Electroporation-mediated Pulmonary Gene Transfer for Acute Lung Injury

电穿孔介导的肺基因转移治疗急性肺损伤

• 批准号: 8070439
• 负责人: David A Dean
• 金额: $ 71.27万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8070439
• 关键词: Acute; Acute Lung Injury; Adenovirus Vector; Adenoviruses; Adult; Adult Respiratory Distress Syndrome; Aerosols; Affect; Alveolar; Animal Model; Animal Testing; Animals; Biological Assay; Blood Chemical Analysis; Bolus Infusion; Bronchoscopes; Cell Count; Cessation of life; Chest; Clinical; Clinical Trials; DNA delivery; Disease; Drug Formulations; Electrodes; Electroporation; Family suidae; Future; Gene Combinations; Gene Delivery; Gene Expression; Gene Transfer; Genes; Goals; Histology; Human; Inflammatory Response; Injury; Irrigation; Laboratories; Length of Stay; Ligation; Lipids; Liquid substance; Lung; Mediating; Methodology; Methods; Modality; Modeling; Movement; Mus; Na(+)-K(+)-Exchanging ATPase; Neonatal; Newborn Respiratory Distress Syndrome; Non-Viral Vector; Patients; Phase; Physiologic pulse; Plasmids; Polymers; Positioning Attribute; Pre-Clinical Model; Proteins; Pulmonary Edema; Puncture procedure; Rattus; Recombinants; Recommendation; Reporter Genes; Research; Safety; Saline; Sepsis; Series; Severities; Side; Structure of parenchyma of lung; Surface; Syndrome; Techniques; Technology; Testing; Translating; Trauma; Treatment Efficacy; Tube; United States; Viral; Viral Vector; Work; alveolar epithelium; base; clinical application; cytokine; design; efficacy testing; electric field; endotracheal; epithelial Na+ channel; gene therapy; human disease; in vivo; injured; lung injury; mortality; mouse model; non-viral gene delivery; non-viral gene therapy; novel strategies; plasmid DNA; prevent; research study; response; standard of care; success; surfactant; therapeutic gene; vector; viral vector development

DESCRIPTION (provided by applicant): PROJECT SUMMARY/ABSTRACT 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 up to 40% mortality with the current standard of care. Our ultimate goal is to develop nonviral gene therapy approaches to treat ALI. We have shown that electroporation can be used to transfer genes to lungs with established lung injury and pulmonary edema to treat the disease and lessen the severity of the injury. The method is simple, fast, and safe. In order to move this toward clinical application, we must demonstrate its efficacy and safety in a large animal model that represents the human disease and develop the most simple and safe method possible in order for it to be accepted by clinicians. The highest recommendation for an animal model is good evidence that the results of the study would be similar in a clinical trial To this end, we have recently applied this approach to a group pigs (35-40 kg) using energies less than 0.1 J/kg and have achieved gene transfer to the lungs with no mortality or injury. Our R21 Phase specific aims are to (1) optimize electroporation parameters for safety and gene transfer to the lungs of pigs, (2) evaluate methods of DNA delivery, (3) develop and optimize a bronchoscope-based electrode for lung gene delivery, and (4) determine whether gene transfer of ENaC as well as Na,K-ATPase subunit genes increases efficacy of gene therapy for treatment of lung injury in a mouse model. At the end of the R21 phase, we will have established optimal parameters for gene delivery to the pig lung and have determined the best therapeutic gene combination to treat lung injury in two complementary pig models of ALI. The R33 phase specific aims are (1) evaluate efficacy of electroporation-mediated gene transfer of Na,K-ATPase/ENaC gene subunits to prevent lung injury in a saline lavage model of mild-moderate ALI, (2) determine whethere transfer of these genes can treat pre-existing lung injury in this model, (3) determine whether transfer of these genes can protect from lung injury in a severe sepsis-induced cecal ligation and pucture model of ARDS, and (4) evaluate whether this approach can be used to treat previously established sepsis-induced lung injury in this model. These studies will provide us with the proof-of-principle for transthoracic pulmonary electroporation and establish the safety and efficacy in an two established pre-clinical models. PROJECT NARRATIVE Acute lung injury, Acute Respiratory Distress Syndrome (ARDS), and Neonatal Respiratory Distress Syndrome are common, devastating clinical syndromes that affect an estimated 200,000 adult and neonatal patients each year in the US and have up to 40% mortality with the current standard of care. We have developed a nonviral gene therapy approach using electric fields that can prevent as well as treat existing ARDS in two small animal models. The work proposed in this study will extend these findings to a large animal model to evaluate whether these findings can be translated into larger subjects and ultimately humans. At the end of the study, we will have demonstrated efficacy of this novel approach to treat lung injury in an established pre-clinical model.

描述（由申请人提供）：项目摘要/摘要急性肺损伤（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，毁灭性的临床综合征，这些临床综合症会影响大量的成人和新生儿患者（每年200,000例）（每年200,000例），并具有多达40％的标准标准的抵押次数多达40％。我们的最终目标是开发非病毒基因治疗方法来治疗ALI。我们已经表明，电穿孔可用于将基因转移到肺部损伤和肺水肿的肺部，以治疗疾病并减轻损伤的严重程度。该方法简单，快速且安全。为了将其转向临床应用，我们必须在代表人类疾病的大型动物模型中证明其功效和安全性，并开发了最简单，最安全的方法，以便临床医生接受它。对动物模型的最高建议是很好的证据表明，在此目的的临床试验中，该研究的结果将相似，我们最近使用小于0.1 J/kg的能量将这种方法应用于猪（35-40 kg），并将基因转移到肺部没有死亡率或伤害。 Our R21 Phase specific aims are to (1) optimize electroporation parameters for safety and gene transfer to the lungs of pigs, (2) evaluate methods of DNA delivery, (3) develop and optimize a bronchoscope-based electrode for lung gene delivery, and (4) determine whether gene transfer of ENaC as well as Na,K-ATPase subunit genes increases efficacy of gene therapy for treatment of lung injury in a mouse 模型。在R21阶段结束时，我们将建立最佳的基因递送到猪肺的参数，并确定了在两个互补的ALI互补猪模型中治疗肺损伤的最佳治疗基因组合。 R33相的特异性目的是（1）评估电穿孔介导的基因转移的基因转移的功效，K-ATPase/ENAC基因基因亚基在盐水灌洗模型中预防肺损伤，（（2）确定这些基因的盐分转移是否可以在此模型中进行预疾病，确定这些基因的损伤 - 3）是否可以在这些模型中进行损伤，（3）（3）（3）（3）（3）（3）（3）（3）（3）（3）（3）（3）（3） ARDS的连接和触发模型，以及（4）评估该方法是否可用于治疗此模型中先前确定的败血症诱导的肺损伤。这些研究将为我们提供经胸腔肺电穿孔的原则证明，并在两个已建立的临床前模型中建立安全性和功效。项目叙事急性肺损伤，急性呼吸窘迫综合征（ARDS）和新生儿呼吸窘迫综合征是常见的，毁灭性的临床综合征会影响美国估计的200,000名成人和新生儿患者，并且与当前的护理标准有关40％的死亡率。我们已经使用电场开发了一种非病毒基因治疗方法，该方法可以预防和治疗两个小动物模型中的现有ARDS。这项研究中提出的工作将把这些发现扩展到大型动物模型，以评估这些发现是否可以转化为较大的受试者和最终人类。在研究结束时，我们将在既定的临床前模型中证明这种新型方法治疗肺损伤的功效。