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)是危重病患者呼吸衰竭的常见原因。它的发病率是~ 仅在美国每年就有20万例，死亡率高得令人无法接受 报告病例的住院天数为25%-40%和360万天。虽然，我们对这些机制的理解 与ALI和急性呼吸窘迫综合征(ARDS)的发病机制和解决有关 在过去的40年里，ALI/ARDS的所有现有治疗方法仍然依赖于支持性护理，而不是 有效的治疗方案可用于改善临床结果。因此，新疗法的发展 安全、有效并基于对相关机制的深入理解的ALI/ARDS战略 ALI的发病机制是有根据的。该提案旨在阐明MTOR(机械论)特定于细胞类型的作用 [以前的哺乳动物]在ALI中的雷帕霉素靶标)，并测试同时但不同的细胞特异性的效用 瞄准MTOR以控制ALI。该提案基于我们已发表和正在进行的工作，这些工作涉及 MTOR在炎症中的细胞类型特异性作用；它介导上皮细胞的炎症，而它的作用是 限制内皮细胞炎症。然而，有趣的是，MTOR信令的“净效应”导致了 肺部的致炎表型。该提案将解决以下三个相互关联的问题，但 独立，志向远大。目标1将验证MTOR通过抗炎功能限制ALI的假设 在肺内皮细胞中。这一目标的研究将确定内皮MTOR在通过以下方式调节ALI中的作用 确定调节肺内皮细胞MTOR信号在肺炎症和损伤中的作用。 目的2将探讨MTOR通过在肺泡内发挥促炎作用而促进ALI的可能性 上皮组织。目的2研究将通过监测上皮性MTOR的作用来确定其在增强ALI中的作用 调节肺泡上皮细胞MTOR信号在肺部炎症和损伤中的作用。目标3将测试 假设以特定细胞类型的方式同时但不同地靶向MTOR(在 肺内皮细胞，但在肺泡上皮中减少它)将产生更好的保护和治疗 对阿里有利。拟议的研究将使用已建立的ALI和Will小鼠模型进行 利用一种非常新的令人兴奋的方法，使用独特的细胞特异性DNA核靶向序列(DTS) 在该质粒中引导细胞特异性的核摄取和基因(shRNA或cDNA)在所需的 单元类型。这些质粒将通过电穿孔进入小鼠的肺部，从而产生高水平的基因。 在不引起炎症或对上皮或内皮细胞层造成任何细胞损伤的情况下进行表达。这个 创造性地使用细胞特异性DTS携带质粒和电穿孔将为深入了解细胞- MTOR在ALI中的特殊作用和涉及细胞特异性调节的新治疗方法的基础 为了控制阿里。