microRNA miR-147 Dampens Alveolar Epithelial Inflammation During ARDS

microRNA miR-147 抑制 ARDS 期间的肺泡上皮炎症

• 批准号: 10316251
• 负责人: Holger K. Eltzschig
• 金额: $ 45.88万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10316251
• 关键词: 3' Untranslated Regions; Acute Lung Injury; Acute Respiratory Distress Syndrome; Acute respiratory failure; Alveolar; Applications Grants; Attenuated; Binding; Binding Sites; Biological Models; Breeding; Clinical Trials; DNA Sequence Alteration; Epithelial Cells; Funding; Generations; Genes; Genetic; Genetic study; Goals; HIF1A gene; Homozygote; Human; Hypoxia; In Vitro; Inflammation; Inflammatory; Link; Liquid substance; Lung; Mechanical ventilation; Mechanics; Mediating; Messenger RNA; MicroRNAs; Microarray Analysis; Modeling; Mosaicism; Mucositis; Mucous Membrane; Mus; Mutation; Operative Surgical Procedures; Pathway interactions; Patients; Periodicity; Pulmonary Edema; Pulmonary Inflammation; Repression; Resolution; Resuscitation; Role; Stretching; Supportive care; Testing; Therapeutic; Tissues; Toll-like receptors; Untranslated Regions; Ventilator-induced lung injury; alveolar epithelium; base; design; experience; in vitro Model; in vivo; lung injury; macrophage; nanoparticle; neutrophil; non cardiogenic pulmonary edema; novel therapeutic intervention; overexpression; preclinical safety; pressure; prevent; promoter; safety study; therapeutic miRNA; transcription factor; treatment strategy; ventilation

Project Summary The main goal of this grant application is to identify microRNA (miRNA) targets that protect the alveolar epithelium from excessive inflammation during acute lung injury (ALI). ALI is characterized by acute respiratory failure in the setting of non-cardiogenic pulmonary edema. It causes acute respiratory distress syndrome (ARDS) in humans. The current application is focused on identifying miRNAs that could be targeted to dampen alveolar epithelial inflammation. To identify miRNA targets that dampen alveolar inflammation, we exposed mice to ventilator-induced lung injury (VILI) and isolated alveolar epithelial cells. A microarray analysis identified miR-147 as a leading candidate. Confirmatory studies demonstrated that miR-147 is induced during ALI by mechanical ventilation in vivo or through in vitro exposure of alveolar epithelia to cyclic mechanical stretch. In vivo elevations of miR-147 levels during ALI persisted despite depletion of neutrophils or macrophages. In addition, we identified a role for hypoxia-inducible transcription factor HIF1A for miR-147 induction. miR-147-/- mice experience more severe lung injury during VILI. A search for miR-147 targets identified toll-like receptor adaptor molecule 2 (TICAM2) as pro- inflammatory target. In line with a link of miR-147 with TICAM2 expression, we found that alveolar epithelia isolated from miR-147loxp/loxp SPC Cre+ mice showed elevated Ticam2 levels. Similarly, increased pulmonary edema during VILI of miR-147loxp/loxp SPC Cre+ mice was resuscitated by concomitant genetic deletion of Ticam2. miR-147 overexpression via miR-147-containing nano-particles was protective during ALI. Finally, proof-of-principle studies in ARDS patients showed elevated miR-147 levels in their BAL fluid. Therefore, we hypothesize that HIF-dependent induction of miR-147 represents an endogenous pathway to dampen lung inflammation. We designed 3 specific aims, where we will first study the expression of miR-147 utilizing in vitro modeling systems (Aim 1). Aim 2 is focused on in vivo studies of ALI, where we will utilize tissue-specific approaches of miR-147 deletion and examine mice with a mutation of the miR-147 binding site in the TICAM2 3' untranslated region (genetic targeting for these mice was successful). Finally, we will target miR-147 for the treatment of ARDS in Aim 3.

项目摘要 该赠款应用的主要目标是识别保护肺泡的microRNA（miRNA）目标 急性肺损伤（ALI）期间过度炎症的上皮。 ALI的特征是急性呼吸道 在非心脏病肺水肿的情况下发生故障。它导致急性呼吸窘迫综合征（ARDS） 在人类中。当前的应用集中在识别可以降低肺泡的miRNA 上皮炎症。 为了确定抑制肺泡炎症的miRNA靶标，我们将小鼠暴露于呼吸机诱导的肺 损伤（VILI）和分离的牙槽上皮细胞。微阵列分析将miR-147确定为领先的候选人。 验证性研究表明，通过体内机械通气在ALI期间诱导miR-147 通过体外暴露于肺泡上皮症中的环状机械拉伸。 miR-147水平的体内升高 尽管中性粒细胞或巨噬细胞耗尽，但在阿里期间仍持续存在。此外，我们确定了 缺氧诱导转录因子HIF1A用于miR-147诱导。 mir-147 - / - 小鼠经历更严重的肺 Vili期间受伤。搜索miR-147靶标确定了类似收费的受体适配器分子2（TICAM2） 炎症目标。与miR-147与TICAM2表达的链接一致，我们发现肺泡上皮 从miR-147loxp/loxp spc cre+小鼠中分离出来的TICAM2水平升高。同样，增加 miR-147loxp/loxp spc cre+小鼠在VILI期间的肺水肿通过同时遗传复苏 TICAM2的删除。在ALI期间，通过含MiR-147的纳米颗粒的miR-147过表达具有保护性。 最后，对ARDS患者的原则研究表明，其BAL液中miR-147水平升高。所以， 我们假设MiR-147的HIF依赖性诱导代表了潮湿肺的内源性途径 炎。我们设计了3个特定目标，我们将首先研究使用体外的miR-147的表达 建模系统（AIM 1）。 AIM 2专注于ALI的体内研究，我们将利用组织特异性 miR-147缺失的方法，并检查小鼠用miR-147结合位点突变在tiCAM2 3'中 未翻译的区域（这些小鼠的遗传靶向成功）。最后，我们将以Mir-147为目标 AIM 3中的ARDS处理。