Exploring the roles of Gi and S1Pr1 in endothelial barrier function in vivo

探索 Gi 和 S1Pr1 在体内内皮屏障功能中的作用

基本信息

批准号：
8028009
负责人：
Lisa Diane Wilsbacher
金额：
$ 12.53万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-01-01 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8028009
关键词：
Accounting Acute Acute Lung Injury Adult Affect Asthma Atherosclerosis Automobile Driving Award Binding Biochemical Biology Blood Vessels California Cardiology Cardiovascular system Caveolae Cell Culture Techniques Cholesterol Chronic Circadian Rhythms Couples Coupling Detection Diabetic Retinopathy Disease Doctor of Medicine Doctor of Philosophy Down-Regulation Endothelial Cells Endothelium Environment Equipment Foundations Functional disorder Future G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors GTP-Binding Proteins Genetic Goals Grant Health Hemostatic function Human In Vitro Inflammation Inflammatory Inflammatory Bowel Diseases Internal Medicine Investigation K-Series Research Career Programs Laboratories Left Liquid substance Location Maintenance Manuscripts Measures Mediating Medical Membrane Membrane Protein Traffic Mentors Mentorship Microscopy Modeling Molecular Molecular Genetics Multiple Sclerosis Mus Mutant Strains Mice Pattern Permeability Pertussis Pertussis Toxin Phenocopy Physicians Physiology Plasma Preparation Prevention Protein Chemistry Receptor Activation Recovery Regulation Reporter Reporting Research Research Personnel Research Proposals Resistance Rheumatism Role San Francisco Scientist Sepsis Series Signal Transduction Sphingosine-1-Phosphate Receptor Stimulus System Techniques Testing Thrombosis Tissues Training Training Programs Universities Vascular Permeabilities Work arrestin 1 career caveolin 1 desensitization human disease in vivo innovation insight mouse model prevent research facility response skills solute symposium tool vascular bed

项目摘要

DESCRIPTION (provided by applicant): The goal of this Career Development Award is for Dr. Wilsbacher to develop the skills necessary to successfully become an independent physician-scientist at an academic center. Dr. Wilsbacher earned her M.D. and Ph.D. degrees in the Medical Scientist Training Program at Northwestern University, where she investigated genetic and molecular mechanisms that drive circadian rhythms. She trained in Internal Medicine and Cardiology at the University of California, San Francisco. Dr. Wilsbacher's current research objective is to investigate mechanisms of vascular barrier function during normal physiology and disease, and her long-term career goal is to bridge the gap between circadian physiology and endothelial function to help advance understanding of mechanisms governing cardiovascular biology and disease. The training plan includes mentorship by Dr. Shaun Coughlin, a world-class investigator of G protein- coupled receptor (GPCR) signaling in hemostasis, thrombosis, and vascular integrity who has an established record of training young investigators to independence. Additional guidance will be provided from an expert group of scientists who are leaders in GPCR signaling (Dr. Mark von Zastrow), inflammation (Dr. Donald McDonald), and protein chemistry (Dr. James Wells). Outstanding research facilities and equipment are readily available. The training plan also incorporates advanced didactic coursework that focuses on cell signaling, membrane trafficking, protein interactions, and microscopy; attendance at seminar series; participation and presentation in local and national conferences; and mentored guidance with manuscript and grant preparation. Treating or preventing inflammation requires thorough understanding of the mechanisms of endothelial barrier function. The research proposal aims to uncover signaling mechanisms that regulate vascular integrity, particularly through sphingosine-1-phosphate receptor 1 (S1Pr1) and Gi signaling. Aim 1 investigates the role of endothelial Gi and S1Pr1 signaling in barrier function. Specifically, genetic mouse models that express pertussis toxin (which inhibits Gi) in endothelium, express a pertussis-insensitive Gi in endothelium, or conditionally delete S1Pr1 in adult endothelium will be tested for changes in vascular permeability. Aim 2 uses a new GPCR activation detection system to probe when and where S1Pr1 is activated in vivo during inflammation. The new reporter system, which transcriptionally reports on GPCR activation, is an innovative tool that will be widely applicable to other GPCRs in diverse tissues. Aim 3 probes whether caveolae influence S1Pr1-Gi coupling, and whether S1Pr1 localization in caveolae affects 2arrestin-mediated internalization. Dr. Wilsbacher's Career Development Award proposal comprises a promising candidate, outstanding training environment, rigorous training plan, and exciting research proposal that includes an innovative new tool and focuses on a topic with direct relevance to human health and disease. At the completion of this Award, Dr. Wilsbacher should have the skills necessary to succeed as an independent investigator. PUBLIC HEALTH RELEVANCE: A diverse range of acute and chronic human diseases involve abnormal endothelial permeability, including asthma, acute lung injury, inflammatory bowel disease, diabetic retinopathy, rheumatic disease, multiple sclerosis, atherosclerosis, and sepsis. Treating or preventing inflammation and its sequelae requires thorough understanding of the mechanisms of endothelial barrier function. The overall goal of this proposal is to uncover signaling mechanisms regulating vascular integrity that might be utilized in the prevention or treatment of these important human conditions.

