Central GRK5 modulation of Angiotensin II receptor expression in heart failure

GRK5 对心力衰竭中血管紧张素 II 受体表达的中枢调节

• 批准号: 8531707
• 负责人: Karla Haack
• 金额: $ 5.39万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8531707
• 关键词: Address; Anesthesia procedures; Angiotensin II; Angiotensin II Receptor; Animal Model; Animals; Attenuated; Baroreflex; Brain; Brain Stem; Brain region; Cardiac; Cause of Death; Cell Line; Cells; Chronic; Complex; Coronary artery; Data; Diagnosis; Disease; Dissociation; Dominant-Negative Mutation; Echocardiography; Excretory function; Exercise; Frequencies; G protein coupled receptor kinase; G-Protein-Coupled Receptors; GRK; GRK5 gene; Genetic Transcription; Heart Rate; Heart failure; Hypertension; Hypothalamic structure; In Vitro; Injection of therapeutic agent; Laboratories; Lead; Ligation; Link; Measures; Mediating; Mus; Muscle; Nerve; Neurons; Norepinephrine; Nuclear; Patients; Phosphorylation; Phosphotransferases; Plasma; Plasmids; Play; Process; Quality of life; Rattus; Receptor, Angiotensin, Type 1; Recycling; Regulation; Role; Small Interfering RNA; Subfamily lentivirinae; Techniques; Testing; Therapeutic; Training; Transcriptional Regulation; Ubiquitin-Proteasomal Pathway; United States; Up-Regulation; effective therapy; hemodynamics; improved; indexing; overexpression; paraventricular nucleus; pressure; prevent; protein expression; receptor; receptor expression; receptor upregulation; research study; response; sedentary; therapeutic development; therapeutic target; urinary

DESCRIPTION (provided by applicant): This proposal will address the following specific aims: (1) Determine the role(s) of central GRK5 in the regulation of AT1R (Angiotensin II type 1 Receptor) expression under normal conditions and during Chronic Heart Failure (CHF). (2) Identify the mechanism(s) of modulation of central AT1R and GRK5 following Exercise Training (ExT) in CHF animals. (3) Determine the role of cytosolic and nuclear GRK5 in the transcriptional regulation of AT1R by I?B¿ and NF-?B. In Specific Aim 1 we will induce CHF by coronary artery ligation. Overexpression of GRK5 will be targeted to the RVLM or PVN by lentiviral injection. To determine the effects of GRK5 knockdown on AT1R expression and changes in sympatho-excitation, we will utilize both commercially available GRK5 KO mice and lentiviral packaged siRNA against GRK5 that will be injected into the RVLM or PVN. Urinary excretion of norepinephrine (NE), plasma NE and Ang II will be measured in all animal groups. Arterial pressure and heart rate will be continuously recorded in order to derive additional indices of sympatho-excitation and to determine arterial baroreflex function. Sympathetic nerve activity will be directly recorded under anesthesia in terminal experiments. In all animals cardiac function will be evaluated serially by high-frequency echocardiography. In Specific Aim 2, we will induce CHF in GRK5KO mice that are either sedentary or ExT as well as utilize CHF rats in which GRK5 has been silenced in the PVN or RVLM using siRNA lentivirus. Following ExT, sympathetic and baroreflex function will be evaluated in a similar fashion as in Specific Aim 1. We will test Specific Aim 3 in CATH.a neurons, utilizing both overexpression and silencing techniques with a GRK5 plasmid and siRNA to examine the subcellular localization of AT1R, I?B¿, NF-?B, and GRK5 following Ang II stimulation. We will also perform parallel studies using a K215R dominant negative GRK5 construct to determine if the GRK5/AT1R/I?B¿/NF-?B interaction is kinase-dependent.

描述（由申请人提供）：本提案将解决以下具体目标：（1）确定正常条件下和慢性心力衰竭（CHF）期间中枢GRK 5在调节AT 1 R（血管紧张素II 1型受体）表达中的作用。 (2)确定CHF动物运动训练（ExT）后中枢AT 1 R和GRK 5的调节机制。 (3)确定的作用，胞浆和核GRK 5在转录调节AT 1 R的I？B和NF-？B。在特定目标1中，我们将通过冠状动脉结扎诱导CHF。GRK 5的过表达将通过慢病毒注射靶向RVLM或PVN。为了确定GRK 5敲低对AT 1 R表达和交感神经兴奋变化的影响，我们将利用市售GRK 5 KO小鼠和将注射到RVLM或PVN中的针对GRK 5的慢病毒包装siRNA。将在所有动物组中测量去甲肾上腺素（NE）、血浆NE和Ang II的尿排泄。将连续记录动脉压和心率，以获得交感神经兴奋的其他指数并确定动脉压力反射功能。在终末实验中，将在麻醉下直接记录交感神经活动。在所有动物中，心脏 将通过高频超声心动图连续评价功能。在特定目标2中，我们将在久坐或ExT的GRK 5 KO小鼠中诱导CHF，以及利用其中GRK 5已使用siRNA慢病毒在PVN或RVLM中沉默的CHF大鼠。ExT后，将以与特定目标1相似的方式评价交感神经和压力反射功能。我们将测试具体目标3在CATH。a神经元，利用过表达和沉默技术与GRK 5质粒和siRNA检查AT 1 R，I？B，NF-？B和GRK 5。我们还将进行平行研究，使用K215 R显性阴性GRK 5构建，以确定是否GRK 5/AT 1 R/I？B/NF-？B相互作用依赖于激酶。