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 该项目的目的是确定钙通道亚基在T淋巴细胞中的作用（T 细胞）和免疫反应。 T细胞是白细胞通过杀死在病毒感染期间起关键作用的白细胞 感染病毒的细胞并促进病毒中和抗体的产生。 T细胞的功能 取决于形成细胞膜孔的钙通道并促进钙的流入 细胞。在细胞内部，钙充当与各种蛋白质结合并促进的信号分子 T细胞的激活，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编码的称为CRAC通道的钙通道的功能 刺激基因。最近的报道表明，其他钙通道也可能介导钙涌入和T细胞 功能。这些通道包括电压门控钙通道，在心脏中进行了良好的研究 他们在响应电刺激的情况下调节细胞功能的大脑。该提议中研究的基因是 CACNB1编码电压门控钙通道的四个已知调节亚基之一。 CACNB1 对于通道功能的不同方面至关重要，包括通道运输，激活和灭活。过去的 CACNB1的研究集中在可激发的细胞（例如神经元）上，但CaCNB1也在t中表达 其在免疫反应中作用的细胞仍然未知。最近，两种孔形成的遗传研究 电压门控钙通道和几个调节亚基的亚基显示了t功能的改变 缺乏这些通道亚基或表达突变体形式的细胞。特别是调节的突变 据报道，CaCNB2，B3和B4基因会影响胸腺中T细胞的发展和稳态的发育 脾。 CaCNB1在T细胞中的作用尚未研究 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，这部分是由于它们在T细胞中不明确的激活机制， 不是电气兴奋的。我们最近在活小鼠的功能基因组筛选中确定了CACNB1 作为调节小鼠病毒感染T细胞生长的几种离子通道之一。在这个 应用，我们建议进一步表征CaCNB1在T细胞功能和免疫反应中的作用。 具体而言，我们将研究CACNB1缺失如何影响钙信号传导，钙通道功能和 T细胞生长，产生免疫调节性细胞因子并杀死病毒感染细胞的能力。此外，我们 将通过感染T 细胞缺乏病毒和分析其免疫反应的CACNB1。拟议的研究将使新的 关于T细胞中CaCNB1和电压门控钙通道功能以及免疫反应的光。 CACNB1是T细胞中最高，特别表达的调节亚基，可能代表一个新的 治疗自身免疫性和炎症的药物靶标。