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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

基本信息

批准号：
9332419
负责人：
Sarah Marie Bass
金额：
$ 11.46万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
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：
2016-08-15 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9332419
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 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 constriction experimental study functional restoration gain of function gene therapy heart function immunocytochemistry improved in vivo insight 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

项目摘要

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.

项目摘要/摘要在过去的10年里，我的科学生涯一直致力于转化性心血管研究。我的博士研究探讨了电压门控性钾通道Kv1.5作为潜在的治疗手段的作用目标是房颤。这些研究有很强的细胞生物学重点，确定了潜在的机制渠道贩运和监管，以及药物干预如何改变这些。加入的一个目标我博士后研究的科赫实验室是为了扩大我对心血管疾病的理解在活体研究背景下的进展，更多地关注对人类的治疗干预心衰(HF)。为当前提案生成的初步数据显示，氨基(N)末端 GRK2Rgs结构域(aa45-185，βarkrgs)和一个较短的N-末端 GRK2多肽(AA 45-185，βARKNT)在心肌细胞中表达后，可改变心脏的生理功能。值得注意的是，这两种多肽似乎都能阻止压力超负荷后小鼠心力衰竭的进展，但有差异对初始肥大反应的影响。该提案的K99部分将重点放在βARKrgs βARKNT具有逆转心脏损伤后左室重构的治疗作用。这些研究将首先评估βARKrgs或βARKNT基因治疗逆转的体内疗效慢性压力超负荷时，急性适应性肥大或恢复功能。此外，我还将继续在高二和博士的指导下制作小鼠心肌梗死(MI)模型。在R00期间这项提议的阶段我将使用心脏受限转基因βARKrgs和βARKnt小鼠和我的新- 开发了基因治疗载体，以确定这些肽是否可以防止心肌梗死后的不良重塑。在K99阶段，我还将使用蛋白质组学方法来确定βARKrgs的特定结合伙伴与全长β相比，以及这些结合相互作用是否在心脏损伤或激动剂刺激。对于这些研究，我将与萨利姆·梅拉利博士密切合作坦普尔的蛋白质组学研究设施，以获得关于正确执行和培训的宝贵见解和培训蛋白质组学分析的评价。在这些研究中，βArkrgs和βArknt将成为剖析 GRK2 N末端负责蛋白质相互作用的特定结构域及其所起的作用在心血管细胞信号的调节中。该项目中发现的新的蛋白质相互作用将为独立研究提供新的途径。R00阶段的一个重点将是缩小范围并追求代表心脏信号关键元件的蛋白质结合伙伴或潜在的治疗靶点改善疾病中的心脏结构和功能。通过NIH走向独立的途径提供支持 K99/R00奖将为实现这些重要的研究目标提供必要的时间和资源，继续我的个人发展，朝着我获得终身教职的总体目标前进。