Mitigating Acute Lung Injury by Cell-specific Targeting of MTOR

通过细胞特异性靶向 MTOR 减轻急性肺损伤

• 批准号: 10187645
• 负责人: David A Dean
• 金额: $ 58.94万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10187645
• 关键词: Acute; Acute Lung Injury; Address; Adult Respiratory Distress Syndrome; Alveolar Cell; Animal Model; Anti-Inflammatory Agents; Blood Vessels; Bronchopulmonary Dysplasia; CMV promoter; Case Study; Cell Nucleus; Cells; Cellular Metabolic Process; Cessation of life; Chronic Obstructive Airway Disease; Clinical; Complementary DNA; Critical Illness; DNA; DNA Polymerase III; Data; Development; Disease; Drug Targeting; Effectiveness; Electroporation; Endothelial Cells; Endothelium; Epithelial; Epithelial Cells; FRAP1 gene; Functional disorder; Gases; Gene Delivery; Gene Expression; Gene Transfer; Genes; In Vitro; Incidence; Infiltration; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Injury; Knock-out; Knockout Mice; Lead; Length of Stay; Lung; Lung Inflammation; Mediating; Modeling; Monitor; Mus; Nuclear; Outcome; Pathogenesis; Patients; Phenotype; Plasmids; Preventive; Public Health; Publishing; Pulmonary Edema; Reporting; Resolution; Respiratory Failure; Role; Sepsis; Signal Transduction; Sirolimus; Structure; Supportive care; Testing; Therapeutic; Tumor Angiogenesis; United States; Work; alveolar epithelium; base; cDNA Expression; cecal ligation puncture; cell growth; cell injury; cell type; design; effective therapy; effectiveness evaluation; experimental study; gene therapy; improved; improved outcome; innovation; insight; knock-down; lung development; lung injury; mortality; mouse model; novel; novel strategies; novel therapeutic intervention; prevent; promoter; response; small hairpin RNA; therapeutically effective; transgene expression; treatment strategy; uptake; vascular injury

Acute lung Injury (ALI) is a common cause of respiratory failure in critically ill patients. It has an incidence of ~ 200,000 cases each year in the United States alone and is associated with an unacceptably high mortality rate of 25-40% and 3.6 million hospital days in reported cases. Although, our understanding of the mechanisms relevant to the pathogenesis and the resolution of ALI and Acute Respiratory Distress Syndrome (ARDS) has increased during the past four decades, all current therapies for ALI/ARDS still rely on supportive care and no effective therapeutic options are available to improve clinical outcome. Thus, the development of new treatment strategies for ALI/ARDS that are safe, effective, and based on deeper understanding of the mechanisms involved in ALI pathogenesis is warranted. This proposal aims to clarify the cell type-specific role of MTOR (mechanistic [formerly mammalian] target of rapamycin) in ALI and test the utility of simultaneous but differential cell-specific targeting of MTOR to control ALI. The proposal is based on our published and on-going work that implicates a cell type-specific role for MTOR in inflammation; it mediates inflammation in epithelial cells whereas it serves to limit endothelial cell inflammation. Intriguingly, however, the “net effect” of MTOR signaling results in a proinflammatory phenotype in the lung. The proposal will address the following three inter-related, but independent, aims. Aim 1 will test the hypothesis that MTOR limits ALI by serving an anti-inflammatory function in pulmonary endothelium. Studies in this aim will ascertain the role of endothelial MTOR in moderating ALI by determining the effects of modulating MTOR signaling in pulmonary endothelium on lung inflammation and injury. Aim 2 will investigate the possibility that MTOR promotes ALI by exerting a proinflammatory function in alveolar epithelium. Aim 2 studies will determine the role of epithelial MTOR in augmenting ALI by monitoring the effects of modulating MTOR signaling in alveolar epithelium on lung inflammation and injury. Aim 3 will test the hypothesis that targeting MTOR simultaneously but differentially in a cell type-specific manner (increasing it in pulmonary endothelium but decreasing it in alveolar epithelium) will yield a superior protective and therapeutic benefit against ALI. The proposed studies will be carried out using established mouse models of ALI and will utilize a very new and exciting approach that uses unique cell-specific DNA nuclear targeting sequences (DTSs) in the plasmid to direct cell-specific plasmid nuclear uptake and gene (shRNA or cDNA) expression in the desired cell type. The plasmids will be delivered into the lungs of mice via electroporation, which yields high level of gene expression without inducing inflammation or any cell damage to the epithelial or endothelial cell layer. The creative use of cell-specific DTS carrying plasmid and electroporation will provide valuable insight into the cell- specific role of MTOR in ALI and the basis for novel therapeutic approaches involving cell-specific modulation of MTOR to control ALI.

急性肺损伤（ALI）是重症患者呼吸衰竭的常见原因。它有一个事件〜 仅在美国，每年每年有200,000例案件，并且与高死亡率相关 报告病例的25-40％和360万日期。虽然，我们对机制的理解 与ALI和急性呼吸窘迫综合征（ARDS）的发病机理和分辨率有关 在过去的四十年中，所有目前的ALI/ARD疗法都依赖于支持性护理，没有 有效的治疗选择可改善临床结果。那，发展新待遇 安全，有效且基于对所涉及机制的更深入了解的ALI/ARD的策略 在ALI发病机理中有必要。该建议旨在阐明MTOR（机械）特定细胞类型的作用 [以前为哺乳动物]雷帕霉素的靶标）在ALI中并测试简单但差异细胞特异性的实用性 靶向MTOR控制ALI。该提案是基于我们发表且正在进行的工作，该工作实现了 MTOR在炎症中的细胞类型特异性作用；它介导上皮细胞的注射 限制内皮细胞炎症。然而，有趣的是，mTOR信号的“净效应”导致A 肺中促炎的表型。该提案将解决以下三个相关的问题，但 独立，目标。 AIM 1将测试MTOR通过抗炎功能限制ALI的假设 在肺化学室中。以此目的的研究将确定佛教室MTOR在主持Ali中的作用 确定调节MTOR信号在肺部内皮中对肺部感染和损伤的影响。 AIM 2将通过在肺泡中发挥促炎功能来调查MTOR促进ALI的可能性 上皮。 AIM 2研究将通过监测效果来确定上皮MTOR在增强ALI中的作用 在肺部感染和损伤中调节MTOR信号传导。 AIM 3将测试 假设仅以细胞类型的方式靶向MTOR但以不同的方式靶向（增加） 肺部内皮但在肺泡上皮下降低）将产生优越的保护和治疗 对阿里的利益。提出的研究将使用已建立的ALI小鼠模型进行，将 利用一种非常新颖而令人兴奋的方法，该方法使用独特的细胞特异性DNA核靶向序列（DTSS） 在质粒中直接在所需的细胞特异性质粒核摄取和基因（shRNA或cDNA）表达中 细胞类型。质粒将通过电穿孔传递到小鼠的肺部，该电穿孔产生高水平的基因 无诱导注射或对上皮细胞层或任何细胞损害的表达。 携带质粒和电穿孔的细胞特异性DT的创造性使用将为细胞提供宝贵的见解。 MTOR在ALI中的具体作用，新型治疗方法的基础涉及细胞特异性调节 控制阿里的mtor。