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

• 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
• 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

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）：这个职业发展奖的目标是博士Wilsbacher发展必要的技能，成功地成为一个独立的医生，科学家在学术中心。威尔斯巴赫博士获得了医学博士学位和博士她在西北大学的医学科学家培训项目中获得学位，在那里她研究了驱动昼夜节律的遗传和分子机制。她在加州大学旧金山分校弗朗西斯科接受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生理和内皮功能之间的差距，以帮助促进对心血管生物学和疾病机制的理解。 培训计划包括Shaun Coughlin博士的指导，Shaun Coughlin博士是止血、血栓形成和血管完整性中G蛋白偶联受体（GPCR）信号传导的世界级研究者，他在培训年轻研究者独立性方面有着良好的记录。GPCR信号传导（Mark von Zastrow博士）、炎症（Donald McDonald博士）和蛋白质化学（James威尔斯博士）领域的领导者科学家专家组将提供额外的指导。优秀的研究设施和设备是现成的。培训计划还包括先进的教学课程，重点是细胞信号，膜贩运，蛋白质相互作用和显微镜;出席研讨会系列;参与和介绍地方和国家会议;和指导指导与手稿和赠款准备。 治疗或预防炎症需要彻底了解内皮屏障功能的机制。该研究提案旨在揭示调节血管完整性的信号机制，特别是通过鞘氨醇-1-磷酸受体1（S1 Pr 1）和Gi信号传导。目的1研究内皮细胞Gi和S1 Pr 1信号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（其抑制Gi）、在内皮中表达百日咳不敏感Gi或在成人内皮中条件性缺失S1 Pr 1的遗传小鼠模型的血管通透性变化。目的2使用一种新的GPCR激活检测系统来探测S1 Pr 1在炎症过程中何时何地在体内被激活。新的报告系统，转录报告GPCR激活，是一种创新的工具，将广泛适用于其他GPCR在不同的组织。目的3探讨小窝是否影响S1 Pr 1-Gi偶联，以及S1 Pr 1在小窝中的定位是否影响2arrestin介导的内化。 Wilsbacher博士的职业发展奖提案包括一个有前途的候选人，出色的培训环境，严格的培训计划，以及令人兴奋的研究提案，其中包括一个创新的新工具，并专注于与人类健康和疾病直接相关的主题。在这个奖项完成后，Wilsbacher博士应该有必要的技能，成功地作为一个独立的研究者。 公共卫生相关性：多种急性和慢性人类疾病涉及异常内皮渗透性，包括哮喘、急性肺损伤、炎性肠病、糖尿病视网膜病变、风湿性疾病、多发性硬化、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传导机制，这些机制可能用于预防或治疗这些重要的人类疾病。