Regulation of encephalitogenic T cells by CRAC channels

CRAC 通道对致脑炎 T 细胞的调节

• 批准号: 10461826
• 负责人: STEFAN FESKE
• 金额: $ 48.93万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-14 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10461826
• 关键词: Adverse effects; Affect; Animal Model; Anti-Inflammatory Agents; Attenuated; Autoimmune; Autoimmune Diseases; Autoimmune Responses; Bacterial Infections; Benefits and Risks; Brain; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CREB1 gene; Calcineurin; Calcium; Calcium Channel; Calcium ion; Cell membrane; Cell physiology; Cells; Cellular Immunity; Complication; Data; Development; Disease; Disease Progression; Dose; Drug Targeting; Enzymes; Experimental Autoimmune Encephalomyelitis; Functional disorder; Gene Expression; Genes; Genetic; Glycolysis; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Helper-Inducer T-Lymphocyte; Homologous Gene; Human; Immune Tolerance; Immunity; Immunotherapy; Impairment; In Vitro; Infection; Inflammation; Inflammatory Response; Interferon Type II; Interleukin-17; Mediating; Mediator of activation protein; Metabolic Pathway; Metabolism; Molecular; Multiple Sclerosis; Multiple Sclerosis Lesions; Mus; Mutagenesis; Mycoses; Myelin; Myelin Proteins; Myelin Sheath; Neuraxis; Neurologic Symptoms; Neurons; Oxidative Phosphorylation; Pathogenicity; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Phosphotransferases; Process; Production; Proteins; Recurrent disease; Regulation; Regulatory T-Lymphocyte; Relapse; Risk; Role; STIM1 gene; Safety; Severities; Severity of illness; Spinal Cord; Symptoms; T-Lymphocyte; T-Lymphocyte Subsets; T-bet protein; Testing; Therapeutic; Virus Diseases; autoimmune inflammation; autoreactive T cell; autoreactivity; brain cell; chronic autoimmune disease; cytokine; efficacy evaluation; in vivo; inhibitor; interleukin-23; mouse model; multiple sclerosis patient; multiple sclerosis treatment; novel strategies; prevent; transcription factor; treatment strategy

Project Summary Multiple Sclerosis (MS) is an autoimmune disorder that is characterized by inflammation of the central nervous system (CNS) and localized destruction of the brain and spinal cord resulting in debilitating neurological symptoms. The disease processes underlying MS have been studied extensively in an animal model called experimental autoimmune encephalomyelitis (EAE), which greatly helped to develop several approved therapies for MS. In both MS and EAE, CD4+ helper T cells mistakenly recognize myelin protein expressed by brain cells as foreign and mount an autoimmune response against it. This inflammatory response is mediated mainly by two subsets of helper T cells, so-called Th1 and Th17 cells, which infiltrate the CNS and produce the proinflammatory cytokines IFNγ and IL-17, respectively. Interfering with the development of naive CD4+ T cells into Th1 and Th17 cells by genetic deletion of the transcription factors T-bet and RORγt, respectively, protects mice from EAE. Our lab showed that CD4+ T cells require the influx of calcium ions for their activation and ability to produce IFNγ and IL-17. Calcium influx is mediated by CRAC (calcium release-activated calcium) channels that are located in the plasma membrane of cells and formed by ORAI1 proteins. ORAI1 is activated by two intracellular proteins, STIM1 and STIM2, and deletion or mutagenesis of either of these three proteins attenuates CRAC channel function and calcium influx. Using mice with genetic deletion of ORAI1, STIM1 and STIM2 as well as specific CRAC channel inhibitors, we found that Th1 and Th17 cells are more dependent on calcium influx than other T cells for their function and ability to cause autoimmune inflammation. Treatment of T cells of mice or humans with a specific CRAC channel inhibitor reduced the production of IL-17 and IFNγ but did not affect T regulatory (Treg) cells. in a dose dependent manner. By genetically ablating either ORAI1, STIM1 or STIM2 in T cells, we could prevent or ameliorate the development of EAE and CNS inflammation in mice. Importantly, deletion of ORAI1 or STIM1 in T cells after EAE symptoms had already developed stopped the progression of disease. Similarly, treatment of mice in which EAE had developed with a CRAC channel inhibitor also significantly reduced disease severity without apparent adverse effects. These data suggest that calcium influx is an essential mediator of CNS inflammation in EAE and that its inhibition may be a new option for the treatment of MS. Drugs inhibiting CRAC channels have been developed, but have not been tested for their efficacy and safety in the treatment of MS and other T cell mediated autoimmune diseases. The goals of this application are twofold: (1) To understand the molecular mechanisms by which calcium influx via CRAC channels controls the development of CD4 T cells into pathogenic T cells, in particular Th1 and Th17 cells, that mediate CNS inflammation and EAE/MS. (2) To evaluate CRAC channels as drug targets for immunotherapy of EAE and eventually MS by studying their role in human T cells, in a mouse model of relapsing-remitting EAE and in immunity to infections as a potential complication of antiinflammatory therapy.

