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.

该申请是针对露丝·柯希斯坦国家研究服务奖（NRSA）的题为“退化的 FBXO10的晚期糖基化末端产品（RAGE）的受体 - 肺中的一种新机制 上皮细胞损伤”。我是大学的肺和重症监护医学的身体 匹兹堡。我正在申请该奖项，以获取分子生理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 医师科学家专注于急性肺损伤的研究。我建议的主要目的是 确定肺上皮细胞中新型的愤怒降解途径如何调节严重的肺 受伤。 RAGE是一种富含肺上皮的细胞膜受体，并通过 弱化对细胞外基质（ECM）的附着，并放大炎症信号。 循环配体在过量的炎症状态下遇到的配体，例如急性呼吸窘迫 综合征（ARDS）。我们的初步数据表明，在肺上皮细胞中愤怒降解 机制取决于泛素化的翻译后修饰。此外，我们还有 初步识别一种新型的泛素转移亚基，称为FBXO10，负责针对愤怒 降解，还表明愤怒会响应ARDS，CpG DNA中升高的配体响应。 这项研究的目的是：1。）确定愤怒是否被泛素化并通过肺上皮细胞降解 e3-ligase亚基FBXO10和2。）确定愤怒降解的机理和生物学效应 肺上皮细胞。这些研究将为新的病理模型提供洞察力，从而愤怒 由FBXO10介导的蛋白泛素化调节的降解，控制上皮细胞脱离和 过度炎症。 FBXO10介导的泛素化和愤怒降解可能有助于 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 脱离ECM。这是调节愤怒退化可能是一种新型策略来呈现上皮 细胞对损伤具有更具耐药性，并促进了新型ARDS疗法的未满足需求。这个项目将 为我提供分子生理和细胞生物学的高级技能，我将接受翻译的培训 研究方法和监管科学，将我的发展加强成独立的发展 研究者。我的工作将在肺部，过敏和重症监护医学的部门内完成 匹兹堡大学致力于发展物理科学家的发展。我承诺 我们部门负责人Rama Mallampalli博士的指导以及Bill博士的博士学位者 陈。此外，我的心理委员会包括医学博士Janet Lee博士 - 先天免疫专家和 东道国防御和布莱恩·麦克弗里（Bryan McVerry）博士 - 翻译的科学家和急性肺损伤中心的成员 临床ALI计划的卓越负责人。