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: 7494800
• 负责人: David A Dean
• 金额: $ 38.5万
• 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/7494800
• 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; Caring; Cell Count; Cessation of life; Chest; Clinical; Clinical Trials; Condition; 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; Numbers; Patients; Phase; Physiologic pulse; Plasmids; Polymers; Positioning Attribute; Pre-Clinical Model; Proteins; Pulmonary Edema; Pulse taking; Puncture procedure; Rate; Rattus; Recombinants; Recommendation; Reporter; Research; Respiratory distress; Safety; Saline; Sepsis; Series; Severities; Side; Standards of Weights and Measures; Structure of parenchyma of lung; Surface; Sus scrofa; Syndrome; Techniques; Technology; Testing; Translating; Trauma; Treatment Efficacy; Tube; United States; Viral; Viral Vector; Work; abstracting; alveolar epithelium; base; clinical application; cytokine; design; 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; 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.

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，影响了大量成人和新生儿患者（美国每年20万例），在目前的护理标准下，死亡率高达40%。我们的最终目标是开发非病毒基因治疗方法来治疗ALI。我们已经证明，电穿孔可用于将基因转移到已建立的肺损伤和肺水肿的肺部，以治疗疾病并减轻损伤的严重程度。该方法简便、快速、安全。为了将其推向临床应用，我们必须在代表人类疾病的大型动物模型中证明其有效性和安全性，并开发出最简单、最安全的方法，以便临床医生接受。为了达到这个目的，我们最近将这种方法应用于一群猪（35-40公斤），使用低于0.1 J/kg的能量，并实现了基因转移到肺部，没有死亡或损伤。我们的R21期特异性目标是：(1)优化安全性和基因转移到猪肺的电穿孔参数，(2)评估DNA传递方法，(3)开发和优化基于支气管镜的肺基因传递电极，(4)确定ENaC以及Na， k - atp酶亚基基因的基因转移是否能提高基因治疗小鼠肺损伤的疗效。在R21期结束时，我们将确定基因传递到猪肺的最佳参数，并确定两种互补性ALI猪模型中治疗肺损伤的最佳治疗基因组合。R33期特异性目的是(1)评估电穿孔介导的Na、k - atp酶/ENaC基因亚基基因转移在轻度-中度ALI生理盐水灌洗模型中预防肺损伤的效果，(2)确定这些基因的转移是否可以治疗该模型中已有的肺损伤，(3)确定这些基因的转移是否可以保护严重脓毒症诱导的ARDS盲肠结扎和穿刺模型中的肺损伤。(4)评估该方法是否可用于治疗先前建立的脓毒症肺损伤模型。这些研究将为我们提供经胸肺电穿孔的原理证明，并在两个已建立的临床前模型中建立安全性和有效性。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，在美国，每年约有20万成人和新生儿患者受到这些综合征的影响，按照目前的护理标准，死亡率高达40%。我们已经开发了一种使用电场的非病毒基因治疗方法，可以在两种小动物模型中预防和治疗现有的ARDS。本研究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更大的受试者，最终转化为人类。在研究结束时，我们将在一个已建立的临床前模型中证明这种新方法治疗肺损伤的有效性。