项目概要 多发性硬化症 (MS) 是一种自身免疫性疾病，其特征是中枢神经系统炎症 系统（CNS）以及大脑和脊髓的局部破坏，导致神经衰弱 多发性硬化症的疾病过程主要在称为“多发性硬化症”的动物模型中进行研究。 实验性自身免疫性脑脊髓炎（EAE），这极大地帮助了开发几种已批准的 在 MS 和 EAE 中，CD4+ 辅助 T 细胞错误地识别由 MS 表达的髓磷脂蛋白。 脑细胞被视为异物并对其产生自身免疫反应，这种炎症反应是介导的。 主要由辅助 T 细胞的两个子集（所谓的 Th1 和 Th17 细胞）组成，它们浸润 CNS 并产生 促炎细胞因子 IFNγ 和 IL-17 分别干扰初始 CD4+ T 细胞的发育。 通过分别基因删除转录因子 T-bet 和 RORγt 进入 Th1 和 Th17 细胞，保护 我们实验室的小鼠实验表明，CD4+ T 细胞需要钙离子的流入才能激活并发挥作用。 产生 IFNγ 和 IL-17 的能力由 CRAC（钙释放激活钙）介导。 位于细胞质膜上并由 ORAI1 蛋白形成的通道被激活。 由两种细胞内蛋白 STIM1 和 STIM2 以及这三种蛋白中任一种的缺失或突变 使用 ORAI1、STIM1 和基因缺失的小鼠来减弱 CRAC 通道功能和钙流入。 STIM2以及特定的CRAC通道抑制剂，我们发现Th1和Th17细胞更依赖 钙流入比其他 T 细胞具有引起自身免疫炎症的功能和能力。 具有特定 CRAC 通道抑制剂的小鼠或人类细胞减少了 IL-17 和 IFNγ 的产生，但 不以剂量依赖性方式影响 T 调节 (Treg) 细胞。 T细胞中的STIM1或STIM2，我们可以预防或改善EAE和CNS炎症的发展 重要的是，在 EAE 症状出现后，T 细胞中 ORAI1 或 STIM1 的缺失就停止了。 类似地，对患有 CRAC 通道的 EAE 小鼠进行治疗。 抑制剂还显着降低了疾病的严重程度，而没有明显的副作用。 钙内流是 EAE 中枢神经系统炎症的重要介质，其抑制可能是一种新的选择 用于治疗多发性硬化症的药物已被开发出来，但尚未经过测试。 它们在治疗 MS 和其他 T 细胞介导的自身免疫性疾病中的功效和安全性。 该应用具有双重意义：(1) 了解钙通过 CRAC 流入的分子机制 通道控制 CD4 T 细胞发育为致病性 T 细胞，特别是 Th1 和 Th17 细胞， 介导 CNS 炎症和 EAE/MS (2) 评估 CRAC 通道作为免疫治疗的药物靶点。 通过在复发缓解型 EAE 小鼠模型中研究它们在人类 T 细胞中的作用，研究 EAE 和最终 MS 以及对作为抗炎治疗潜在并发症的感染的免疫力。

项目成果

ORAI1, STIM1/2, and RYR1 shape subsecond Ca2+ microdomains upon T cell activation.

ORAI1、STIM1/2 和 RYR1 在 T 细胞激活后形成亚秒级 Ca2 微结构域。

• 发表时间: 2018
• 影响因子: 7.3
• 作者: Diercks, Björn;Werner, René;Weidemüller, Paula;Czarniak, Frederik;Hernandez, Lola;Lehmann, Cari;Rosche, Annette;Krüger, Aileen;Kaufmann, Ulrike;Vaeth, Martin;Failla, Antonio V;Zobiak, Bernd;Kandil, Farid I;Schetelig, Daniel;Ruthenbec
• 通讯作者: Ruthenbec