Endothelin and Angiotensin Receptor Signaling in Pulmonary Hypertension

肺动脉高压中的内皮素和血管紧张素受体信号传导

• 批准号: 7993574
• 负责人: Peter Rudolph Polgar
• 金额: $ 49.64万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-02-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7993574
• 关键词: Angiotensin Receptor; Angiotensins; Attenuated; Blood Vessels; Cell Proliferation; Cell membrane; Cells; Clinical Research; Contracts; Cyclic AMP; Cyclic GMP; Disease; Elements; Endothelial Cells; Endothelin; Endothelin A Receptor; Endothelin Receptor; Endothelin-1; Epoprostenol; Equilibrium; Etiology; G-substrate; Generations; Growth; Health; Heterotrimeric GTP-Binding Proteins; Homeostasis; Human; Hydrolysis; Iloprost; Individual; JAK2 gene; Lead; Learning; Lung; MAP Kinase Gene; MAPK14 gene; MAPK8 gene; Measurement; Measures; Membrane; Monocrotaline; Monomeric GTP-Binding Proteins; Muscle Contraction; Mutation; Nitric Oxide; PTEN gene; Participant; Patients; Peptide Receptor; Peptides; Pharmaceutical Preparations; Phospholipase A2; Phosphorylation; Physiological; Play; Production; Prostaglandins I; Pulmonary Hypertension; Rattus; Receptor Cell; Receptor Signaling; Regulation; Respiratory physiology; Role; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Staging; System; TBC 11251; Testing; Therapeutic; Transfection; Transplantation; Vascular Endothelial Cell; Vascular Endothelium; Vascular remodeling; Vasodilator Agents; Work; autocrine; bosentan; constriction; design; follow-up; mutant; novel strategies; paracrine; receptor; receptor function; research study; sildenafil; transmission process; vasoconstriction

DESCRIPTION (provided by applicant):Drugs of choice used in the treatment of pulmonary hypertension (PH) clearly show that endothelin-1 (ET-1) is a major participant in this fatal disease. Angiotensin (AngII) through its AT1 receptor stimulates the production of ET-1 in human lung vascular endothelial cells (HLVEC). ET-1 then stimulates the production of vasodilating, antiproliferative nitric oxide (NO) and prostacyclin (PGI2) through its ETB receptor in the HLVEC. In human lung vascular smooth muscle cells (HLVSMC) ET-1 stimulates cell proliferation and vasoconstriction. In PH this homeostatic regulation breaks down. The AT1 receptor, which is involved at the initial stage of PH, propagates many of the signals participating in the etiology of PH. Our studies on receptor signaling motifs combined with mutant receptor transfection and signal cascade determinations have enabled us to control the signaling of the AT1 receptor. More recently we can now control signaling by the (wild type) endogenously expressed receptors with the use of cell penetrating peptides constructed to mimic the receptor motifs sequence(s). We intend to achieve signal control in the ETA and ETB receptors and then regulate these signal transductions in human lung vascular endothelial and smooth muscle cells. It is anticipated that this approach will lead to new treatments of PH. Our working hypothesis is that signal cascades involving MAPKs, the Akt/PI3K/PTEN system, small G-protein RhoA and heterotrimeric G- proteins work in various combinations to produce ET-1 through the AT1 receptor and to produce NO and PGI2 through the ETB receptor in human lung vascular endothelial cells and to produce contraction and proliferation through the ETA/ETB receptors in human lung vascular smooth muscle cells; that these signal interactions can be controlled through the use of altered receptors and cell penetrating peptides; and that by altering these signals, we will be able to decrease ET-1 and increase NO and prostacyclin production by endothelial cells and decrease growth and contraction in smooth muscle cells. Specific aims, 1) a) We will regulate AT1, ETA and ETB receptor signaling through motif identification and mutation construction within the receptor to regulate G1q, G1i, RhoA, Akt and MAPKs. b) We will then regulate signaling in endogenously expressed AT1 and ETB receptors in human lung endothelial cells and ETA receptors in smooth muscle cells with use of membrane permeable peptides. 2) With use of the altered receptors and the peptides developed in SA 1, we will regulate the expression and release of ET-1 by AngII and the release of NO and prostacyclin in the HLVEC by ET-1. These results will then be used to control function of these receptors in HLVEC isolated from lung transplants of individuals with PH. 3) With use of the altered receptors and the peptides developed in Specific Aim 1, we will regulate contraction and proliferation in human lung vascular smooth muscle cells, including cells obtained from individuals with PH. 4) The studies on smooth muscle contraction in culture will be followed up with physiological experiments using rats. PUBLIC HEALTH RELEVANCE: Pulmonary hypertension is a debilitating and mostly fatal disease. Our experiments, including the production of peptides which interfere with negative signals of angiotensin and endothelin, are designed to lead to new approaches which should prove beneficial in the treatment of pulmonary hypertension.

