STIM-Dependent Signaling in Cardiac Pathophysiology

心脏病理生理学中的 STIM 依赖性信号传导

• 批准号: 8509258
• 负责人: Salvatore Mancarella
• 金额: $ 10.51万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 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/8509258
• 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; 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; Property; 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; 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; response; sensor; skills

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赠款的目的是帮助Mancarella博士过渡到研究大学的稳定独立研究职位，在那里他可以在心血管领域进行研究。博士。唐纳德·吉尔（Donald Gill）和史蒂文·霍斯纳（Steven Houser）是两位著名的细胞信号和心血管科学专家，将在此过渡期间为曼卡雷拉（Mancarella）博士提供帮助。在他的博士后训练期间，候选人研究了CA2+传感器STIM1的生物物理特性。这些结果表明，刺激形成了专门的蛋白质网络，能够从近膜室捕获Ca2+信号，并以信号转换，以激活与信号源远端发生的基因转录。候选人认为，钙浓度微区域所需的stim1和stim2-与NFAT系统通信以调节心肌的病理生长。候选人将结合来自成像分析，离子通道，生物信息学，蛋白质分析和动物模型的数据，以建立蛋白质网络，以描述蛋白质之间的相互关系如何导致心脏肥大和心脏衰竭。在K99阶段，候选人将表征新生儿心肌细胞（AIM I）中刺激依赖性的“ Ca2+特征”，并隔离刺激激活的离子电流。为此，Mancarella博士将实施一系列技术，例如遗传学，活细胞成像和电生理学，以检查Stim1和Stim2在心脏中的功能作用。随后，（AIM 2）他将识别并表征刺激微生物群的蛋白质组成“相互作用”，该蛋白质控制着心脏的病理生长。蛋白质组学分析和生物信息学的整合将允许对近膜刺激刺激小区组成和心脏功能进行系统评估。为了实现他的目标，Mancarella博士将在Temple University蛋白质组学中心接受培训。在R00阶段，Mancarella博士将检验以下假设：刺激1和Stim2是心脏病理生长所必需的。实现此目标的实验方法是基于获得心脏特异性刺激的诱导双KO小鼠模型。这将通过CRE/LOX技术实现。所得的应变将允许在心脏中刺激刺激蛋白的时间和空间控制消融。该模型将作为验证AIM1和AIM2中的发现的体内模型，并向以下方向扩展：（a）检查刺激在成人心脏功能中的作用（b），以研究刺激在胸部主动力收缩（TAC）引起的压力超负荷中的作用，并检查，检查，心脏重塑和心脏重塑和心脏功能。评估将扩展到孤立的肌细胞，包括在局部和全球水平上使用共聚焦显微镜的外观，功能和CA2+动态。提出的多学科方法（生化和生理）将确保对候选人进行高质量的培训。