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: 10539334
• 负责人: John W Evankovich
• 金额: $ 16.12万
• 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/10539334
• 关键词: Acute Lung Injury; Acute Respiratory Distress Syndrome; Authorization documentation; 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; Endosomes; 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; inflammatory modulation; 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.

本申请适用于 K08 指导临床科学家研究职业发展奖，名为 “RNF216 响应血浆 MicroRNA 降解 Toll 样受体 8——一种新机制 调节急性肺损伤中的炎症”。我是一名肺科和重症监护医学的医生 匹兹堡大学。我申请该奖项是为了获得细胞生物学、转化学方面的高级培训 研究方法和生物信息学来发展我作为一名专注于研究的医师科学家的职业生涯 急性呼吸窘迫综合征（ARDS）。我提案的主要目标是确定一部小说如何 单核细胞中的 Toll 样受体 8 (TLR8) 降解途径可调节严重肺损伤。 TLR8是一种模式 识别受体，可感知免疫原性 RNA，包括一些源自宿主的血浆 microRNA。 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 TLR8 在单核细胞中以某种机制降解 依赖于泛素化的翻译后修饰。此外，我已经确定了一个候选泛素- 转移称为 RNF216 的 E3 连接酶负责靶向 TLR8 进行降解，我观察到 ARDS 患者中 RNF216 mRNA 表达降低。最后，我确定了循环的一个子集 ARDS 受试者血浆 microRNA 可能充当新型 TLR8 配体。本研究的目的是：i) 定义单核细胞中泛素/蛋白酶体系统调节 TLR8 蛋白水平的机制 ii) 确定 RNF216 是否通过指导 TLR8 降解来调节炎症信号传导并定义 RNF216 ARDS 中的表达，以及 iii) 检查 ARDS 受试者中血浆 miRNA 作为 TLR8 配体。这些研究将 深入了解一种新的病理生物学模型，其中 TLR8 降解由 RNF216 介导 蛋白质泛素化，控制 ARDS 中对宿主血浆 microRNA 的炎症反应。等离子体 miRNA 诱导的 TLR8 激活，通过减少 RNF216 介导的泛素化和降解而增强 TLR8，可能会导致过度炎症。因此，调节 TLR8 降解可能是减少 TLR8 降解的新策略。 ARDS 中的过度炎症反应。该项目将为我提供细胞生物学方面的高级技能 分析分子数据集的生物信息方法。我将接受转化研究方法方面的培训 加强我成为一名独立调查员的发展。我已承诺接受我们部门的指导 首席教授 Rama Mallampalli 博士和博士导师 Bill Chen 博士。此外，我的指导委员会 其中包括先天免疫领域的国际专家 Robert Lafyatis 博士、免疫领域权威专家 Stephen Chan 博士 翻译 miRNA 生物学，以及 Bryan McVerry 博士——一位出色的转化科学家和 Acute 成员 肺损伤卓越中心负责监督临床急性肺损伤项目。我的工作即将完成 匹兹堡大学肺科、过敏科和重症监护医学科 致力于医师科学家的发展。