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Calcium Signaling Roles of STIM1 and STIM2 in Smooth Muscle

STIM1 和 STIM2 在平滑肌中的钙信号传导作用

基本信息

批准号：
9236203
负责人：
Donald L Gill
金额：
$ 40.72万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-05 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9236203
关键词：
Address Aging Angioplasty Animal Model Animals Asthma Atherosclerosis Blood Vessels Calcium Signaling Cardiovascular Diseases Cell Proliferation Cell physiology Coupling Defect Development Effectiveness Electrophysiology (science)Endoplasmic Reticulum Event Fluorescence Resonance Energy Transfer Gene Deletion Genes Goals Growth Hypertension Image In Vitro Knock-out Knowledge Lead Lung Mediating Mitogens Modeling Modification Molecular Molecular Probes Mus Muscle Cells Mutation Natural regeneration Pattern Perinatal Phenotype Process Property Proteins Resolution Role STIM1 gene Signal Transduction Smooth Muscle Smooth Muscle Myocytes Structure System Transcriptional Regulation Transducers Vascular Diseases Vascular Smooth Muscle Work airway remodeling angiogenesis cell growth cell injury cell motility genetic approach in vivo injury and repair innovation insight knockout animal operation predictive modeling prevent protein B public health relevance response response to injury restenosis sensor transcription factor tumor tumor growth voltage

项目摘要

DESCRIPTION (provided by applicant): Calcium signals are vital in controlling cellular events. In smooth muscle cells (SMC), short term Ca2+ signals control contractile responses whereas longer term Ca2+ signals regulate cell growth and proliferation through Ca2+-mediated transcriptional control. The proposed studies focus on the two centrally important Ca2+ signal transducers, STIM1 and STIM2. STIM proteins are finely-tuned endoplasmic reticulum (ER) Ca2+ sensors. STIM proteins are triggered to self-associate and translocate into specialized ER-PM junctions within whom the STIM proteins gate the highly Ca2+ selective "store-operated" Orai channels. STIM proteins also target "voltage-operated" L-type Ca2+ channels (LTCC) and exert reciprocal control over these two channel targets, activating Orai but deactivating LTCC, likely important to SMC growth phenotype change in which LTCC is lost and STIM-Orai signaling predominates. While STIM1 has been intensively examined, a major challenge is to understand the role of the poorly-studied STIM2 protein: how differences in its structure, function and expression lead to powerful signaling and phenotypic changes. While whole animal knockout of either STIM1 or STIM2 are lethal, we generated conditional SMC-targeted deletions of STIM1, STIM2, and STIM+STIM2 in mice. The SMC-specific STIM1-KO results in poorly-developed animals dying early, with major defects in the ability of SMC to undergo growth transition, in vivo and in vitro. The SMC conditional STIM1/STIM2 double knockout is perinatally lethal indicating important yet distinct roles of STIM1 and STIM2 in SMC function and growth transition. Using these systems, we have two independent but inter-dependent specific aims 1. To understand the mechanisms and distinctions between STIM1 and STIM2 activation, and to define their molecular coupling to target Ca2+ channels. Using mutational modifications, high resolution Ca2+ and FRET imaging, and electrophysiology, we aim to define the molecular basis of sensing and target channel coupling for STIM1, defining mechanistically important distinctions in the poorly studied STIM2 protein. Building on new structural insights and major functional distinctions between STIM1 and STIM2, our studies focus on defining the interactions through which the two STIM proteins interact with and gate Orai and LTCC target channels. 2. To understand the distinct roles of STIM1 and STIM2 in mediating mitogen-induced growth transition of SMC from the quiescent to proliferative phenotype. Using our SMC-targeted STIM1 and STIM2 null-back- ground mice together with molecular probes to specifically modify STIM1- and STIM2-mediated target channel coupling, we aim to define the differential effectiveness of STIM1 and STIM2 in mediating SMC growth transition. Studies will determine how changes in STIM1 and STIM2 expression and distinctions in their activation and channel coupling, lead to distinct profiles of Ca2+ signals, NFAT activation, and altered growth transition. Our studies apply: (a) new knowledge on the structure and channel-coupling of STIM proteins, (b) innovative STIM1 and STIM2 gene-deletion animal models, (c) innovative probes to assess STIM1 and STIM2 target channel coupling - to provide new understanding of the crucial role of STIM1 in controlling SMC growth transition. Our goals also address a fundamental paucity of information on understanding the mechanism, action and role of STIM2. Information from these studies is crucial to understanding how vascular SMC injury responses occur and how they may be controlled. Our model predicts that distinct patterns of STIM1 and STIM2 expression can be a determining factor in whether SMC undergo phenotype change. Hence the work has fundamental importance in understanding and preventing the SMC growth changes that underlie major vascular diseases including atherosclerosis, hypertension, and arterial restenosis, and SMC phenotype change that underlies lung responses in asthma and angiogenesis that supports growth of tumors.

