Molecular Mechanisms of lon Channels in T Lymphocytes

T淋巴细胞lon通道的分子机制

基本信息

批准号：
10408818
负责人：
MICHAEL D CAHALAN
金额：
$ 55万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
1978
资助国家：
美国
起止时间：
1978-09-15 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10408818
关键词：
Acceleration Actins Adoptive Transfer Alzheimer&apos s Disease Antigen Presentation Antigens Astrocytes Autoimmune Autoimmune Diseases Behavior Binding Blood Platelets CD8-Positive T-Lymphocytes Calcium Calcium Channel Calcium Signaling Calcium ion Calmodulin Cell Proliferation Cell membrane Cell physiology Cells Characteristics Chimeric Proteins Chronic Coin Complement Cytoplasmic Granules Cytoskeleton Data Dentition Development Disease Drug or chemical Tissue Distribution Electrophysiology (science)Endothelial Cells Environment Event Family Feedback Fluorescence Fluorescence Microscopy Gene Expression Goals Graft Rejection Heterogeneity Human Image Immune response Immune system Immunologic Surveillance In Situ Individual Infection Inflammatory Insulin-Dependent Diabetes Mellitus Ion Channel Ions Knock-in Knock-in Mouse Knockout Mice Lead Lymphocyte Mediating Membrane Membrane Potentials Microglia Microscopy Microtubules Molecular Monitor Multiple Sclerosis Mus Names Natural Killer Cells Neurons Optics Output Patch-Clamp Techniques Patients Pattern Pharmacology Phosphorylation Physiological Processes Piezo 1 ion channel Play Property Protein Isoforms Proteins RNA interference screen Reporting Role STIM1 gene Salivary Glands Severe Combined Immunodeficiency Signal Transduction Site Skeletal Muscle Sweat Glands System T-Cell Receptor T-Lymphocyte Tail Testing Tissues Transgenic Mice Variant Vascular Smooth Muscle Visualization base cell motility cell type confocal imaging cytokine disorder prevention drug development fluorescence imaging in vivo insight lymph nodes mast cell novel optical imaging overexpression patch clamp ratiometric response single molecule targeted treatment therapeutic target tool two-photon

项目摘要

Project Summary / Abstract Store-operated Ca2+ entry (SOCE) underlies numerous cellular processes throughout the body and initiates signaling cascades in T lymphocytes that cause changes in motility, secretion of cytolytic granules, cytokine release, and cell proliferation. The channels that underlie SOCE have been identified recently through RNA interference (RNAi) screening as a conserved family of four transmembrane-spanning proteins named Orai that are activated by STIM proteins in the ER membrane. Isoforms of these proteins are expressed throughout the body in a tissue-specific manner. Important cellular functions of Orai1 have been identified in lymphocytes, microglia, mast cells, blood platelets, sweat and salivary glands, dentition, vascular smooth muscle, endothelial cells, skeletal muscle, microglia, astrocytes, and neurons. In the immune system, STIM1 and Orai1 mediate antigen-induced Ca2+ signaling, motility inhibition at the site of antigen presentation, secretion of cytolytic granules by CD8+ T cells and NK cells, and gene expression responses that lead to cytokine release and cell proliferation. STIM and Orai proteins are being developed as targets for treatment of autoimmune diseases and prevention of transplant rejection. Our overall goal is to understand how Orai channels function at the molecular and cellular level. Orai channels in the plasma membrane are unrelated to other known ion channels and have unusual characteristics that distinguish them, including a very high degree of selectivity for Ca2+, low single-channel conductance, and activation by binding of a small cytosolic domain of the STIM protein. Moreover, the human Orai1 and Orai3 proteins differ in their activation requirements and tissue distribution. In this project, we have three goals. We seek to understand at molecular, subcellular, and cellular levels: 1) how Orai1 is activated by STIM1 at the single-channel level; 2) how Orai1 signals locally in puncta to generate localized Ca2+ signals; 3) how local Ca2+ signals modulate T cell motility, turning behavior and stopping during immune surveillance. To accomplish these Aims, we have developed and continue to develop new tools for monitoring local Ca2+ signals that will be broadly applicable. Our studies will include electrophysiological analysis, optical imaging of Ca2+ flux through Orai1 channels, and two-photon imaging of in situ cellular motility. Overall, our project will provide fundamental insights into the Orai1 proteins that are currently being targeted for treatment of autoimmune disorders, chronic inflammatory conditions, and neurodegerative diseases such as Alzheimer’s.

