Regulation of encephalitogenic T cells by CRAC channels

CRAC 通道对致脑炎 T 细胞的调节

基本信息

批准号：
9981624
负责人：
STEFAN FESKE
金额：
$ 48.27万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
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/9981624
关键词：
Adverse effects Affect Animal Model Anti-Inflammatory Agents Attenuated Autoimmune Diseases Autoimmune Process 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 Estrogen receptor positive 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 autoreactive T cell autoreactivity brain cell chronic autoimmune disease cytokine in vivo inhibitor/antagonist interleukin-23 mouse model multiple sclerosis patient multiple sclerosis treatment novel strategies prevent transcription factor treatment strategy

项目摘要

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.

项目摘要多发性硬化症（MS）是一种自身免疫性疾病，其特征是中枢神经炎症系统（CNS）和大脑和脊髓的局部破坏，导致神经系统衰弱症状。 MS的疾病过程已在一种称为的动物模型中进行了广泛研究实验性自身免疫性脑脊髓炎（EAE），这极大地有助于开发几个批准的 MS的疗法。在MS和EAE中，CD4+辅助T细胞错误地识别由脑细胞为异物，并针对它安装自身免疫反应。这种炎症反应介导主要是由两个辅助T细胞的子集，即所谓的Th1和Th17细胞，它们浸润CNS并产生促炎细胞因子IFNγ和IL-17。干扰幼稚的CD4+ T细胞的发展通过转录因子T-BET和RORγT的遗传缺失，进入Th1和Th17细胞，可保护来自Eae的老鼠。我们的实验室表明，CD4+ T细胞需要钙离子的激活影响，并且产生IFNγ和IL-17的能力。钙影响由CRAC（钙释放激活钙）介导位于细胞质膜并由Orai1蛋白形成的通道。 Orai1被激活通过两种细胞内蛋白，即STIM1和Stim2，以及这三种蛋白质中的任何一个的缺失或诱变减弱CRAC通道功能和钙影响。将小鼠与Orai1，stim1和 STIM2以及特定的CRAC通道抑制剂，我们发现TH1和TH17细胞更依赖钙的影响比其他T细胞的功能和引起自身免疫注射的能力。 t的处理具有特定CRAC通道抑制剂的小鼠或人的细胞降低了IL-17和IFNγ的产生不影响T调节（Treg）细胞。以剂量依赖的方式。通过大致减弱两个ORAI1，在T细胞中的stim1或stim2，我们可以预防或改善EAE和CNS注入的发展老鼠。重要的是，EAE症状已经停止了T细胞中ORAI1或Stim1的删除停止疾病的进展。同样，对EAE使用CRAC通道开发的小鼠的治疗抑制剂还显着降低了疾病的严重程度，而没有明显的不良影响。这些数据表明钙影响是EAE中CNS注入的重要介质，其抑制作用可能是一种新选择用于治疗MS。已经开发了抑制CRAC通道的药物，但尚未测试它们在治疗MS和其他T细胞介导的自身免疫性疾病方面的效率和安全性。目标该应用是双重的：（1）了解钙通过CRAC影响的分子机制通道控制CD4 T细胞的发育到致病性T细胞中，尤其是TH1和TH17细胞，即介导CNS注入和EAE/MS。（2）评估CRAC通道作为免疫疗法的药物靶标通过研究复发EAE的小鼠模型，通过研究其在人类T细胞中的作用，最终是MS 在免疫学上，感染是抗炎疗法的潜在并发症。