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：
9229059
负责人：
Justin S. Hanes
金额：
$ 39.64万
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/9229059
关键词：
Acute Adhesions Adhesives Air Apical Biological Breathing Cells Chronic Clinical Clinical Research Clinical Trials Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator DNA DNA delivery Data Dependovirus Diffusion Disease Dose Effectiveness Endocytosis Epithelial Epithelial Cells Epithelium Esters Formulation Future Gastrointestinal tract structure Gel Gene Delivery Gene Expression Gene Mutation Gene Transfer Gene therapy trial Genes Gland Gold Human Hydration status 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 Virus absorption acute toxicity adeno-associated viral vector airway epithelium aqueous base cervicovaginal comparative efficacy cystic fibrosis airway cystic fibrosis patients epithelial Na+ channel gene delivery system gene therapy gene therapy clinical trial improved in vivo mortality mouse model nanoparticle novel overexpression particle pre-clinical 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) 基因治疗可能使所有 CF 患者受益，无论导致疾病状态的特定基因突变如何。然而，迄今为止，使用病毒和非病毒载体的基因治疗试验的结果令人失望。无力克服生物屏障，包括化脓性痰凝胶层、纤毛周围层和上皮表面。特别是，我们之前发现一些临床和临床前测试的病毒和非病毒基因载体被困在人类CF痰中，这阻止了它们到达和传播。我们最近开发了两种递送策略：(i) 超小型 DNA 负载纳米颗粒，称为“纳米粒子”。粘液穿透性 DNA 纳米颗粒 (DNA-MPP) 可有效穿透人类 CF 痰液，并且 (ii) 轻度低渗水性载体可通过渗透驱动的液体吸收进一步提高 DNA-MPP 对痰液凝胶层和纤毛周围层的渗透性，并增强 DNA -气道上皮通过调节体积减少机制摄取MPP 我们发现这两种方法的结合导致了基因有效转移到气道。在此，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的组合方法，为此，我们将评估 CFTR 基因转移到 CF 人初级气道上皮。我们还将确定我们的递送策略是否允许在单次和重复给药后持续转基因表达，而不引起急性和慢性。如果我们成功了，我们将在猕猴身上测试我们的策略。以及未来潜在的临床研究。