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STIM-Dependent Signaling in Cardiac Pathophysiology

心脏病理生理学中的 STIM 依赖性信号传导

基本信息

批准号：
8712547
负责人：
Salvatore Mancarella
金额：
$ 10.29万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-02 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8712547
关键词：
Ablation Adult Affinity Animal Model Appearance Architecture Award Binding Biochemical Bioinformatics Calcineurin Calcium Calcium Channel Calmodulin Cardiac Cardiac Myocytes Cardiovascular system Cause of Death Cell membrane Cell physiology Chemicals Chest Confocal Microscopy Cre-LoxP Data Development Distal Electrocardiogram Electrophysiology (science)Ensure Environment Evaluation Experimental Designs Functional disorder Future Genes Genetic Genetic Transcription Genotype Gills Goals Grant Growth Health Heart Heart Diseases Heart Hypertrophy Heart failure Image Analysis Investigation Ion Channel Ions Knowledge Lead Life Luciferases Measures Mediating Membrane Mentors Modeling Movement Mus Muscle Cells Myocardium Neonatal Pathway Analysis Pathway interactions Phase Physiological Physiological Processes Play Positioning Attribute Protein Analysis Protein Kinase Proteins Proteome Proteomics Publishing Qualifying Reporter Research Research Personnel Role Route STIM1 gene Science Signal Transduction Source System Tamoxifen Techniques Technology Testing Therapeutic Agents Tissues Training Transducers United States Universities Validation Western World Work aorta constriction base biophysical properties calcium indicator career cellular imaging constriction in vivo in vivo Model inhibitor/antagonist interdisciplinary approach mouse model novel therapeutics patch clamp post-doctoral training pressure public health relevance response sensor skills

项目摘要

DESCRIPTION (provided by applicant): The purpose of this K99/R00 grant is to assist Dr. Mancarella in transitioning to a stable independent research position in a research University where he can conduct his research in the cardiovascular field. Drs. Donald Gill and Steven Houser, two renowned experts in cell signaling and cardiovascular science, will assist Dr. Mancarella during this transition. During his postdoctoral training the candidate has investigated the biophysical properties of the Ca2+ sensors STIM1. These results suggest that STIM forms specialized proteins network able to capture Ca2+ signals from the near-membrane compartment and convert it in signals that activate gene transcription which take place distal from the signal source. The candidate theorized that STIM1 and STIM2-are required for calcium-concentration microdomains which communicate with the NFAT system to regulate the pathological growth of the myocardium. The candidate will combine data from imaging analysis, ion channels, bioinformatics, protein analysis and animal models to build a protein network to describe how inter-relations between proteins lead to cardiac hypertrophy and heart failure. During the K99 phase, the candidate will characterize the STIM-dependent "Ca2+ signature" in neonatal cardiomyocytes (Aim I) and isolate the ionic currents activated by STIM. To this end Dr. Mancarella will implement a combination of techniques such as, genetics, live-cell imaging, and electrophysiology to examine the functional role of STIM1 and STIM2 in the heart. Subsequently, (Aim 2) he will identify and characterize the protein compositions of the STIM-microdomains "Interactome" that govern pathological growth of the heart. Proteomic analysis and bioinformatics integration will allow a systematic assessment of the near-membrane STIM microdomains composition and function in the heart. To achieve his Goals Dr. Mancarella will be trained at Temple University Proteomic Center. During the R00 phase, Dr. Mancarella will test the hypothesis that STIM 1 and STIM2 are required for pathological growth of the heart. The experimental approach to this aim is based on obtain a cardiac specific STIM1/2 inducible double KO mouse model; this will be achieved by Cre/Lox technology. The resultant strain will allow a temporally and spatially controlled ablation of the STIM proteins in the heart. This model will serve as an in vivo model to validate the findings in aim1 and aim2 and expand towards: (a) Examine the role of STIM in adult cardiac functions (b) To investigate the role of STIM during pressure overload induced by thoracic aortic constriction (TAC) and examine, cardiac remodeling and cardiac functions. The evaluation will be extended to the isolated myocytes, including appearance, functions and Ca2+ dynamic at local as well as global level with the use of confocal microscopy. The multidisciplinary approach (biochemical and physiological) proposed will ensure a high quality training of the candidate.

描述（由申请人提供）：这项K99/R00资助的目的是帮助Mancarella博士在研究型大学过渡到一个稳定的独立研究职位，在那里他可以进行心血管领域的研究。Drs。Donald Gill和Steven Houser是两位著名的细胞信号传导和心血管科学专家，他们将在这一过渡期间协助Mancarella博士。在他的博士后培训期间，候选人研究了Ca2+传感器STIM1的生物物理特性。这些结果表明，STIM形成了特殊的蛋白质网络，能够捕获来自近膜室的Ca2+信号，并将其转化为激活信号源远端的基因转录的信号。候选人的理论认为，STIM1和stim2是钙浓度微域所必需的，这些微域与NFAT系统通信，以调节心肌的病理生长。该候选人将结合成像分析、离子通道、生物信息学、蛋白质分析和动物模型的数据，建立一个蛋白质网络，描述蛋白质之间的相互关系如何导致心脏肥厚和心力衰竭。在K99期，候选人将在新生儿心肌细胞（Aim I）中表征STIM依赖的“Ca2+特征”，并分离由STIM激活的离子电流。为此，Mancarella博士将结合遗传学、活细胞成像和电生理学等技术来检查STIM1和STIM2在心脏中的功能作用。随后，（目标2）他将识别和表征控制心脏病理生长的stim微域“相互作用组”的蛋白质组成。蛋白质组学分析和生物信息学集成将允许对心脏近膜STIM微结构域的组成和功能进行系统评估。为了实现他的目标，Mancarella博士将在天普大学蛋白质组学中心接受培训。在R00阶段，Mancarella博士将测试stim1和STIM2是心脏病理生长所必需的假设。实现这一目标的实验方法是建立心脏特异性STIM1/2诱导的双KO小鼠模型；这将通过Cre/Lox技术实现。由此产生的菌株将允许在时间和空间上控制心脏中STIM蛋白的消融。该模型将作为体内模型来验证aim1和aim2的研究结果，并向以下方向扩展：(a)研究STIM在成人心功能中的作用(b)研究STIM在胸主动脉收缩（TAC）引起的压力过载中的作用，并检查心脏重塑和心功能。评估将扩展到分离的肌细胞，包括外观，功能和Ca2+动态在局部和全局水平使用共聚焦显微镜。提出的多学科方法（生化和生理）将确保候选人的高质量培训。