Unique Domain Features of GRK2 and Roles in Cardiovascular Disease

GRK2 的独特结构域特征及其在心血管疾病中的作用

• 批准号: 9899299
• 负责人: Sarah Marie Bass
• 金额: $ 24.9万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9899299
• 关键词: ADRBK1 gene; Acute; Agonist; American; Atrial Fibrillation; Award; Binding; Binding Proteins; Binding Sites; Cardiac; Cardiac development; Cardiovascular Diseases; Cardiovascular system; Cause of Death; Cell Surface Receptors; Cellular biology; Chronic; Complex; Coronary artery; Data; Development; Disease; Disease Progression; Elements; Evaluation; Faculty; G protein coupled receptor kinase; G-Protein-Coupled Receptors; Gene Transduction Agent; Goals; Heart; Heart Diseases; Heart Hypertrophy; Heart Injuries; Heart failure; Human; Hypertension; Hypertrophy; Immunohistochemistry; In Vitro; Infarction; Injury; Intervention; Investigation; Lead; Left; Left Ventricular Remodeling; Length; Ligation; Link; Measures; Mediating; Modeling; Molecular; Mus; Muscle Cells; Myocardial Infarction; N-terminal; Pathway interactions; Patients; Peptides; Pharmacologic Substance; Phase; Phosphorylation; Physiological; Physiology; Play; Positioning Attribute; Proteins; Proteomics; RGS Domain; Regulation; Research; Resources; Role; Signal Pathway; Signal Transduction; Structure; Techniques; Testing; Therapeutic; Therapeutic Intervention; Time; Training; Transgenic Mice; Transgenic Organisms; United States National Institutes of Health; Up-Regulation; Ventricular; Voltage-Gated Potassium Channel; Work; cardiogenesis; career; comorbidity; constriction; experimental study; functional restoration; gain of function; gene therapy; heart function; heart preservation; immunocytochemistry; improved; in vivo; in vivo evaluation; insight; ischemic injury; mouse model; new therapeutic target; novel; pressure; prevent; protein protein interaction; protein transport; receptor; reduce symptoms; research facility; response; tenure track; therapeutic development; therapeutic target; tool; trafficking; vector

PROJECT SUMMARY/ABSTRACT For the past 10 years my scientific career has been devoted to translational cardiovascular research. My doctoral studies investigated the role of the voltage-gated potassium channel Kv1.5 as a potential therapeutic target atrial fibrillation. These studies had a strong cell biology focus, determining the mechanisms underlying channel trafficking and regulation and how these were altered by pharmaceutical intervention. A goal in joining the Koch lab for my postdoctoral studies was to broaden my understanding of cardiovascular disease progression within the context of in vivo studies, with a greater focus on therapeutic interventions for human heart failure (HF). Preliminary data generated for the current proposal shows that both the amino(N)-terminal RGS (Regulator of G-protein Signaling) domain of GRK2 (aa 45-185, βARKrgs) and a shorter N-terminal peptide of GRK2 (aa 45-185, βARKnt) can alter cardiac physiology when expressed in myocytes. Of note, these two peptides both appear to halt HF progression in mice after pressure-overload but have differential effects on the initial hypertrophic response. The K99 portion of this proposal will focus on whether βARKrgs and βARKnt can act therapeutically to reverse left-ventricular (LV) remodeling after cardiac injury. These studies will begin with an evaluation of the in vivo efficacy of βARKrgs or βARKnt gene-therapy to reverse adaptive hypertrophy acutely or restore function during chronic pressure overload. In addition, I will continue to practice the murine myocardial infarction (MI) model under the guidance of Dr. Erhe Gao. During the R00 phase of this proposal I will use the cardiac-restricted transgenic βARKrgs and βARKnt mice and my newly- developed gene therapy vectors to determine whether these peptides prevent adverse remodeling post-MI. During the K99 phase I will also use proteomic approaches to identify specific binding partners for βARKrgs and βARKnt in vivo, compared to full-length GRK2, and whether these binding interactions are altered after cardiac injury or upon agonist stimulation. For these studies I will work closely with Dr. Salim Merali, Director of the Proteomics Research Facility at Temple, to gain invaluable insight and training in the proper execution and evaluation of proteomic analysis. In these studies βARKrgs and βARKnt will serve as powerful tools to dissect the specific domains within the N-terminus of GRK2 responsible for protein interactions and the role they play in the regulation of cardiovascular cell signaling. Novel protein interactions discovered in this project will provide new avenues for independent research. A focus of the R00 phase will be to narrow down and pursue the protein binding partners that represent key elements of cardiac signaling or potential therapeutic targets for improving cardiac structure and function in disease. Support through the NIH Pathway to Independent K99/R00 award would provide the necessary time and resources for achieving these important research goals, and continuing my personal development toward my overall goal of obtaining a tenure-track faculty position.

项目总结/文摘