描述（由申请人提供）：用于治疗肺动脉高压（PH）的首选药物清楚地表明内皮素-1（ET-1）是这种致命疾病的主要参与者。血管紧张素 (AngII) 通过其 AT1 受体刺激人肺血管内皮细胞 (HLVEC) 产生 ET-1。然后，ET-1 通过 HLVEC 中的 ETB 受体刺激血管舒张、抗增殖的一氧化氮 (NO) 和前列环素 (PGI2) 的产生。在人肺血管平滑肌细胞 (HLVSMC) 中，ET-1 刺激细胞增殖和血管收缩。在PH中，这种体内平衡调节被破坏。 AT1 受体参与 PH 的初始阶段，传播许多参与 PH 病因的信号。我们对受体信号传导基序的研究与突变受体转染和信号级联测定相结合，使我们能够控制 AT1 受体的信号传导。最近，我们现在可以使用模拟受体基序序列的细胞穿透肽来控制（野生型）内源表达受体的信号传导。我们打算实现ETA和ETB受体的信号控制，然后调节人肺血管内皮细胞和平滑肌细胞中的这些信号转导。预计这种方法将带来PH的新治疗方法。我们的工作假设是，涉及 MAPK、Akt/PI3K/PTEN 系统、小 G 蛋白 RhoA 和异源三聚体 G 蛋白的信号级联以各种组合方式发挥作用，通过 AT1 受体产生 ET-1，通过人肺血管内皮细胞中的 ETB 受体产生 NO 和 PGI2，并通过人肺血管内皮细胞中的 ETA/ETB 受体产生收缩和增殖。 平滑肌细胞；这些信号相互作用可以通过使用改变的受体和细胞穿透肽来控制；通过改变这些信号，我们将能够减少 ET-1，增加内皮细胞 NO 和前列环素的产生，并减少平滑肌细胞的生长和收缩。具体目标，1) a) 我们将通过受体内的基序识别和突变构建来调节 AT1、ETA 和 ETB 受体信号传导，从而调节 G1q、G1i、RhoA、Akt 和 MAPK。 b) 然后，我们将使用膜渗透肽来调节人肺内皮细胞中内源表达的 AT1 和 ETB 受体以及平滑肌细胞中 ETA 受体的信号传导。 2)利用SA 1中开发的改变的受体和肽，我们将调节AngII对ET-1的表达和释放以及ET-1对HLVEC中NO和前列环素的释放。然后，这些结果将用于控制从 PH 患者肺移植中分离出的 HLVEC 中这些受体的功能。 3) 通过使用特定目标 1 中开发的改变的受体和肽，我们将调节人肺血管平滑肌细胞的收缩和增殖，包括从 PH 个体获得的细胞。 4) 培养物中平滑肌收缩的研究将通过大鼠的生理实验来进行。公共卫生相关性：肺动脉高压是一种使人衰弱且大多致命的疾病。我们的实验，包括干扰血管紧张素和内皮素负信号的肽的生产，旨在产生新的方法，这些方法应该被证明对治疗肺动脉高压有益。