Role of endocytosis in beta-adrenergic receptors signaling in cardiomyocytes

内吞作用在心肌细胞β-肾上腺素受体信号传导中的作用

• 批准号: 8679542
• 负责人: Roshanak Irannejad
• 金额: $ 13.43万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-04 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8679542
• 关键词: Acute; Adrenergic Agents; Adrenergic Receptor; Affinity; Agonist; Award; Biology; Biosensor; California; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular Physiology; Cardiovascular system; Catecholamines; Cell membrane; Communication; Complex; Cyclic AMP; Data; Disease; Endocytosis; Endosomes; Event; Excision; Frequencies; GTP-Binding Proteins; Generations; Goals; Grant; Heart failure; Hypertension; Image; Kinetics; Laboratories; Link; Lipids; Location; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Mentors; Microscopy; Molecular Conformation; Optics; Pathway interactions; Pharmaceutical Preparations; Phase; Physiological; Play; Production; Property; Proteins; Proteomics; Receptor Cell; Receptor Signaling; Regulation; Relative (related person); Research; Research Personnel; Role; San Francisco; Series; Signal Transduction; Specificity; System; Testing; Training; Universities; Work; Writing; abstracting; adrenergic; beta-adrenergic receptor; career; desensitization; heart function; improved; insight; interest; novel; protein activation; protein complex; receptor; receptor internalization; receptor-mediated signaling; research study; response; skills; trafficking

DESCRIPTION (provided by applicant): Project Summary/Abstract ¿-adrenergic receptors (¿ARs) play a central role in controlling the strength and frequency of cardiac contraction, and elicit Gs-linked cAMP generation. Disturbances of ¿AR signaling have been implicated in many pathological conditions such as hypertension and heart failure. Upon agonist-induced activation, ¿ARs are subject to a functional desensitization and endocytic removal from the plasma membrane (PM). Once inside the cells, the receptors were previously thought to be functionally inactive. Combining sophisticated imaging platforms with recently developed conformational biosensors, Dr. Irannejad directly probed activation of ¿2AR and its cognate Gs protein and discovered that active ¿2AR-G¿s complex is not restricted to the PM and is also present at endosomes. Her long-term goal as an independent biomedical researcher is to understand the functional consequence of the ¿AR endosomal signaling in regulating heart function. With this award, Dr. Irannejad will examine the role of endocytosis in ¿ARs signaling in cardiomyocytes. This R99/R00 award will allow her to achieve the following career goal: 1) additional training in optical microscopy, 2) additional training in cardiomyocytes biology 3) develop skills to perform proteomic studies of protein complexes, 4) develop the communication, mentoring, grant-writing, and laboratory management skills necessary to become and successful, independent biomedical researcher. The mentored phase of this research will be carried out at the University of California, San Francisco under the guidance of Dr. Mark von Zastrow. During the mentored phase, Dr. Irannejad will use the conformational biosensors in cardiomyocytes to indentify compartmentalized ¿ARs signaling (Aim1). Next, she will elucidate the role of endocytosis in signaling and contraction rate mediated by ¿1AR versus ¿2AR activation in cardiomyocytes (Aim 2). Dr. Shaun Coughlin (UCSF) director of CVRI at UCSF and an expert in cardiovascular biology will serve as co-mentor for this Aim. To indentify active ¿2AR-Gs interactomes at the endosome (Aim 3), Dr. Irannejad will begin using proteomic approach. Dr. Nevan Krogan (UCSF), experts in proteomic studies of protein complexes will serve as collaborator and advisor. During the independent phase, Dr. Irannejad will continue to focus on Aim 2 and 3, characterize the dynamics of ¿ARs-Gs mediated signaling, understand the functional consequence of the ¿2AR endosomal signaling, and identifying ¿2AR-Gs interactomes at the endosome. This award will enable Dr. Irannejad to elucidate the ¿ARs compartmentalize signaling and its functional importance in regulating heart function and potentially developing new and better drugs for the treatment of pathological cardiac function.

描述(由申请人提供)：项目摘要/摘要-肾上腺素能受体(ARs)在控制心脏收缩的强度和频率方面发挥核心作用，并诱导与Gs相关的cAMP的产生。AR信号的紊乱与高血压和心力衰竭等多种病理情况有关。在激动剂诱导的激活后，AR受到功能性脱敏和内吞从质膜(PM)上的去除。一旦进入细胞，受体以前被认为是功能不活跃的。结合复杂的成像平台和最近开发的构象生物传感器，伊拉内贾德博士直接探测了2AR及其同源Gs蛋白的激活，发现活性的2AR-G？S复合体不仅存在于PM，还存在于内小体。作为一名独立的生物医学研究人员，她的长期目标是了解AR内体信号在调节心脏功能方面的功能后果。通过这一奖项，伊兰内贾德博士将研究内吞作用在心肌细胞Ars信号中的作用。这一R99/R00奖项将使她能够实现以下职业目标：1)光学显微镜方面的额外培训；2)心肌细胞生物学方面的额外培训；3)培养进行蛋白质复合体蛋白质组学研究的技能；4)培养成为成功的独立生物医学研究人员所必需的沟通、指导、拨款和实验室管理技能。这项研究的指导阶段将在加州大学旧金山分校进行，由Mark von Zastrow博士指导。在指导阶段，伊拉内贾德博士将使用心肌细胞中的构象生物传感器来识别区分的ARs信号(Aim1)。接下来，她将阐明内吞作用在心肌细胞1AR和2AR激活所介导的信号和收缩速率中的作用(目标2)。加州大学旧金山分校心血管研究所主任、心血管生物学专家肖恩·考夫林博士将担任这一目标的共同导师。为了鉴定内体上活跃的2AR-Gs相互作用(目标3)，伊兰内贾德博士将开始使用蛋白质组学方法。蛋白质复合体蛋白质组学研究专家内万·克罗根博士将担任合作者和顾问。在独立阶段，伊兰内贾德博士将继续专注于目标2和目标3，描述Ars-Gs介导的信号动力学，了解2AR内体信号的功能后果，并确定内体的2AR-Gs相互作用。这一奖项将使伊兰内贾德博士能够阐明ARS的信号分类及其在调节心脏功能和潜在开发治疗病理性心脏功能的新的更好药物方面的功能重要性。