Calcium channel CACNB1 in T cell function and immunity

钙通道 CACNB1 在 T 细胞功能和免疫中的作用

• 批准号: 9811165
• 负责人: STEFAN FESKE
• 金额: $ 16.95万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2021-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9811165
• 关键词: Adoptive Transfer; Affect; Antibodies; Antibody Affinity; Antiviral Agents; Apoptosis; Autoimmune Diseases; Autoimmunity; B-Lymphocytes; Binding; Biological; Brain; CD4 Positive T Lymphocytes; Calcium; Calcium Channel; Calcium Signaling; Cardiac Myocytes; Cell membrane; Cell physiology; Cells; Clonal Expansion; Colitis; Complex; Coupling; Custom; Data; Disease; Disputes; Electric Stimulation; Genes; Genetic study; Goals; Growth; Guide RNA; Heart; Helper-Inducer T-Lymphocyte; Homeostasis; Image; Immune; Immune response; Immunity; Immunoglobulin G; Immunoglobulin M; In Vitro; Infection; Inflammation; Ion Channel; Knockout Mice; Leukocytes; Libraries; Light; Lymphocyte Function; Lymphocytic choriomeningitis virus; Measurement; Mediating; Membrane; Methods; Movement; Mus; Mutation; Neurons; Physiological; Play; Production; Proliferation Marker; Proteins; Reagent; Reporting; Research; Rheumatoid Arthritis; Role; Signal Transduction; Signaling Molecule; Spleen; Splenocyte; Structure of germinal center of lymph node; T cell response; T cell therapy; T-Cell Activation; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Testing; Thymus Gland; Transgenic Organisms; Validation; Viral; Virus; Virus Diseases; antiviral immunity; cell mediated immune response; cytokine; experimental study; functional genomics; in vivo; insight; knock-down; migration; mutant; neutralizing antibody; new therapeutic target; novel; response; retroviral transduction; small hairpin RNA; small molecule; targeted treatment; tool; trafficking; transmission process; tumor; voltage; voltage gated channel

Project Summary The goal of this project is to determine the role of a poorly studied calcium channel subunit in T lymphocytes (T cells) and immune responses. T cells are white blood cells that play a critical role during viral infection by killing virus-infected cells and promoting the production of virus-neutralizing antibodies. The function of T cells depends on calcium channels that form pores in the cell membrane and facilitate the influx of calcium into cells. Inside cells, calcium functions as a signaling molecule that binds to various proteins and promotes the activation of T cells, for instance by regulating the expression of immune modulatory genes. Our lab has extensively studied the function of calcium channels called CRAC channels that are encoded by ORAI and STIM genes. Recent reports suggest that other calcium channels may also mediate calcium influx and T cell function. Among those channels are voltage-gated calcium channels, which are well studied in the heart and brain where they regulate cell function in response to electrical stimulation. The gene studied in this proposal is CACNB1, which encodes one of four known regulatory subunits of voltage-gated calcium channels. CACNB1 is critical for different aspects of channel function including channel trafficking, activation and inactivation. Past research on CACNB1 has focused on excitable cells such as neurons, but CACNB1 is also expressed in T cells, where its role in immune responses remains unknown. Recently, genetic studies of two pore-forming subunits of voltage-gated calcium channels and several regulatory subunits have shown altered function of T cells that lack these channel subunits or express mutant forms. In particular, mutation of the regulatory CACNB2, B3 and B4 genes were reported to affect T cells development in the thymus and homeostasis in the spleen. The role of CACNB1 in T cells has not been investigated yet although it is the most highly expressed of the four regulatory subunits in T cells. Overall, the role of voltage-gated calcium channels in T cells and immune responses remains disputed, which in part is due to their unclear activation mechanism in T cells, which are not electrically excitable. We recently identified CACNB1 in a functional genomics screen in live mice as one of several ion channels that regulates the growth of T cells in response to viral infection of mice. In this application, we propose to further characterize the role of CACNB1 in T cell function and immune responses. Specifically, we will study how deletion of CACNB1 affects calcium signaling, calcium channel function and the ability of T cells to grow, produce immune modulatory cytokines and kill virus-infected cells. Furthermore, we will investigate the consequences of CACNB1 deficiency for immunity to infection by infecting mice whose T cells lack CACNB1 with viruses and analyzing their immune responses. The proposed research will shed new light on the function of CACNB1 and voltage-gated calcium channels in T cells and immune responses. CACNB1 is the most highly and specifically expressed regulatory subunit in T cells and may represent a new drug target for the treatment of autoimmunity and inflammation.

项目摘要 这个项目的目标是确定一个研究较少的钙通道亚单位在T淋巴细胞(T)中的作用 细胞)和免疫反应。T细胞是一种白细胞，它在病毒感染过程中通过杀死 并促进病毒中和抗体的产生。T细胞的功能 依赖于在细胞膜上形成毛孔并促进钙内流的钙通道 细胞。在细胞内，钙起信号分子的作用，与各种蛋白质结合，促进 激活T细胞，例如通过调节免疫调节基因的表达。我们的实验室有 广泛研究了由ORAI和ORAI编码的称为CRAC通道的钙通道的功能 Stim基因。最近的报道表明，其他钙通道也可能介导钙内流和T细胞 功能。这些通道中有电压门控钙通道，它在心脏和心脏中得到了很好的研究 它们调节细胞功能以响应电刺激的大脑。这项提案中研究的基因是 CACNB1编码已知的电压门控钙通道四个调节亚基之一。CACNB1 对于渠道功能的不同方面至关重要，包括渠道贩运、激活和失活。过去时 对CACNB1的研究主要集中在神经元等可兴奋细胞上，但CACNB1也在T细胞中表达 细胞，其在免疫反应中的作用尚不清楚。近年来，两种成孔作用的遗传学研究 电压门控性钙通道的亚基和几个调节亚基显示出T的功能改变 缺乏这些通道亚单位或表达突变形式的细胞。特别是，监管机构的突变 CACNB2、B3和B4基因被报道影响胸腺T细胞的发育和体内的稳态。 脾。CACNB1在T细胞中的作用尚未被研究，尽管它是T细胞中表达最高的 T细胞中的四个调节亚基。总体而言，电压门控钙通道在T细胞和 免疫反应仍然存在争议，部分原因是它们在T细胞中的激活机制不清楚， 它们是不能电刺激的。我们最近在活体小鼠的功能基因组学筛查中发现了CACNB1 作为调节T细胞生长以应对病毒感染的几种离子通道之一。在这 为了进一步研究CACNB1在T细胞功能和免疫反应中的作用，我们建议对CACNB1进行进一步的研究。 具体地说，我们将研究CACNB1缺失如何影响钙信号转导、钙通道功能和 T细胞生长、产生免疫调节细胞因子和杀死感染病毒的细胞的能力。此外，我们 将通过感染T细胞亚群的小鼠来研究CACNB1缺陷对感染免疫的影响 细胞缺乏CACNB1与病毒和分析其免疫反应。拟议中的研究将带来新的 CACNB1和电压门控钙通道在T细胞和免疫反应中的作用。 CACNB1是T细胞中表达最高和特异性最高的调节亚基，可能代表着一种新的 治疗自身免疫和炎症的药物靶点。