REGULATION OF MYOCARDIAL PHOSPHOLIPASES AND LIPASES IN DIABETIC MYOCARDIUM

糖尿病心肌中心肌磷脂酶和脂肪酶的调节

基本信息

批准号：
9065644
负责人：
RICHARD W GROSS
金额：
$ 75.54万
依托单位：
WASHINGTON UNIVERSITY
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：
2013-06-01 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9065644
关键词：
Ablation Active Sites Acute Acyl Coenzyme A Acyltransferase Arachidonic Acids Attenuated Bioenergetics Biological Biological Assay Calcium Calcium ion Cardiac Cardiac Myocytes Cause of Death Cell membrane Cessation of life Charge Chemicals Chronic Coenzyme A-Transferases Complex Coupling Diabetes Mellitus Disease Eicosanoids Employee Strikes Endocannabinoids Energy Intake Enzymes Fatty Acids Fatty acid glycerol esters Functional disorder Generations Genetic Health Heart Heart Diseases Heart failure Homeostasis Hydrolysis Infarction Injury Ischemia Knock-out Knockout Mice Lead Lipase Lipids Lysophosphatidylcholines Lysophospholipase Mass Spectrum Analysis Mediating Mediator of activation protein Membrane Membrane Structure and Function Mitochondria Molecular Monoglycerides Mus Mutate Myocardial Myocardial Contraction Myocardial Ischemia Myocardium Necrosis PLA2G6 gene Pathologic Pathway interactions Phospholipase Phosphorylation Phosphorylation Site Physiological Physiological Processes Production Protein Isoforms Proteins Regulation Reperfusion Therapy Role Serine Signal Transduction Signaling Molecule Site-Directed Mutagenesis Societies Stable Isotope Labeling Structure Sudden Death Testing Transacylase Transgenic Organisms Triglycerides Ventricular Arrhythmia calmodulin-dependent protein kinase II diabetic diabetic cardiomyopathy diabetic patient heart cell hemodynamics insulin signaling interdisciplinary approach loss of function lysophosphatidic acid metabolomics mitochondrial dysfunction mitochondrial permeability transition pore novel novel therapeutics transacylation

项目摘要

DESCRIPTION (provided by applicant): Diabetic cardiomyopathy is a complex disorder that emanates from the chronic and excessive use of fatty acids to fuel contractile function in diabetic myocardium due to the lack of insulin signaling. However, the nearly exclusive use of fatty acids for fuel in diabetic myocardium results in widespread metabolomic dysregulation that precipitates multiple deleterious alterations in membrane structure and function. Consequences of these membrane-mediated abnormalities in diabetic myocardium include hemodynamic compromise, defective excitation-contraction coupling and mitochondrial dysfunction that collectively conspire to promote the progression of heart failure in diabetic patients. Moreover, the profound alterations in substrate utilization in diabetic myocardium result in the accumulation of multiple dysregulated metabolites that lead to maladaptive alterations in interwoven cardiac myocyte signaling networks. Previously, through genetic, pharmacologic and chemical biological approaches, we have identified three major phospholipases and lipases in myocardium iPLA2ß (PNPLA9), iPLA2γ (PNPLA8), and iPLA2ζ (PNPLA2; ATGL) that likely serve as principal mediators of myocardial hemodynamic dysfunction, electrophysiologic alterations and maladaptive remodeling in diabetic myocardium. Recently, we demonstrated that iPLA2γ and its downstream signaling metabolites are key regulators of the mitochondrial permeability transition pore which is responsible for necrosis, necroptosis, and electrical instability in diabetic myocardium subjected to ischemia. Accordingly, in Specific Aim 1, we will use the novel cardiac myocyte specific iPLA2γ conditional knock out mouse we generated to determine if iPLA2γ loss of function attenuates acute ischemic injury, electrophysiologic instability and the maladaptive generation of lipid 2nd messengers in diabetic myocardium. Furthermore, we demonstrated that exposure of mitochondria to calcium ion results in the activation of iPLA2γ leading to the release of arachidonic acid, 2-arachidonoyl lysophosphatidylcholine, and the subsequent production of multiple downstream biologically active lipid 2nd messengers. Accordingly, iPLA2γ-dependent alterations in lipid 2nd messenger production will be examined employing integrative mass spectrometric platforms we developed in conjunction with the cardiac myocyte specific iPLA2γ loss of function mouse. In Specific Aim 2, we will determine the molecular mechanisms through which acyl-CoA facilitates CaMKII phosphorylation and activation of iPLA2ß. The activating phosphosite(s) will be identified, mutated and their mechanistic importance in CaMKII-mediated activation of iPLA2ß in diabetic myocardium and diabetic myocardium rendered ischemic will be explored. In Specific Aim 3, the role(s) of iPLA2ζ (ATGL;PNPLA2) in catalyzing the bidirectional flux of lipids through triglyceride hydrolysis, transacylation and acyltransferase activities will e determined. The participation of iPLA2ζ in generating lipid 2nd messengers in diabetic myocardium will be examined using cardiac myocyte specific iPLA2ζ null mice and the effects of iPLA2ζ genetic ablation on myocardial function in the diabetic state will be explored. Collectively, these studies are a synergistic multidisciplinary approach to identify the chemical mechanisms mediating diabetic cardiomyopathy.

