Compartmentation of Myocyte Adenosine Receptor Signaling

肌细胞腺苷受体信号传导的区室

• 批准号: 7670420
• 负责人: Robert D. Lasley
• 金额: $ 26.34万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-25 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7670420
• 关键词: Adenosine; Adenosine A1 Receptor; Agonist; Calcium; Cardiac Myocytes; Cardiac Surgery procedures; Cardiovascular Pathology; Cyclic GMP-Dependent Protein Kinases; Development; Diabetes Mellitus; Exhibits; Extracellular Signal Regulated Kinases; Funding; G-Protein-Coupled Receptors; Grant; Heart; Heart failure; Homeostasis; Hypertrophy; Ischemia; Knockout Mice; Laboratories; MAPK14 gene; Mediating; Mitochondria; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; Mus; Muscle Cells; Myocardial; Myocardial Ischemia; Myocardium; Nuclear; Opioid; Opioid Receptor; Pathway interactions; Pattern; Protein Isoforms; Protein Kinase; Proteins; Proto-Oncogene Proteins c-akt; Publishing; Purinergic P1 Receptors; Receptor Signaling; Reperfusion Injury; Reperfusion Therapy; Role; Signal Transduction; Subcellular Fractions; Therapeutic Agents; Ventricular; Ventricular Function; Ventricular Remodeling; Wild Type Mouse; in vivo; mitochondrial membrane; myocardial infarct sizing; preconditioning; receptor

DESCRIPTION (provided by applicant): A significant number of therapeutic agents for the treatment of ischemic heart disease target G protein coupled receptors (GPCR) and protein kinases. Up to four subtypes of one GPCR, the adenosine receptor, may be expressed in mammalian myocardium and all four subtypes have been shown to modulate protein kinase signaling. Published and preliminary findings from the applicant's laboratory indicate that adenosine A2a receptors modulate the cardioprotective effects of adenosine A1 receptor stimulation. Additional preliminary findings indicate that interactions between the adenosine A1 receptor and the 4-opioid receptor modulate the cardioprotective effects of each receptor. The cardioprotective effects of both adenosine A1 and 4-opioid receptors are dependent on multiple protein kinases. The overall hypothesis of this proposal is that the interactions between adenosine receptor subtypes and between adenosine and opioid receptors in mediating their cardioprotective effects are due to the modulation of subcellular protein kinase signaling. Specific Aim 1 will determine the role of interactions among adenosine receptor subtypes and opioid receptors in reduction of myocardial ischemia-reperfusion injury in intact hearts and isolated myocytes. Specific Aim 2 will determine whether the cardioprotective effects of adenosine receptor subtype and opioid receptor interactions are due to modulation of subcellular protein kinase signaling in intact myocardium and isolated cardiomyocytes. Specific Aim 3 will delineate the effects of interactions among adenosine receptor subtypes and opioid receptors on the modulation of myocyte contractility, intracellular calcium handling and mitochondrial function in isolated cardiomyocytes. Studies will be conducted in wild-type mice and mice with deletions of adenosine A1 receptors, A2a receptors, 4-opioid, and :-opioid receptors. Receptor signaling via the PKC, MAPK and AKT pathways will be examined in nuclear, cytosolic, mitochondrial and membrane subcellular fractions generated from normal and ischemic-reperfused isolated mouse hearts and isolated ventricular myocytes. Receptor- induced modulation of subcellular protein kinase signaling in intact hearts will be correlated with myocardial infarct size and ventricular function. Subcellular signaling in isolated myocytes will be correlated with myocyte contractility, intracellular calcium homeostasis and mitochondrial function. Although protection of ischemic myocardium appears to be mediated via activation of multiple protein kinases, conditions such as myocardial hypertrophy, heart failure, and ventricular remodeling following open heart surgery are also associated with increased protein kinase activity. The results of these in-depth studies of GPCRs and their interactions on subcellular protein kinase signaling in normal and ischemic myocardium may facilitate the development of new therapies for the treatment of the ischemic heart.

描述（由申请人提供）：大量用于治疗缺血性心脏病的治疗剂靶向G蛋白偶联受体（GPCR）和蛋白激酶。一种GPCR（腺苷受体）的多达四种亚型可在哺乳动物心肌中表达，并且所有四种亚型已显示出调节蛋白激酶信号传导。来自申请人实验室的公开和初步发现表明腺苷A2 a受体调节腺苷A1受体刺激的心脏保护作用。其他初步研究结果表明，腺苷A1受体和4-阿片受体之间的相互作用调节每个受体的心脏保护作用。腺苷A1和4-阿片受体的心脏保护作用依赖于多种蛋白激酶。该提议的总体假设是，腺苷受体亚型之间以及腺苷和阿片受体之间在介导其心脏保护作用中的相互作用是由于亚细胞蛋白激酶信号传导的调节。具体目标1将确定腺苷受体亚型和阿片受体之间的相互作用在减少完整心脏和离体心肌细胞的心肌缺血-再灌注损伤中的作用。具体目标2将确定腺苷受体亚型和阿片受体相互作用的心脏保护作用是否是由于完整心肌和分离心肌细胞中亚细胞蛋白激酶信号传导的调节。具体目标3将描述腺苷受体亚型和阿片受体之间的相互作用对离体心肌细胞中肌细胞收缩性、细胞内钙处理和线粒体功能的调节的影响。将在野生型小鼠和腺苷A1受体、A2 a受体、4-阿片类和：-阿片类受体缺失的小鼠中进行研究。通过PKC、MAPK和AKT途径的受体信号传导将在从正常和缺血再灌注的离体小鼠心脏和离体心室肌细胞产生的核、胞质、线粒体和膜亚细胞组分中进行检查。在完整心脏中受体诱导的亚细胞蛋白激酶信号转导的调节将与心肌梗死面积和心室功能相关。分离的肌细胞中的亚细胞信号传导将与肌细胞收缩性、细胞内钙稳态和线粒体功能相关。虽然缺血心肌的保护似乎是通过激活多种蛋白激酶介导的，但心脏直视手术后的心肌肥大、心力衰竭和心室重塑等疾病也与蛋白激酶活性增加有关。这些深入研究GPCR及其对正常和缺血心肌中亚细胞蛋白激酶信号传导的相互作用的结果可能有助于开发治疗缺血性心脏的新疗法。