Structure-Guided Analysis of Mechanisms of AT1R Functions

AT1R 功能机制的结构引导分析

基本信息

批准号：
9336426
负责人：
Sadashiva S Karnik
金额：
$ 54.8万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-15 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9336426
关键词：
Actin-Binding Protein Address Adhesions Affect Angiotensin II Angiotensin II Type 1 Receptor Blockers Angiotensin Receptor Antihypertensive Agents Atherosclerosis Binding Biology Blood Pressure Blood Vessels Cardiac Cardiovascular Physiology Cell Adhesion Cell Line Cell Survival Cell physiology Cells Cellular biology Chronic Disease Clinic Coupling Cytoskeleton Defect Development Disease Drug Targeting Endothelial Cells Equilibrium Essential Hypertension Event Functional disorder G-Protein-Coupled Receptors GTP-Binding Proteins Gene Expression Generations Goals Growth Health Heart Heart failure Homeostasis Human Hypertension Inflammatory Integrins Kidney Kidney Failure Knock-in Knowledge Laboratories Lead Ligands Link Maps Marfan Syndrome Mediating Methodology Molecular Molecular Conformation Movement Mus Mutagenesis Organ Pathology Pathway interactions Peptides Pharmaceutical Preparations Pharmacology Phosphorylation Physiological Population Property Protein Kinase PubMed Receptor Signaling Receptor, Angiotensin, Type 1 Regulation Reporting Research Risk Signal Transduction Site Site-Directed Mutagenesis Smooth Muscle Myocytes Sodium Chloride Structure Techniques Testing Tetrazoles Therapeutic Transgenic Animals Tubular formation Variant Vascular Diseases Vascular Smooth Muscle Water base blood pressure regulation carboxylate cell motility clinical efficacy clinical practice design diphenyl filamin human population study hypertension control in vivo inhibitor/antagonist insight interest migration mouse model novel novel therapeutics pleiotropism preclinical study prevent protein protein interaction receptor receptor coupling receptor function response signal processing three dimensional structure

项目摘要

DESCRIPTION (provided by applicant): The AngII type 1 receptor (AT1R) is widely known to be the master regulator of normal cardiovascular physiology. In a variety of diseases chronic stimulation of AT1R causes organ damage due to AngII-induced abnormal growth, adhesion, migration and inflammatory gene expression in cells. AT1R blockers (ARBs) effectively control hypertension but their efficacy in preventing organ damage varies widely due to unknown mechanism. Efforts have been made in several laboratories to elucidate the molecular basis of pleotropic AT1R signaling process. We have focused our research on structure, conformation and pharmacological mechanisms governing AT1R. We were the first to show ligand-independent and biased signaling in AT1R, leading to the concept of ligand modulation of subset of AT1R functions. We have recently elucidated first 3D-structure of an ARB-bound human AT1R, as an important step for beginning structure-based studies of this antihypertensive drug-target. With this new knowledge, we will address unresolved questions including: (i) how functional efficacy of clinically used drugs targeting AT1R is determined by different ligand sub-pockets within the receptor? (ii) How does a putative filamin-A binding motif embedded in AT1R operates in regulating cytoskeletal dynamics and cell adhesion properties? (iii) What aspect of activation and regulation of AT1R functions is altered by naturally occurring structural variations in AT1R? Our preliminary studies provide insight regarding (i) AT1R-ligand sub-pockets influencing differential efficacies of ARBs; (ii) AngII-induced engagement of filamin by AT1R which may be a novel pathway leading to adhesion- dependent cell functions; and (iii) possible structural effects of human AT1R variants naturally occurring in population. Our specific goals for this application are: (Aim 1) to test the hypothesis that efficacy of structuraly different ARBs in clinical practice is determined by different ligand sub-pockets found in the AT1R 3D-structure. (Aim 2) to test the hypothesis that AngII-induced AT1R interaction with filamin is regulated by a novel protein- protein interaction mechanism. (Aim 3) to test the hypothesis that AT1R variants reported in human population studies alter coupling between functional domains. We will use state-of-the-art molecular, biophysical, cell biology and in vivo techniques in our preclinical studies to advance our understanding of long unresolved issues in AT1R biology. Our findings are easily translatable to the clinic and may facilitate the development of novel therapeutics.

描述（由申请人提供）：众所周知，AngII 1 型受体（AT1R）是正常心血管生理学的主要调节剂，在多种疾病中，AT1R 的长期刺激会因 AngII 诱导的异常生长、粘附、 AT1R 阻滞剂 (ARB) 可以有效控制细胞中的迁移和炎症基因表达，但由于其机制尚不清楚，其预防器官损伤的功效差异很大。阐明多效性 AT1R 信号传导过程的分子基础我们的研究重点是控制 AT1R 的结构、构象和药理学机制我们是第一个在 AT1R 中显示配体独立和偏向信号传导的人，从而提出了配体调节子集的概念。 AT1R 功能。我们最近阐明了 ARB 结合的人类 AT1R 的第一个 3D 结构，这是开始对这种抗高血压药物靶点进行结构研究的重要一步。利用这一新知识，我们将解决尚未解决的问题，包括：（i）临床使用的靶向 AT1R 药物的功能功效如何由受体内的不同配体子口袋决定？（ii）假定的细丝蛋白 A 结合基序如何嵌入 AT1R 中？ AT1R 功能的激活和调节的哪些方面会因 AT1R 自然发生的结构变化而改变？ AT1R 配体子口袋影响 ARB 的不同功效；(ii) AngII 诱导的 AT1R 与细丝蛋白的结合，这可能是导致粘附依赖性细胞功能的新途径；以及 (iii) 人类 AT1R 变体发生的可能结构效应；我们此应用的具体目标是：（目标 1）测试以下假设：结构不同的 ARB 在临床实践中的功效是由 AT1R 中发现的不同配体子口袋决定的。 3D 结构（目标 2）检验 AngII 诱导的 AT1R 与细丝蛋白相互作用受新型蛋白质-蛋白质相互作用机制调节的假设（目标 3）检验人类研究中报告的 AT1R 变异改变两者之间耦合的假设。我们将在临床前研究中使用最先进的分子、生物物理、细胞生物学和体内技术来加深我们对 AT1R 生物学长期未解决问题的理解。可转化为临床并可能促进新疗法的开发。