B-Arrestins and G Protein-Coupled Receptor Kinases in Cardiovascular Function

B-抑制蛋白和 G 蛋白偶联受体激酶对心血管功能的影响

• 批准号: 8098814
• 负责人: ROBERT J LEFKOWITZ
• 金额: $ 39万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8098814
• 关键词: Acute; Adrenergic Receptor; Agonist; Angioplasty; Angiotensin II; Angiotensins; Arrestins; Binding; Biological Assay; Blood Pressure; Blood Vessels; Bypass; Calsequestrin; Cardiac; Cardiac Myocytes; Cardiovascular Physiology; Cardiovascular system; Catecholamines; Cells; Chemotaxis; Chronic; Development; Fluorescence Resonance Energy Transfer; G protein coupled receptor kinase; G-Protein-Coupled Receptors; GTP-Binding Proteins; Generations; Heart; Heart failure; Hyperplasia; Hypertension; In Vitro; Individual; Kinetics; Knockout Mice; Ligands; MAP Kinase Modules; Mediating; Mitogen-Activated Protein Kinases; Mitogens; Modeling; Mus; Operative Surgical Procedures; Pathway interactions; Phosphotransferases; Physiological; Play; Primary Myocardial Diseases; Protein Isoforms; Receptor Activation; Regulation; Role; Second Messenger Systems; Signal Transduction; Small Interfering RNA; Smooth Muscle Myocytes; Specificity; Stimulation of Cell Proliferation; System; Testing; Transgenic Mice; Vasodilation; analog; arrestin B; attenuation; cell growth; desensitization; in vivo; migration; mutant; novel; novel therapeutics; overexpression; receptor; receptor-mediated signaling; research study; scaffold; second messenger; vascular smooth muscle cell migration; vasoconstriction

DESCRIPTION (provided by applicant): P-ARRESTINS AND GRKs IN CARDIOVASCULAR FUNCTION. G protein coupled receptors (GPCRs) such as those for catecholamines and angiotensin II regulate cardiovascular functions including vasoconstriction and vasodilation; vascular smooth muscle cell (VSM) mitogenesis and migration; and cardiac inotropy and chronotropy. These functions may be perturbed in hypertension, in pathological intimal hyperplasia as after bypass surgery or angioplasty, or in heart failure. Following GPCR activation, the receptors are phosphorylated by one of several G protein coupled receptor kinases (GRKs) and then bind one of the two isoforms of p-arrestin. p-arrestin binding sterically interdicts further signaling to G proteins leading to receptor desensitization and attenuation of second messenger generation, p-arrestins also play positive roles in signaling, serving as adaptors and scaffolds to organize receptor-mediated activation of MAP kinase cascades and other pathways. These MAP kinases regulate mitogenesis and migration of vascular smooth muscle cells. Recently we have found that receptor ligands may vary dramatically in their ability to stimulate G protein versus p-arrestin-mediated signaling. However, the cellular and physiological consequences of p-arrestin/GRK mediated signaling in cardiovascular (or other) systems are largely unknown. Accordingly, we will use several approaches to test our central hypothesis that p-arrestin/GRK- mediated signaling contributes significantly to the short and long-term regulation of cardiovascular function. Our approaches will include the use of mutant receptors and specific agonists which can selectively activate p-arrestin but not G protein signaling; siRNA to p-arrestins and GRKs; knock-out mice which we have previously developed lacking each of the p-arrestins and GRKs; and cells derived there from. Utilizing the p-adrenergic receptors and angiotensin IMA receptor as our models our specific aims are to determine: 1) the specificity of individual p-arrestins and GRKs in desensitizing G protein mediated second messenger generation and to compare this with their roles in mediating signaling, as for example through ERK activation. 2) The physiological effects of P-arrestin-mediated signaling, a) In vitro on cardiac myocytes and vascular smooth muscle cells, and b) In vivo on cardiac inotropy and chronotropy and on systemic blood pressure, c) The in vivo consequences of cardiac signaling by mutant p-adrenergic receptors and angiotensin II1A receptors which are uncoupled from G proteins but signal through p-arrestins. 3) The chronic effects of an ang analog (SII ang) which signals only through p-arrestins, on the development and course of CHF in a transgenic mouse overexpressing calsequestrin in the heart. These experiments have the potential to delineate an entirely novel and general mode of receptor-mediated signaling and to point the way to the development of novel therapeutics which target this recently discovered signaling mechanism.

