Biodegradable Mucus Penetrating DNA Nanoparticle for Gene Therapy of CF

用于 CF 基因治疗的可生物降解粘液穿透 DNA 纳米颗粒

基本信息

批准号：
8863900
负责人：
Justin S. Hanes
金额：
$ 41.21万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8863900
关键词：
Acute Adhesions Adhesives Air Apical Biological Breathing Cells Chronic Clinical Research Clinical Trials Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator DNA DNA delivery Data Dependovirus Diffusion Disease Dose Drug Formulations Effectiveness Endocytosis Epithelial Epithelial Cells Epithelium Esters Future Gastrointestinal tract structure Gel Gene Delivery Gene Expression Gene Mutation Gene Transfer Gene therapy trial Genes Gland Gold Human In Vitro Kinetics Lead Liquid substance Luciferases Lung Lung diseases Macaca Mediating Membrane Modeling Morbidity - disease rate Mucous body substance Mus Non-Viral Vector Osmosis Outcome Penetration Polymers Production Property Proteins Reporter Safety Saline Side Source Sputum Structure of parenchyma of lung Surface Synthetic Genes System Testing Toxic effect Transfection Transgenic Organisms Viral Genes Viral Vector Virus absorption adeno-associated viral vector airway epithelium aqueous base cervicovaginal comparative efficacy cystic fibrosis airway cystic fibrosis mouse cystic fibrosis patients driving force epithelial Na+ channel gene delivery system gene therapy gene therapy clinical trial improved in vivo mortality nanoparticle novel particle prevent public health relevance therapeutic gene transgene expression uptake vector

项目摘要

DESCRIPTION: Cystic fibrosis (CF) gene therapy can potentially benefit all CF patients, regardless of the specific gene mutation leading to the disease state. However, gene therapy trials, using viral and non-viral vectors, have had disappointing outcomes to date due to inability to overcome biological barriers, including the purulent sputum gel layer, periciliary layer and epithelial surface. In particular, we previously found that several clinically and preclinically teted viral and non-viral gene vectors are trapped in human CF sputum, which prevents them from reaching and delivering their therapeutic gene cargo to the underlying airway epithelium. We recently developed two delivery strategies: (i) ultra-small DNA-loaded nanoparticles, called mucus-penetrating DNA nanoparticles (DNA-MPP) that efficiently penetrate human CF sputum and (ii) mildly hypotonic aqueous vehicles, which further improves DNA-MPP penetration through the sputum gel layer and periciliary layer via osmosis-driven fluid absorption, and enhances DNA-MPP uptake by the airway epithelium via the regulatory volume decrease mechanism. We found that the combination of these two approaches led to efficient gene transfer to the airway epithelium of normal mice in vivo and primary human airway cells in vitro. Here we propose to further develop and validate our combined approach in relevant in vitro and in vivo settings. To this end, we will evaluate CFTR gene transfer to CF human primary airway epithelium grown at air-liquid interface and in the lung airways of CF mouse model thoroughly characterized for its CF-like lung diseases. We will also determine whether our delivery strategies allow persistent transgene expression upon a single and repeated dosing without eliciting acute and chronic toxicity. If we are successful, we will test our strategies in macaques and potentially clinical studies in the future.

描述：囊性纤维化（CF）基因疗法可能会受益于所有CF患者，而不论导致疾病状态的特定基因突变。但是，使用病毒和非病毒载体的基因疗法试验由于无能为力而令人失望的结果要克服生物屏障，包括化脓性痰凝胶层，周围层和上皮表面。特别是，我们以前发现，几种临床和先前治疗的病毒和非病毒基因载体被困在人CF痰中，这阻止了它们到达并传递其治疗基因货物到基础气道上皮。我们最近制定了两种输送策略：（i）超小DNA负载的纳米颗粒，称为粘液 - 穿透DNA纳米颗粒（DNA-MPP），可有效地渗透到人类CF痰液中，并（ii）轻度下降水溶液，从通过调节体积减少机制，气道上皮吸收DNA-MPP。我们发现，这两种方法的结合导致体内和原发性人类气道细胞的有效基因转移到正常小鼠的气道上皮。在这里，我们建议在相关的体外和体内环境中进一步开发和验证我们合并的方法。为此，我们将评估CFTR基因转移到在空气界面和CF小鼠模型的肺气道中生长的CF人类原发性气道上皮，这些模型彻底特征了其CF样肺部疾病。我们还将确定我们的输送策略是否允许在单一的反复给药上持续转化表达，而不会引起急性和慢性毒性。如果我们成功，我们将在猕猴中测试我们的策略以及未来的潜在临床研究。