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、破坏性的临床综合征，影响大量成人和新生儿患者（200,000 例）美国每年），按照目前的护理标准，死亡率约为 25%。我们已经在猪模型中开发了一种高效的治疗方法，该方法使用Na+、K+-ATP 酶和上皮钠通道 ENaC 的普遍过度表达可增加先前受伤肺部的肺泡液清除率。我们的实验表明，这种治疗不仅可以改善水肿消退（以及肺功能和存活率），还可以改善肺泡上皮/细胞。使用电穿孔（短暂同步方波的应用）通过上调紧密连接复合物来实现内皮屏障功能。该手术不会造成创伤、无炎症、无肺损伤、无心脏功能障碍，并且使用的能量低于 0.1 J/kg。我们还发现了慢性（48 小时）脓毒症 + 肠道缺血/再灌注。猪模型准确地模拟了人类患者从损伤到全身炎症反应综合征（SIRS）、感染性休克和最终 ARDS 的病理进展，根据临床标准对动物进行维持和麻醉。 care ARDSnet 治疗范例，使与现有人类临床试验数据的比较更加相关和清晰。受伤后四小时，将空对照或表达 Na+、K+-ATPase 和 ENaC 的质粒电穿孔到这些动物的肺部。损伤后 24 至 40 小时内，质粒因肺衰竭、肾衰竭和血流动力学崩溃而死亡，接受 Na+、K+-ATPase 和 ENaC 表达质粒的动物表现出极大的死亡。肺功能改善，肾功能改善，肉眼和显微镜组织学分析中肺部损伤减少，肺水肿减少，存活率达到 60%（p<0.01），更令人印象深刻的是，当血氧饱和度降至临床定义值时，接受治疗质粒的动物。对于 PaO2/FiO2 ≤ 300（损伤后 26 小时）的 ARDS，组织学结果显示肺功能有所改善，存活时间可达 48 小时，并且肺部损伤较少。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进临床试验。我们的目标是 (1) 测试是否仅进行 Na+、K+-ATP 酶的基因转移。与 Na+、K+-ATPase 和 ENaC 基因共转移相比，可以减少猪 ARDS 模型中的损伤并改善结果，(2) 确定电穿孔介导的“黄金时间”或窗口损伤后进行 Na+、K+-ATPase/ENaC 基因治疗，以及 (3) 确定电穿孔介导的治疗可提供多长时间的生存和临床益处。