Degradation of Toll-Like Receptor 8 by RNF216 in response to plasma MicroRNAâs â A Novel Mechanism Regulating Inflammation in Acute Lung Injury

RNF216 响应血浆 MicroRNA 降解 Toll 样受体 8 – 调节急性肺损伤炎症的新机制

• 批准号: 10338104
• 负责人: John W Evankovich
• 金额: $ 16.16万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-15 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10338104
• 关键词: Acute Lung Injury; Acute Respiratory Distress Syndrome; Award; Bioinformatics; Biology; Cell physiology; Cellular biology; Cessation of life; Characteristics; Clinical; Critical Care; Cytoplasmic Tail; Data; Data Set; Degradation Pathway; Development; Doctor of Philosophy; Family; Gene Expression; Gene Proteins; Hypersensitivity; Infection; Inflammation; Inflammatory; Inflammatory Response; International; Invaded; K-Series Research Career Programs; Ligands; Lung; Lysine; Mediating; Medicine; Mentors; MicroRNAs; Modeling; Molecular; Molecular Profiling; Natural Immunity; Organ failure; Pathologic; Pathway interactions; Patients; Pattern; Pattern recognition receptor; Phenotype; Physicians; Plasma; Post-Translational Protein Processing; Process; Proteins; RNA; Research; Research Methodology; Research Personnel; Respiratory Failure; Scientist; Signal Pathway; Signal Transduction; Site; Source; Survivors; System; TLR8 gene; Training; Translational Research; Ubiquitin; Ubiquitination; Universities; Work; authority; career; cohort; cytokine; immune activation; immunogenic; insight; lung injury; mRNA Expression; member; molecular modeling; monocyte; mortality; multicatalytic endopeptidase complex; novel; novel strategies; pathogen; programs; protein degradation; protein expression; recruit; response; skills; translational scientist; ubiquitin-protein ligase

This application is for a K08 Mentored Clinical Scientist Research Career Development Award entitled “Degradation of Toll-Like Receptor 8 by RNF216 in response to plasma MicroRNA’s – A Novel Mechanism Regulating Inflammation in Acute Lung 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 cell biology, translational research methods, and bioinformatics to develop my career as a physician scientist focused on the study of acute respiratory distress syndrome (ARDS). The main objective of my proposal is to determine how a novel Toll-Like Receptor 8 (TLR8) degradation pathway in monocytes regulates severe lung injury. TLR8 is a pattern recognition receptor that senses immunogenic RNA, including some host-derived plasma microRNA’s. TLR8 activation initiates signaling leading to the secretion of cytokines, contributing to excessive inflammation that is characteristic of severe ARDS. My preliminary data indicate that TLR8 is degraded in monocytes in a mechanism dependent on the post-translational modification of ubiquitination. Further, I have identified a candidate ubiquitin- transferring E3 ligase termed RNF216 responsible for targeting TLR8 for degradation, and I observe that RNF216 mRNA expression is decreased in patients with ARDS. Lastly, I have identified a subset of circulating plasma microRNA’s in ARDS subjects that may function as novel TLR8 ligands. The aims of this study are: i) to define the mechanism regulating TLR8 protein levels by the ubiquitin/proteasome system in monocytes ii) to determine if RNF216 modulates inflammatory signaling by directing TLR8 degradation and define RNF216 expression in ARDS, and iii) to examine plasma miRNA’s in ARDS subjects as TLR8 ligands. These studies will provide insight into a novel pathobiologic model whereby TLR8 degradation, regulated by RNF216 mediated protein ubiquitination, controls inflammation in response to host-derived plasma microRNA’s in ARDS. Plasma miRNA-induced TLR8 activation, augmented by reduced RNF216 mediated ubiquitination and degradation of TLR8, may drive excessive inflammation. Thus, modulating TLR8 degradation may be a novel strategy to reduce excessive inflammatory responses in ARDS. This project will provide me advanced skills in cell biology and bioinformatic approaches to analyze molecular datasets. I will be trained in translational research methodologies to strengthen my development into an independent investigator. I have committed mentoring from our Division Chief, Dr. Rama Mallampalli and a PhD comentor in Dr. Bill Chen. Additionally, my mentoring committee includes Dr. Robert Lafyatis – an international expert in innate immunity, Dr. Stephen Chan – an authority in translational miRNA biology, and Dr. Bryan McVerry – a superb translational scientist and member of the Acute Lung Injury Center of Excellence overseeing the clinical Acute Lung Injury program. My work will be completed within the Division of Pulmonary, Allergy, and Critical Care Medicine at the University of Pittsburgh, which is committed to the development of physician scientists.

本申请是为K 08指导临床科学家研究职业发展奖， “RNF 216对Toll样受体8的降解反应--一种新的机制” 急性肺损伤中的炎症调节”。我是一名肺部和重症监护医学医师， 匹兹堡大学我申请这个奖项是为了获得细胞生物学的高级培训， 研究方法和生物信息学，以发展我作为一名医生科学家的职业生涯，专注于研究 急性呼吸窘迫综合征（ARDS）。我的建议的主要目的是确定一部小说 单核细胞中Toll样受体8（TLR 8）降解途径调节严重肺损伤。TLR 8是一种 免疫原性RNA是一种识别受体，其感测免疫原性RNA，包括一些宿主来源的血浆微小RNA。TLR8 激活启动导致细胞因子分泌的信号传导，导致过度炎症， 严重ARDS的特征。我的初步数据表明，TLR 8在单核细胞中的降解机制 依赖于遍在蛋白化的翻译后修饰。另外，我发现了一种候选的泛素- 转移E3连接酶RNF 216负责靶向TLR 8降解，我观察到， ARDS患者RNF 216 mRNA表达降低。最后，我已经确定了一个循环的子集， 可能作为新型TLR 8配体发挥作用的血浆microRNA。本研究的目的是：（一） 定义单核细胞中泛素/蛋白酶体系统调节TLR 8蛋白水平的机制ii） 确定RNF 216是否通过指导TLR 8降解来调节炎症信号传导，并定义RNF 216 iii）检测ARDS受试者中作为TLR 8配体的血浆miRNA。这些研究将 提供了对新型病理生物学模型的深入了解，其中TLR 8降解由RNF 216介导调节 蛋白质泛素化，控制炎症反应的主机来源的血浆microRNA的在ARDS。血浆 miRNA诱导的TLR 8激活，通过减少RNF 216介导的泛素化和降解 TLR 8可能导致过度炎症。因此，调节TLR 8降解可能是一种新的策略，以减少 过度的炎症反应。这个项目将为我提供先进的细胞生物学技能， 生物信息学方法来分析分子数据集。我将接受转化研究方法的培训 来帮助我成长为一名独立调查员我已经承诺从我们的司指导 主任，拉马马兰帕利博士和比尔陈博士的博士评论员。另外，我的指导委员会 包括先天免疫领域的国际专家Robert Lafyatis博士， 翻译miRNA生物学，和Bryan McVerry博士-一位出色的翻译科学家和Acute 肺损伤卓越中心监督临床急性肺损伤计划。我的工作将完成 在匹兹堡大学的肺、过敏和重症监护医学部， 致力于医学科学家的发展。