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

• 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
• 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

 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（由申请人提供）：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）是常见的破坏性临床综合征，影响大量成人和新生儿患者（美国每年200，000例），在当前标准治疗下死亡率约为25%。我们已经在猪模型中开发了一种针对这种疾病的高效治疗方法，该方法使用Na+，K+-ATP酶和上皮钠通道ENaC的普遍过表达来增加肺泡液体从先前损伤的肺中的清除。我们的实验表明，这种治疗不仅改善水肿的解决（和肺功能和生存），而且还改善肺泡上皮/内皮屏障功能，通过上调紧密连接复合物在两种动物模型。高效和安全的基因递送是使用电穿孔进行的，电穿孔是在胸部应用短暂的同步方波电脉冲。该手术不会造成创伤，没有炎症，没有肺损伤，没有心脏功能障碍，并且使用不到0.1 J/kg的能量。我们还开发了一种慢性（48小时）脓毒症+肠道缺血/再灌注猪模型，该模型准确地平行于从损伤到全身炎症反应综合征（SIRS）、脓毒性休克和最终在人类患者中观察到的ARDS的病理进展。在损伤后，根据临床护理标准ARDSnet治疗范例维持、麻醉动物，使与现有人类临床试验数据的比较更加相关和清晰。损伤后4小时，将空对照或Na+，K+-ATP酶-和ENaC-表达质粒电穿孔到这些动物的肺中。虽然接受空质粒的猪在损伤后24至40小时死于肺衰竭、肾衰竭和血流动力学衰竭，但接受表达Na+，K+-ATP酶和ENaC的质粒的动物显示肺功能大大改善，肾功能改善，在大体和显微镜组织学分析中肺损伤较少，肺水肿较少，存活率为60%（p<0.01）。更令人印象深刻的是，当血氧下降到临床定义的ARDS值PaO 2/FiO 2 ≤300（损伤后26小时）时接受治疗质粒的动物也显示出肺功能改善，存活至48小时。组织学上肺损伤较轻。该提案中的实验将解决问题并收集向FDA提交IND申请所需的关键临床前数据，并将该治疗平台和该特定疗法推向临床试验。我们的目的是（1）测试与Na+，K +-ATP酶和ENaC基因的共转移相比，单独的Na+，K+-ATP酶基因转移是否可以减轻我们的猪ARDS模型中的损伤并改善结果，（2）确定损伤后电穿孔介导的Na+，K+-ATP酶/ENaC基因治疗的“黄金时间”或窗口，和（3）确定电穿孔介导的治疗提供多长时间的存活和临床益处。