描述（由申请人提供）：P-ARRESSTIN 和 GRK 在心血管功能中的作用。 G 蛋白偶联受体 (GPCR)，例如儿茶酚胺和血管紧张素 II，可调节心血管功能，包括血管收缩和血管舒张；血管平滑肌细胞（VSM）有丝分裂和迁移；以及心肌正性肌力和变时性。这些功能可能在高血压、搭桥手术或血管成形术后的病理性内膜增生或心力衰竭中受到干扰。 GPCR 激活后，受体被几种 G 蛋白偶联受体激酶 (GRK) 之一磷酸化，然后结合 p-arrestin 的两种亚型之一。 p-arrestin 结合在空间上阻止进一步向 G 蛋白发出信号，导致受体脱敏和第二信使生成减弱，p-arrestin 在信号传导中也发挥积极作用，充当接头和支架来组织受体介导的 MAP 激酶级联和其他途径的激活。这些 MAP 激酶调节血管平滑肌细胞的有丝分裂和迁移。最近我们发现受体配体刺激 G 蛋白的能力与 p-抑制蛋白介导的信号传导的能力可能存在显着差异。然而，p-arrestin/GRK 介导的心血管（或其他）系统信号传导的细胞和生理后果在很大程度上尚不清楚。因此，我们将使用多种方法来检验我们的中心假设，即 p-arrestin/GRK 介导的信号传导对心血管功能的短期和长期调节有显着贡献。我们的方法将包括使用突变受体和特异性激动剂，它们可以选择性激活 p-arrestin，但不能激活 G 蛋白信号传导； p-抑制蛋白和 GRK 的 siRNA；我们之前开发的缺乏 p-arrestins 和 GRK 的基因敲除小鼠；以及从中衍生的细胞。利用 p-肾上腺素受体和血管紧张素 IMA 受体作为模型，我们的具体目标是确定：1​​) 各个 p-抑制蛋白和 GRK 在使 G 蛋白介导的第二信使生成脱敏中的特异性，并将其与它们在介导信号传导中的作用（例如通过 ERK 激活）进行比较。 2) P-抑制蛋白介导的信号转导的生理效应，a) 体外对心肌细胞和血管平滑肌细胞的影响，b) 体内对心肌正性肌力和变时性以及全身血压的影响，c) 突变型 p-肾上腺素受体和血管紧张素 II1A 受体对心脏信号转导的体内影响，这些受体与 G 蛋白解偶联，但通过信号传递 p-抑制蛋白。 3) 仅通过β-抑制蛋白发出信号的血管紧张素类似物（SII 血管紧张素）对心脏中过度表达calsequestrin 的转基因小鼠的CHF 的发展和病程的慢性影响。这些实验有可能描绘出一种全新且通用的受体介导信号传导模式，并为开发针对这种最近发现的信号传导机制的新型疗法指明了道路。

项目成果

Distinct phosphorylation sites on the β(2)-adrenergic receptor establish a barcode that encodes differential functions of β-arrestin.

• DOI: 10.1126/scisignal.2001707
• 发表时间: 2011-08-09
• 期刊: Science signaling
• 影响因子: 7.3
• 作者: Nobles KN;Xiao K;Ahn S;Shukla AK;Lam CM;Rajagopal S;Strachan RT;Huang TY;Bressler EA;Hara MR;Shenoy SK;Gygi SP;Lefkowitz RJ
• 通讯作者: Lefkowitz RJ

Therapeutic potential of β-arrestin- and G protein-biased agonists.

• DOI: 10.1016/j.molmed.2010.11.004
• 发表时间: 2011-03
• 期刊: Trends in molecular medicine
• 影响因子: 13.6
• 作者: Whalen EJ;Rajagopal S;Lefkowitz RJ
• 通讯作者: Lefkowitz RJ

β-Arrestin regulation of myosin light chain phosphorylation promotes AT1aR-mediated cell contraction and migration.

• DOI: 10.1371/journal.pone.0080532
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Simard E;Kovacs JJ;Miller WE;Kim J;Grandbois M;Lefkowitz RJ
• 通讯作者: Lefkowitz RJ

Emerging paradigms of β-arrestin-dependent seven transmembrane receptor signaling.

• DOI: 10.1016/j.tibs.2011.06.003
• 发表时间: 2011-09
• 期刊: TRENDS IN BIOCHEMICAL SCIENCES
• 影响因子: 13.8
• 作者: Shukla, Arun K.;Xiao, Kunhong;Lefkowitz, Robert J.
• 通讯作者: Lefkowitz, Robert J.

Multiple ligand-specific conformations of the β2-adrenergic receptor.

• DOI: 10.1038/nchembio.634
• 发表时间: 2011-08-21
• 期刊: NATURE CHEMICAL BIOLOGY
• 影响因子: 14.8
• 作者: Kahsai, Alem W.;Xiao, Kunhong;Rajagopal, Sudarshan;Ahn, Seungkirl;Shukla, Arun K.;Sun, Jinpeng;Oas, Terrence G.;Lefkowitz, Robert J.
• 通讯作者: Lefkowitz, Robert J.