描述（由申请人提供）：该职业发展奖的目标是让 Wilsbacher 博士培养成功成为学术中心独立医师科学家所需的技能。威尔斯巴赫博士获得了医学博士学位和博士学位。她在西北大学医学科学家培训项目中获得了博士学位，在那里她研究了驱动昼夜节律的遗传和分子机制。她在旧金山加利福尼亚大学接受了内科和心脏病学培训。 Wilsbacher 博士目前的研究目标是研究正常生理和疾病期间血管屏障功能的机制，她的长期职业目标是弥合昼夜节律生理和内皮功能之间的差距，以帮助加深对心血管生物学和疾病控制机制的理解。培训计划包括 Shaun Coughlin 博士的指导，他是 G 蛋白偶联受体 (GPCR) 止血、血栓形成和血管完整性信号传导领域的世界级研究者，在培训年轻研究者独立方面拥有良好的记录。 GPCR 信号传导（Mark von Zastrow 博士）、炎症（Donald McDonald 博士）和蛋白质化学（James Wells 博士）领域的领导者科学家专家组将提供额外指导。一流的研究设施和设备一应俱全。该培训计划还包含高级教学课程，重点关注细胞信号传导、膜运输、蛋白质相互作用和显微镜学；参加系列研讨会；参加地方和国家会议并在会上发表演讲；并为手稿和资助准备提供指导。治疗或预防炎症需要彻底了解内皮屏障功能的机制。该研究计划旨在揭示调节血管完整性的信号传导机制，特别是通过 1-磷酸鞘氨醇受体 1 (S1Pr1) 和 Gi 信号传导。目标 1 研究内皮 Gi 和 S1Pr1 信号传导在屏障功能中的作用。具体而言，将测试在内皮中表达百日咳毒素（抑制 Gi）、在内皮中表达百日咳不敏感 Gi 或有条件地删除成人内皮中 S1Pr1 的基因小鼠模型的血管通透性变化。 Aim 2 使用新的 GPCR 激活检测系统来探测炎症期间 S1Pr1 在体内激活的时间和地点。新的报告系统以转录方式报告 GPCR 激活，是一种创新工具，可广泛应用于不同组织中的其他 GPCR。目标 3 探讨小窝是否影响 S1Pr1-Gi 偶联，以及小窝中 S1Pr1 的定位是否影响 2arrestin 介导的内化。威尔斯巴赫博士的职业发展奖提案包括有前途的候选人、出色的培训环境、严格的培训计划和令人兴奋的研究提案，其中包括创新的新工具，并重点关注与人类健康和疾病直接相关的主题。完成该奖项后，威尔斯巴赫博士应具备作为独立研究者取得成功所需的技能。公共卫生相关性：多种急性和慢性人类疾病都涉及内皮通透性异常，包括哮喘、急性肺损伤、炎症性肠病、糖尿病视网膜病变、风湿性疾病、多发性硬化症、动脉粥样硬化和败血症。治疗或预防炎症及其后遗症需要彻底了解内皮屏障功能的机制。该提案的总体目标是揭示调节血管完整性的信号机制，可用于预防或治疗这些重要的人类疾病。