Ang II-Induced Hypertension: Role of PGE2 and EP4 in End Organ Damage

Ang II 诱发的高血压：PGE2 和 EP4 在终末器官损伤中的作用

基本信息

批准号：
8120658
负责人：
MARGOT CLAIRE LAPOINTE
金额：
$ 24.37万
依托单位：
HENRY FORD HEALTH SYSTEM
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8120658
关键词：
1-Phosphatidylinositol 3-Kinase 70-kDa Ribosomal Protein S6 Kinases AGTR2 gene Abbreviations Adrenergic Agents Adrenergic Receptor Adult Angiotensin II Anterior Descending Coronary Artery Anti-Inflammatory Agents Anti-inflammatory Arrestins Arts Binding Binding Sites Biological Assay Biostatistics Core Blood Pressure Brain natriuretic peptide Caliber Cardiac Cardiac Myocytes Cardiovascular system Carrier Proteins Chronic Clathrin-Coated Vesicles Co-Immunoprecipitations Confocal Microscopy Coupled Coxibs Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cyclodextrins DTR gene Data Data Analyses Development Dinoprostone Disease Disintegrins Doctor of Medicine Doctor of Philosophy Dominant-Negative Mutation Doxycycline Dynamin EP4 receptor Eicosanoids Endocrine Endocytosis Endosomes Epidermal Growth Factor Epidermal Growth Factor Receptor Epoprostenol Equilibrium Evaluation Event Family Fibrosis Functional disorder G protein coupled receptor kinase Gases Genes Growth Harvest Heart Heart Hypertrophy Heart failure Heparin Binding Hormones Human Resources Hypertension Hypertrophy In Vitro Infiltration Inflammation Inflammatory Inflammatory Response Infusion procedures Injury Interleukin-1 Interleukins Kinins Knock-out Knockout Mice Lead Left Left Ventricular Ejection Fraction Left ventricular structure Limb structure Luciferases M-Mode Echocardiography MAP Kinase Gene Measures Mechanics Mediation Membrane Metalloproteases Microscopy Mitogen Activated Protein Kinase 1 Mitogen-Activated Protein Kinases Modeling Molecular Monomeric GTP-Binding Proteins Morphology Mus Muscle Cells Mutant Strains Mice Myocardial Infarction Natriuretic Factors Natriuretic Peptides Neonatal Organ Pathway interactions Phosphorylation Phosphotransferases Principal Investigator Process Production Property Prostaglandin E Receptor Prostaglandin-Endoperoxide Synthase Prostaglandins Protein Biosynthesis Protein Kinase C Proteins Reactive Oxygen Species Receptor Activation Receptor Protein-Tyrosine Kinases Regulation Renal function Reporter Genes Research Design Research Personnel Reverse Transcriptase Polymerase Chain Reaction Role Signal Transduction Signaling Molecule Small Interfering RNA Sodium Sodium Chloride System Testing Therapeutic Effect Thick Time Transactivation Transfection Transgenic Mice Type 2 Angiotensin II Receptor Vasodilation Ventricular Water Western Blotting Withdrawal Work absorption adrenergic arteriole autocrine base cardiovascular risk factor cyclooxygenase 2 cytokine design human WFDC2 protein improved in vivo inhibitor/antagonist knockout gene mRNA Expression macrophage monodansylcadaverine mouse PGE synthase 1 mouse model mutant mutant mouse model novel overexpression paracrine prevent programs promoter quality assurance receptor release factor response src-Family Kinases tool transcription factor