描述（由申请人提供）：糖尿病心肌病是一种复杂的疾病，由于缺乏胰岛素信号而长期过量使用脂肪酸来促进糖尿病心肌的收缩功能。然而，几乎完全使用脂肪酸作为燃料。糖尿病心肌中的广泛代谢失调导致糖尿病心肌膜结构和功能的多种有害改变。心肌中的血流动力学受损、兴奋-收缩耦合缺陷和线粒体功能障碍共同促进了糖尿病患者心力衰竭的进展。此外，糖尿病心肌中底物利用的深刻改变导致了积累。此前，通过遗传、药理学和化学生物学方法，我们已经确定了心肌 iPLA2ß (PNPLA9)、iPLA2γ (PNPLA8) 和 iPLA2ze 中的三种主要磷脂酶和脂肪酶。 PNPLA2；ATGL）可能是心肌血流动力学功能障碍的主要介质，最近，我们证明 iPLA2γ 及其下游信号代谢物是线粒体通透性转换孔的关键调节因子，导致糖尿病心肌缺血时的坏死、坏死性凋亡和电不稳定。具体目标1，我们将使用我们生成的新型心肌细胞特异性iPLA2γ条件敲除小鼠来确定是否iPLA2γ 功能丧失可减轻糖尿病心肌中的急性缺血性损伤、电生理不稳定和脂质第二信使的适应不良生成。此外，我们证明线粒体暴露于钙离子会导致 iPLA2γ 激活，从而导致花生四烯酸 2- 的释放。花生四烯酰溶血磷脂酰胆碱，以及随后产生的多种下游生物活性脂质第二信使。将使用我们与心肌细胞特异性 iPLA2γ 功能丧失小鼠联合开发的综合质谱平台来检查 iPLA2γ 依赖性脂质第二信使产生的变化。在特定目标 2 中，我们将确定酰基辅酶 A 促进 CaMKII 的分子机制。 iPLA2ß 的磷酸化和激活将被识别、突变及其机制重要性。在特定目标 3 中，将探讨 CaMKII 介导的糖尿病心肌和缺血性糖尿病心肌中 iPLA2ß 的激活，通过甘油三酯水解、转酰基和酰基转移酶活性来催化脂质双向流动。将确定 iPLA2 参与生成第二脂质。将使用心肌细胞特异性 iPLA2ze 缺失小鼠来检查糖尿病心肌中的信使，并探讨 iPLA2ze 基因消融对糖尿病状态下心肌功能的影响。总的来说，这些研究是一种协同的多学科方法，用于确定介导糖尿病心肌病的化学机制。