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/摘要在过去的十年中，我的科学生涯一直致力于翻译心血管研究。我的博士研究调查了电压门控钾通道KV1.5作为潜在疗法的作用靶向房颤。这些研究具有强大的细胞生物学重点，确定了基本机制渠道贩运和调节以及如何通过药物干预改变这些。加入的目标我的博士后研究的科赫实验室是扩大对心血管疾病的理解在体内研究的背景下进展，更加关注人类的热干预措施心力衰竭（HF）。为当前提案生成的初步数据表明，氨基（n）末端都 GRK2（AA 45-185，βARKRGS）和较短的N末端的RGS（G蛋白信号调节剂）域在肌细胞中表达时，GRK2（AA 45-185，βARKNT）的肽可以改变心脏生理。值得注意的是，这两个宠物都在压力越过后似乎停止了小鼠的HF进展，但具有差异对最初肥厚反应的影响。该提案的K99部分将集中于βarkrgs是否是否 βARKNT可以热起作用以逆转心脏损伤后左心室（LV）重塑。这些研究将从评估βARKRG或βARKNT基因疗法的体内效率以逆转慢性压力超负荷期间的自适应肥大或恢复功能。此外，我将继续在Erhe Gao博士的指导下实践鼠心肌梗塞（MI）模型。在R00期间该提案的阶段我将使用心脏限制的转基因βarkRG和βarknt小鼠以及我的新近开发了基因疗法载体，以确定这些petides是否可以防止MI后不良重塑。在K99阶段，I还将使用蛋白质组学方法来识别βarkRGS的特定结合伴侣与全长grk2相比，体内和βarknt 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（Salim Merali）博士紧密合作 Temple的蛋白质组学研究机构，以获得适当执行的宝贵见解和培训蛋白质组学分析的评估。在这些研究中 GRK2的N末端内的特定领域负责蛋白质相互作用及其发挥的作用在调节心血管细胞信号传导中。在这个项目中发现的新型蛋白质相互作用将为独立研究提供新的途径。 R00阶段的重点是缩小和购买代表心脏信号的关键要素或潜在治疗靶标的蛋白质结合伴侣改善疾病的心脏结构和功能。通过通往独立的NIH途径的支持 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并继续我的个人发展朝着我获得终身教师职位的整体目标。