描述(由申请人提供)：钙信号在控制细胞事件中至关重要。在平滑肌细胞(SMC)中，短期的Ca~(2+)信号控制收缩反应，而长期的Ca~(2+)信号通过Ca~(2+)介导的转录调控调节细胞的生长和增殖。建议的研究集中在两个中央重要的钙信号转导系统，STIM1和STIM2。STIM蛋白是微调的内质网(ER)钙离子感受器。STIM蛋白被触发自结合并转移到专门的ER-PM连接中，在这个连接中，STIM蛋白控制着高度选择性的钙离子“储存操作”的Orai通道。Stim蛋白还以“电压操纵型”的L钙通道(LTCC)为靶点，并对这两个通道靶点实施相互控制，激活Orai但抑制LTCC，这可能对LTCC缺失、STIM-Orai信号占优势的SMC生长表型改变很重要。虽然STIM1已经被深入研究，但一个主要的挑战是了解研究较少的STIM2蛋白的作用：它的结构、功能和表达的差异如何导致强大的信号和表型变化。虽然整个动物敲除STIM1或STIM2都是致命的，但我们在小鼠中产生了有条件的SMC靶向缺失STIM1、STIM2和STIM+STIM2。SMC特异性STIM1-KO导致发育不良的动物过早死亡，体内和体外SMC经历生长过渡的能力存在重大缺陷。SMC条件性STIM1/STIM2双基因敲除对围产儿是致命的，提示STIM1和STIM2在SMC功能和生长转变中具有重要而独特的作用。利用这些系统，我们有两个相互独立但又相互依赖的特异性目标：1.了解STIM1和STIM2激活的机制和区别，并确定它们与靶向钙通道的分子偶联。利用突变修饰、高分辨率钙离子和FRET成像以及电生理学，我们的目标是定义STIM1的传感和靶通道耦合的分子基础，定义研究较少的STIM2蛋白在机制上的重要区别。在STIM1和STIM2之间新的结构洞察和主要功能差异的基础上，我们的研究重点是定义这两个STIM蛋白与AND GATE ORAI和LTCC靶向通道的相互作用。2.了解STIM1和STIM2在丝裂原诱导的SMC从静止期向增殖期转化中的不同作用。使用我们的SMC靶向STIM1和STIM2零基础小鼠以及分子探针来特异性地修改STIM1和STIM2介导的靶通道偶联，我们的目标是确定STIM1和STIM2在介导SMC生长转变方面的不同有效性。研究将确定STIM1和STIM2表达的变化以及它们在激活和通道耦合方面的差异，导致不同的钙信号、NFAT激活和改变的生长转变。我们的研究应用于：(A)对STIM蛋白的结构和通道偶联的新认识，(B)创新的STIM1和STIM2基因缺失动物模型，(C)评估STIM1和STIM2靶通道偶联的创新探针--为STIM1在控制SMC生长过渡中的关键作用提供新的理解。我们的目标还解决了关于理解STIM2的机制、作用和作用的基本信息匮乏的问题。来自这些研究的信息对于了解血管SMC损伤反应如何发生以及如何控制它们至关重要。我们的模型预测，STIM1和STIM2表达的不同模式可能是决定SMC是否发生表型变化的一个决定因素。因此，这项工作对于了解和预防主要血管疾病(包括动脉粥样硬化、高血压和动脉再狭窄)背后的SMC生长变化，以及哮喘中肺反应和支持肿瘤生长的血管生成的SMC表型变化具有重要意义。

项目成果

期刊论文数量（5）

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 store refilling and STIM activation in STIM- and Orai-deficient cell lines.

STIM 和 Orai 缺陷细胞系中的钙储存补充和 STIM 激活。

DOI：
10.1007/s00424-018-2165-5
发表时间：
2018-10
期刊：
Pflugers Archiv : European journal of physiology
影响因子：
0
作者：
Zheng S;Zhou L;Ma G;Zhang T;Liu J;Li J;Nguyen NT;Zhang X;Li W;Nwokonko R;Zhou Y;Zhao F;Liu J;Huang Y;Gill DL;Wang Y
通讯作者：
Wang Y

Calcium signals tune the fidelity of transcriptional responses.