项目摘要

Hypertension is a cardiovascular risk factor that can lead to ischemic injury, myocardial infarction (Ml)and heart failure. During these disease processes, the heart undergoes hypertrophy and fibrosis, referred to as cardiac remodeling. Remodeling is stimulated by mechanical factors, release of pro-inflammatory cytokines, neurohormonal agents (p-adrenergic hormones) and vasoactive hormones with trophic properties, such as angiotensin II (Ang II). These agents also regulate the genes involved in the inflammatory response, including cyclooxygenase-2 (COX-2) and the PGE2 synthase mPGES-1, resulting in substantial production of the prostanoid PGE2 by cardiac myocytes. We have shown that COX-2 is induced in the mouse heart following Ml, and 2 wk treatment with a specific COX-2 inhibitor improves cardiac function and reduces hypertrophy and fibrosis. In vitro studies using cardiac myocytes indicate that binding of PGE2 to its receptor, EP4, results in transactivation of the epidermal growth factor receptor (EGFR), activation of p42/44 MAPK and increased protein synthesis. The gene marker of hypertrophy b-type natriuretic peptide (BMP) is also regulated by PGE2. We hypothesize that PGE2 (generated by mPGES-1) and EP4 contribute to end organ damage (cardiac hypertrophy and inflammation) in response to Ang ll-induced hypertension and ischemic injury (caused by Ml) via activation of EGFR and p42/44 MAPK. In Aim I we will study the signaling molecules involved in EGFR transactivation and downstream p42/44 MAPK activation. We will also overexpress EP4 in myocytes to study its internalization via endocytosis and the contribution of endocytosis to p42/44 MAPK signaling. In Aim II we will use transient transfection of the BMP promoter and treatment with pharmacological and molecular inhibitors of signaling molecules such as Src, small GTPases and EGFR to further elucidate EP4-dependent events in myocytes. We will also study other kinases and transcription factors downstream from p42/44 MAPK that are involved in myocyte growth, including Egr-1 and GATA-4. In Aims III and IV, we will use novel mutant mouse lines to examine how cardiac myocyte-specific overexpression and deletion of EP4 and deletion of mPGES-1 modulate hypertrophy in models of Ang ll-dependent hypertension and Ml. The contribution of the COX-2 product PGE2 and its receptor EP4 to chronic cardiac pathophysiology is virtually unexplored. Our studies will use an integrative approach to study the cellular and molecular basis for the deleterious effects of the inflammatory prostanoid PGE2 and its receptor both in vitro and in novel mouse models in vivo. Abbreviations: ACM = adult cardiac myocytes; ADAM = a disintegrin and metalloproteinase protein; Ang II = angiotensin II; p2AR = p2-adrenergic receptor; BNP = B-type natriuretic peptide; COX = cyclooxygenase; EGFR = epidermal growth factor receptor; Egr-1 = early growth response factor 1; EOD = end organ damage; EP = PGE2 receptor; EP4 = PGE2 receptor type 4; Grk = G-protein coupled receptor kinase; HB-EGF = heparin- binding epidermal growth factor; IL-1 = interleukin-1p; KO or -/- = gene knockout; LAD = left anterior descending coronary artery; LV = left ventricle; Ml = myocardial infarction; mPGES-1 = inducible membrane- localized PGE2 synthase; NVM = neonatal ventricular myocytes; p42/44 MAPK = p42/44 mitogen-activated protein kinase = Erk1/2; PGE2 = prostaglandin E2; PKA = protein kinase A; ROS = reactive oxygen species; siRNA = small interfering RNA molecule. PHS 398/2590 (Rev. 09/04, Reissued4/2006) Page 236 Continuation Format Page Principal Investigator/Program Director (Last, First, Middle): LaPointe, M.C., Ph.D/Carretero, Oscar A., M.D. A.

高血压是一种心血管危险因素，可能导致缺血性损伤，心肌梗塞（ML）和心脏衰竭。在这些疾病过程中，心脏经历肥大和纤维化，称为心脏重塑。机械因素，促炎细胞因子的释放刺激了重塑，具有营养特性（例如血管紧张素II（ANG II）。这些药物还调节炎症反应涉及的基因，包括环氧合酶2（COX-2）和PGE2合酶MPGES-1，导致大量产生心肌细胞的前列腺素PGE2。我们已经表明，ML后，在小鼠心脏中诱导COX-2，用特定COX-2抑制剂进行2周处理可改善心脏功能并降低肥大和纤维化。使用心肌细胞的体外研究表明，PGE2与其受体EP4的结合导致表皮生长因子受体（EGFR）的反式激活，P42/44 MAPK的激活和增加蛋白质合成。 PGE2还调节了肥大B型亚替肽（BMP）的基因标记。我们假设PGE2（由MPGES-1生成）和EP4造成了最终器官损伤（心脏损害肥大和炎症）响应ANG LL诱导的高血压和缺血性损伤（由ML引起）通过激活EGFR和P42/44 MAPK。在目的中，我将研究信号分子参与EGFR反式激活和下游P42/44 MAPK激活。我们还将过表达EP4 心肌细胞通过内吞作用研究其内在化以及内吞作用对P42/44 MAPK的贡献信号。在AIM II中，我们将使用BMP启动子的短暂转染和药理治疗信号分子（例如SRC，小GTPases和EGFR）的分子抑制剂，以进一步阐明肌细胞中的EP4依赖性事件。我们还将研究下游的其他激酶和转录因子来自参与肌细胞生长的P42/44 MAPK，包括EGR-1和GATA-4。在AIMS III和IV中，我们将使用新型突变小鼠系来检查心肌特异性的过表达和删除 EP4和MPGES-1的缺失在ANG LL依赖性高血压和ML模型中调节肥大。这 COX-2产品PGE2及其受体EP4对慢性心脏病理生理学的贡献实际上是未探索。我们的研究将使用综合方法研究细胞和分子基础炎症前列腺素PGE2及其受体在体外和新小鼠中的有害作用体内模型。缩写：ACM =成人心肌细胞；亚当=崩解蛋白和金属蛋白酶蛋白； Ang II = 血管紧张素II； P2AR = P2-肾上腺素受体； BNP = b型纳特里尿肽； Cox =环氧合酶； egfr =表皮生长因子受体； EGR-1 =早期生长反应因子1； EOD =最终器官损坏； EP = PGE2受体； EP4 = PGE2受体4型； GRK = G蛋白偶联受体激酶； HB-EGF =肝素 - 结合表皮生长因子； IL-1 =白介素1P; KO或 - / - =基因敲除;小伙子=左前下降冠状动脉； lv =左心室； ML =心肌梗塞； mpges-1 =诱导膜 - 局部PGE2合酶； NVM =新生儿心室心肌细胞； P42/44 MAPK = P42/44促分裂原激活蛋白激酶= ERK1/2; PGE2 =前列腺素E2; PKA =蛋白激酶A; ROS =活性氧； siRNA =小干扰RNA分子。 PHS 398/2590（Rev. 09/04，重新发射4/2006）第236页延续格式页面首席调查员/计划主管（最后，第一，中间）：Lapointe，M.C。，博士/Carretero，Oscar A.，M.D。一个。