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

基本信息

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
项目类别：
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

项目摘要

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、破坏性的临床综合征，影响大量成人和新生儿患者。美国每年有 200,000 例病例），按照目前的护理标准，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发生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。该方法简单、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，并开发尽可能简单、安全的方法，以便被临床医生接受。对动物模型的最高推荐是有充分证据证明研究结果在临床试验中相似。为此，我们最近将这种方法应用于一组猪（35-40 公斤），使用的能量低于 0.1 J/ kg，并已实现基因转移到肺部，没有死亡或损伤。我们的 R21 阶段具体目标是 (1) 优化电穿孔参数以实现安全性和将基因转移到猪的肺部，(2) 评估 DNA 递送方法，(3) 开发和优化用于肺基因递送的基于支气管镜的电极，以及(4) 确定 ENaC 以及 Na,K-ATPase 亚基基因的基因转移是否会增加基因疗法治疗小鼠模型肺损伤的功效。在 R21 阶段结束时，我们将建立向猪肺进行基因递送的最佳参数，并确定治疗两个互补的 ALI 猪模型中肺损伤的最佳治疗基因组合。 R33 阶段的具体目标是 (1) 评估电穿孔介导的 Na,K-ATPase/ENaC 基因亚基基因转移在轻中度 ALI 盐水灌洗模型中预防肺损伤的功效，(2) 确定这些转移是否有效基因可以治疗该模型中预先存在的肺损伤，（3）确定这些基因的转移是否可以在严重败血症诱导的 ARDS 盲肠结扎和穿刺模型中保护免受肺损伤，以及(4) 评估该方法是否可用于治疗该模型中先前建立的脓毒症引起的肺损伤。这些研究将为我们提供经胸肺电穿孔的原理验证，并在两个已建立的临床前模型中确定其安全性和有效性。项目叙述急性肺损伤、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破坏性临床综合征，每年影响美国约 200,000 名成人和新生儿患者，按照现行标准，死亡率高达 40%的照顾。我们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。这项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并最终转化为人类。在研究结束时，我们将在已建立的临床前模型中证明这种新方法治疗肺损伤的功效。