Development of a gene therapy approach to treat acute lung injury using a preclinical, large animal model

使用临床前大型动物模型开发治疗急性肺损伤的基因治疗方法

基本信息

批准号：
9044084
负责人：
David A Dean
金额：
$ 78.63万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-02-01 至 2020-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9044084
关键词：
Accounting Acute Lung Injury Address Adult Adult Respiratory Distress Syndrome Affect Alveolar Alveolar wall Animal Model Animals Blood Blood Circulation Cessation of life Chest Chronic Clinical Clinical Trials Complex Data Development Disease Drops Edema Electroporation Endothelium Epithelial Failure Family suidae Future Gene Delivery Gene Expression Gene Transfer Gene therapy trial Genes Goals Grant Histology Hour Human Inflammation Inflammatory Injury Ions Ischemia Kidney Failure Length of Stay Life Liquid substance Lung Mediating Methods Microscopic Modeling Mus Myocardial dysfunction Na(+)-K(+)-Exchanging ATPase Outcome Pathologic Patients Physiologic pulse Plasmids Pre-Clinical Model Procedures Proteins Pulmonary Edema Renal function Reperfusion Therapy Resolution Respiratory physiology Sepsis Sepsis Syndrome Septic Shock Serum Sodium Channel Syndrome Testing Tight Junctions Time Tissues Translating Trauma Treatment Efficacy Water Work alveolar epithelium clinically relevant disorder prevention effective therapy electric field epithelial Na+ channel gene therapy hemodynamics improved improved outcome in vivo injured lung injury mortality neonatal patient neonatal respiratory distress non-viral gene therapy novel strategies overexpression pre-clinical preclinical study public health relevance research study respiratory distress syndrome response standard of care treatment planning

项目摘要

DESCRIPTION (provided by applicant): Acute lung injury (ALI), Acute Respiratory Distress Syndrome (ARDS), and Neonatal Respiratory Distress Syndrome (NRDS) are common, devastating clinical syndromes that affect large numbers of adult and neonatal patients (200,000 cases in the US per year) and have approximately 25% mortality with the current standard of care. We have developed a highly effective treatment for this disease in pig models that uses the ubiquitous overexpression of the Na+, K+-ATPase and epithelial sodium channel ENaC to increase alveolar fluid clearance from the previously injured lung. Our experiments show that this treatment not only improves edema resolution (and lung function and survival), but also improves alveolar epithelial/endothelial barrier function by upregulating tight junction complexes in both animal models. Highly efficient and safe gene delivery is carried out using electroporation, the application of brief synchronized square wave electric pulses across the chest. The procedure causes no trauma, no inflammation, no lung injury, no cardiac dysfunction, and uses less than 0.1 J/kg of energy. We also have developed a chronic (48 h) sepsis + gut ischemia/reperfusion pig model that accurately parallels the pathologic progression from injury to systemic inflammatory response syndrome (SIRS), to septic shock and finally to ARDS seen in human patients. Following injury, the animals are maintained, anesthetized, according to the clinical standard of care ARDSnet treatment paradigm, making comparisons to existing human clinical trial data more relevant and clear. Four hours after injury, empty control or Na+, K+-ATPase- and ENaC-expressing plasmids were electroporated into the lungs of these animals. While pigs receiving empty plasmids died from lung failure, kidney failure, and hemodynamic collapse between 24 and 40 hours after injury, animals receiving Na+,K+-ATPase- and ENaC-expressing plasmids showed greatly improved lung function, improved kidney function, less injured lungs upon gross and microscopic histological analysis, less pulmonary edema, and 60% survival (p<0.01). More impressively, an animal that received treatment plasmids when blood oxygenation dropped to the clinically defined values for ARDS of PaO2/FiO2≤300 (26 hours after injury) also showed improved lung function, survival to 48 hrs. and less injured lungs by histology. The experiments in this proposal will address questions and collect critical preclinical data needed to proceed to an IND filing with the FDA and move this treatment platform and this specific therapy forward to clinical trials. Our Aims are to (1) Test whether gene transfer of Na+, K+-ATPase alone can lessen injury and improve outcome in our pig ARDS model compared to co-transfer of Na+, K+-ATPase and ENaC genes, (2) Determine the "golden hour" or window of electroporation-mediated Na+, K+-ATPase/ENaC gene therapy treatment following injury, and (3) Determine how long the electroporation-mediated treatment provides survival and clinical benefit.

描述（由适用提供）：急性肺损伤（ALI），急性呼吸窘迫综合征（ARDS）和新生儿呼吸窘迫综合征（NRDS）很常见，毁灭性的临床综合征会影响大量的成人和新生儿患者（美国每年为200,000例），并且具有大约25％的死亡率，并具有当前标准的标准级别。我们已经在使用Na+，K+-ATPase和上皮钠通道ENAC的无处不在的猪模型中为该疾病开发了一种高效的治疗方法，以增加先前受伤的肺部肺泡液清除率。我们的实验表明，这种治疗不仅可以改善水肿的分辨率（以及肺功能和生存），而且还可以通过上调这两种动物模型中的紧密连接络合物来改善肺泡上皮/内皮屏障功能。使用电子载量进行高效且安全的基因递送，即在胸部短暂同步方波脉冲的应用。该过程不会引起创伤，没有注射，没有肺损伤，没有心脏功能障碍，并且使用少于0.1 j/kg的能量。我们还开发了一种慢性（48 h）败血症 +肠缺血/再灌注猪模型，该模型与从损伤到全身性炎症反应综合征（SIRS）的病理进展与败血性休克，最终与人类患者看到的ARDS相似。受伤后，根据临床护理ARDSNET治疗范式维持动物，麻醉，使其与现有的人类临床试验数据进行比较更加相关和清晰。受伤后四个小时，空对照或Na+，K+-ATPase-和表达ENAC的质粒被电穿孔到这些动物的肺中。受伤后24至40小时在24至40小时之间，接受空质粒的猪死于肺部衰竭，肾衰竭以及血液动力学塌陷，但接受Na+，K+-ATPase-和ENAC表达质粒的动物表现出非常改善的肺功能，改善的肾功能，肾功能改善，肺部和显微镜肺部和微观的组织学分析较少受伤，较小的肺门学或60％的幸存1％（P <0.0％）（P <0.0％）（P <0.0％）。更令人印象深刻的是，当血液氧合降至PAO2/FIO2≤300（损伤后26小时）的临床定义值时，接受治疗质粒的动物也显示出改善的肺功能，生存期至48小时。通过组织学较少受伤的肺。该提案中的实验将解决问题并收集所需的关键临床前数据，以便继续使用FDA进行IND提交，并将该治疗平台以及该特定疗法转发到临床试验。我们的目的是（1）测试与Na+，K+-ATPase和ENAC基因的转移相比，单独使用Na+，K+-ATPase的基因转移是否可以减少损伤，并改善结果，（2）确定“金色小时”或电型介导的Na+Na+，K+-ATPase/ENAC基因疗法的损伤，（3和临床益处。