translation for directed effector functions. To address this hypothesis we will, (1) Identify the role of mitochondrial LCK in driving mitochondrial migration to the T cell synapse and the initiation of aerobic glycolis. Using pH sensitive fluorescence systems, and mitochondrial and LCK reporters, we will visualize the dynamics of aerobic glycolysis, mitochondria and LCK at the T cell synapse when TCR is stimulated by an APC. Moreover, we will (2) Investigate the role of site restricted aerobic glycolysis in promoting localized cytokine translation at the T cell synapse. Using modulators of glycolytic metabolism, we will determine whether aerobic glycolysis could enhance the production of effector cytokines in activated T cells, and whether aerobic glycolysis predicts sites where cytokines are being translated. By better understanding the early signals that promote metabolic reprogramming and drive effector functions we can develop therapeutic targets that will allow us to metabolically modulate T cell activity.

项目摘要/摘要 T细胞在遇到抗原提呈细胞时被激活，并接收三个信号：TCR 通过多肽-MHC、共刺激信号和细胞因子传递信号。激活驱动了大量新陈代谢 要求苛刻的过程，如增殖、分化、迁移和效应器功能。因此，T细胞交换 从天真时的氧化磷酸化(OXPHOS)，到激活后的有氧糖酵解，以产生足够的 ATP来适应这些过程。我们最近发现，T细胞在体内进行有氧糖酵解 几分钟的TCR信号，独立于共刺激信号。我们发现，这一机制是由 通过丙酮酸脱氢酶激酶1(PDHK1)，一种与LCK相关并迁移的线粒体酶 激活后与T细胞突触连接。这些数据表明，有氧糖酵解可以在空间上受到调节。 在激活过程中的T细胞突触。使用pH敏感的荧光系统，我们能够产生数据 可视化有氧糖酵解仅限于T细胞突触。这些数据表明，线粒体 T细胞定位可调节免疫突触有氧糖酵解的启动。此外，我们的 Lab和其他人已经证明糖酵解酶，GAPDH和LDH，是mrna结合蛋白和抑制 它们的细胞因子在幼稚T细胞中的翻译，以及通过TCR刺激激活糖酵解促进解离 从细胞因子信使核糖酵解酶。因此，我们假设线粒体迁移到 受线粒体LCK调控，可实现局部有氧糖酵解和随后的突触 定向效应器功能的限制性细胞因子翻译。为了解决这一假设，我们将：(1)确定 线粒体LCK在驱动线粒体向T细胞突触迁移和启动有氧运动中的作用 甘草甜酒。使用pH敏感的荧光系统，以及线粒体和LCK记者，我们将可视化 APC刺激TCR时T细胞突触的有氧糖酵解、线粒体和LCK的动态变化。 此外，我们将(2)研究部位限制性有氧酵解在促进局部细胞因子中的作用。 T细胞突触的翻译。利用糖酵解代谢的调节剂，我们将确定有氧运动 糖酵解可促进活化T细胞产生效应细胞因子，以及有氧糖酵解是否 预测细胞因子被翻译的位置。通过更好地理解促进 代谢重新编程和驱动效应器功能我们可以开发治疗靶点，使 美国新陈代谢调节T细胞活性。