Degradation of The Receptor for Advanced Glycation End Products (RAGE) - A Novel Mechanism in Lung Epithelial Cell Injury

晚期糖基化终末产物受体 (RAGE) 的降解——肺上皮细胞损伤的一种新机制

• 批准号: 9326410
• 负责人: John W Evankovich
• 金额: $ 6.31万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-10 至 2019-04-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9326410
• 关键词: Acute Lung Injury; Adhesions; Adult Respiratory Distress Syndrome; Advanced Glycosylation End Products; Affect; Alpha Cell; Alveolar; Award; Biochemical; Biological; Biological Markers; Biology; Cell membrane; Cell physiology; Cells; Cellular biology; Characteristics; Charge; Clinical; Critical Care; DNA; Data; Degradation Pathway; Development; Diffuse; Doctor of Philosophy; Epithelial; Epithelial Cells; Epithelium; Extracellular Matrix; Functional disorder; Half-Life; Host Defense; Hour; Hypersensitivity; Hypoxemia; Inflammation; Inflammatory; Injury; Laboratories; Ligands; Lung; Lysine; Lysosomes; Mediating; Mediator of activation protein; Medicine; Membrane; Mentors; Modeling; Molecular; Molecular Profiling; National Research Service Awards; Natural Immunity; Pathologic; Pathway interactions; Pharmacology; Phenotype; Physicians; Physiology; Plasmids; Post-Translational Protein Processing; Process; Proteins; Recruitment Activity; Research Methodology; Research Personnel; Resistance; Respiratory Failure; Science; Scientist; Serine; Serum; Severities; Signal Transduction; Site; Small Interfering RNA; Study of serum; Survivors; Time; Training; Translational Research; Ubiquitin; Ubiquitination; Universities; Work; career; cell injury; insight; lung injury; member; mortality; novel; novel strategies; overexpression; pre-clinical; programs; protein degradation; receptor; response; skills; small molecule; translational scientist; ubiquitin-protein ligase

This application is for a Ruth L. Kirschstein National Research Service Award (NRSA) entitled “Degradation of The Receptor for Advanced Glycation End Products (RAGE) by FBXO10 – A Novel Mechanism in Lung Epithelial Cell Injury”. I am a physician in pulmonary and critical care medicine at the University of Pittsburgh. I am applying for this award to acquire advanced training in molecular physiology and cell biology, as well as new training in translational research methods and regulatory science, to develop my career as a physician scientist focused on the study of Acute Lung Injury. The main objective of my proposal is to determine how a novel RAGE degradation pathway in lung epithelial cells modulates severe lung injury. RAGE is a cell membrane receptor enriched in lung epithelia, and contributes to epithelial cell injury by weakening attachment to the extracellular matrix (ECM) and amplifying inflammatory signals in response to circulating ligands encountered in excessive inflammatory conditions such as Acute Respiratory Distress Syndrome (ARDS). Our preliminary data indicate that RAGE is degraded in lung epithelial cells in a mechanism dependent on the post-translational modification of ubiquitination. Furthermore, we have preliminarily identified a novel ubiquitin-transferring subunit termed FBXO10 responsible for targeting RAGE for degradation, and also show that RAGE is degraded in response to a ligand elevated in ARDS, CpG DNA. The aims of this study are: 1.) To determine if RAGE is ubiquitinated and degraded in lung epithelial cells by the E3-Ligase subunit FBXO10, and 2.) To determine the mechanism and biologic effect of RAGE degradation in pulmonary epithelial cells. These studies will provide insight into a novel pathologic model whereby RAGE degradation, regulated by FBXO10 mediated protein ubiquitination, controls epithelial cell detachment and excessive inflammation. FBXO10 mediated ubiquitination and degradation of RAGE may contribute to the histopathologic phenotype of “diffuse alveolar damage” characteristic of ARDS by promoting epithelial cell detachment from the ECM. Thus, modulating RAGE degradation may be a novel strategy to render epithelial cells more resistant to injury and contribute to an unmet need in novel ARDS therapies. This project will provide me advanced skills in molecular physiology and cell biology, and I will be trained in translational research methodologies and regulatory science to strengthen my development into an independent investigator. My work will be completed within the Division of Pulmonary, Allergy, and Critical Care Medicine at the University of Pittsburgh, which is dedicated to the development of physician scientists. I have committed mentoring from our Division Chief Dr. Rama Mallampalli as well as a PhD cosponsor in Dr. Bill Chen. Additionally, my mentoring committee includes Dr. Janet Lee, MD – an expert in innate immunity and host defense, and Dr. Bryan McVerry – a translational scientist and member of the Acute Lung Injury Center of Excellence in charge of the clinical ALI program.

本申请是为了申请 Ruth L. Kirschstein 国家研究服务奖 (NRSA)，题为“Degradation of FBXO10 的高级糖基化终产物 (RAGE) 受体——肺中的一种新机制 上皮细胞损伤”我是一名肺科和重症监护医学的医生。 我申请这个奖项是为了获得分子生理学和细胞生物学的高级培训， 以及转化研究方法和监管科学方面的新培训，以发展我的职业生涯 专注于急性肺损伤研究的医师科学家我的提案的主要目标是 确定肺上皮细胞中新型 RAGE 降解途径如何调节严重肺 RAGE 是肺上皮中富集的细胞膜受体，通过以下方式导致上皮细胞损伤。 削弱细胞外基质 (ECM) 的附着并放大炎症信号 循环配体在过度炎症条件下遇到，例如急性呼吸窘迫 我们的初步数据表明，RAGE 在肺上皮细胞中被降解。 依赖于泛素化翻译后修饰的机制此外，我们有。 初步鉴定出一种名为 FBXO10 的新型泛素转移亚基，负责靶向 RAGE 降解，并且还表明 RAGE 响应于 ARDS、CpG DNA 中升高的配体而降解。 本研究的目的是： 1.) 确定 RAGE 是否在肺上皮细胞中被泛素化并通过 E3-连接酶亚基 FBXO10，以及 2.) 确定 RAGE 降解的机制和生物学效应 这些研究将深入了解 RAGE 的新型病理模型。 降解由 FBXO10 介导的蛋白质泛素化调节，控制上皮细胞脱离和 FBXO10 介导的 RAGE 泛素化和降解可能导致过度炎症。 ARDS通过促进上皮细胞的“弥漫性肺泡损伤”特征的组织病理学表型 因此，调节 RAGE 降解可能是一种使上皮细胞脱落的新策略。 细胞对损伤的抵抗力更强，并有助于解决新型 ARDS 疗法中未满足的需求。 为我提供分子生理学和细胞生物学方面的高级技能，我将接受转化方面的培训 研究方法论和监管科学，以加强我发展成为独立的 我的工作将在肺科、过敏科和重症监护医学科完成。 匹兹堡大学，我致力于培养医学科学家。 我们的部门主管 Rama Mallampalli 博士以及博士共同资助者 Bil​​l 博士的指导 此外，我的指导委员会包括医学博士 Janet Lee，她是先天免疫和免疫学领域的专家。 主持人辩护，以及 Bryan McVerry 博士——一位转化科学家，也是美国急性肺损伤中心的成员 卓越负责临床 ALI 项目。