项目摘要 /摘要商店经营的Ca2+进入（SOCE）是整个身体的许多细胞过程，并发起 T淋巴细胞中的信号级联反应会导致运动变化，细胞溶剂的分泌，细胞因子的分泌释放和细胞增殖。最近已经通过RNA确定了SOCE基础的渠道干涉（RNAi）筛查是一个保守的四个跨膜跨膜蛋白的家族，称为Orai 被ER膜中的刺激蛋白激活。这些蛋白质的同工型在整个体内表达以组织特异性的方式。 ORAI1的重要细胞功能已在淋巴细胞，小胶质细胞中鉴定肥大细胞，血小板，汗水和唾液手套，牙齿，血管平滑肌，内皮细胞，骨骼肌肉，小胶质细胞，星形胶质细胞和神经元。在免疫系统中，STIM1和ORAI1中位抗原诱导 Ca2+信号传导，抗原表现部位的运动性抑制，CD8+ T细胞对细胞溶液的分泌 NK细胞以及导致细胞因子释放和细胞增殖的基因表达反应。 Stim和Orai 正在开发蛋白质作为治疗自身免疫性疾病和预防移植的靶标拒绝。我们的总体目标是了解Orai通道在分子和细胞水平上的功能。质膜中的Orai通道与其他已知离子通道无关，并且具有不寻常的区分它们的特征，包括CA2+的高度选择性，低单通道电导和通过结合刺激蛋白的小胞质结构域而激活。而且，人类Orai1 ORAI3蛋白的激活需求和组织分布有所不同。在这个项目中，我们有三个目标。我们寻求在分子，亚细胞和细胞水平上理解：1）在单通道级； 2）如何在puncta中本地信号产生局部Ca2+信号； 3）局部CA2+如何信号调节T细胞运动，在免疫监视过程中转动行为和停止。完成这些目的，我们已经开发并继续开发新工具来监视本地CA2+信号适用的。我们的研究将包括电生理分析，通过Orai1的Ca2+通量的光学成像原位细胞运动的通道和两光子成像。总体而言，我们的项目将提供基本的见解进入目前用于治疗自身免疫性疾病的ORAI1蛋白炎症状况和神经变性疾病，例如阿尔茨海默氏症。

项目成果

期刊论文数量（33）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Calcium Imaging in T Lymphocytes: a Protocol for Use with Genetically Encoded or Chemical Ca2+ Indicators.

T 淋巴细胞钙成像：与基因编码或化学 Ca2 指示剂一起使用的方案。

DOI：
10.21769/bioprotoc.4170
发表时间：
2021
期刊：
Bio-protocol
影响因子：
0.8
作者：
Jairaman,Amit;Cahalan,MichaelD
通讯作者：
Cahalan,MichaelD

Chemical modification of sodium channel surface charges in frog skeletal muscle by trinitrobenzene sulphonic acid.

三硝基苯磺酸对青蛙骨骼肌钠通道表面电荷的化学修饰。

DOI：
10.1113/jphysiol.1981.sp013975
发表时间：
1981
期刊：
The Journal of physiology
影响因子：
0
作者：
Cahalan,MD;Pappone,PA
通讯作者：
Pappone,PA

STIM1, an essential and conserved component of store-operated Ca2+ channel function.

DOI：
10.1083/jcb.200502019
发表时间：
2005-05-09
期刊：
JOURNAL OF CELL BIOLOGY
影响因子：
7.8
作者：
Roos, Jack;DiGregorio, Paul J;Yeromin, Andriy V;Ohlsen, Kari;Lioudyno, Maria;Zhang, Shenyuan;Safrina, Olga;Kozak, J Ashot;Wagner, Steven L;Cahalan, Michael D;Velicelebi, Gonul;Stauderman, Kenneth A
通讯作者：
Stauderman, Kenneth A

Orai3 TM3 point mutation G158C alters kinetics of 2-APB-induced gating by disulfide bridge formation with TM2 C101.