Development of AAV vectors for CF therapy

用于 CF 治疗的 AAV 载体的开发

• 批准号: 10117463
• 负责人: Chengwen Li
• 金额: $ 38.88万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10117463
• 关键词: Address; Airway Disease; Animal Model; Binding; Blindness; Blood; Blood Coagulation Disorders; Blood Vessels; CRISPR/Cas technology; Capsid; Clinical Trials; Complication; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; DNA; Dependovirus; Development; Directed Molecular Evolution; Disease; Epithelial Cells; Failure; Gene Delivery; Genes; Genetic Diseases; Genetic Engineering; Genome; Hemorrhage; High Prevalence; Human; In Vitro; Incubated; Intravenous Immunoglobulins; Libraries; Longitudinal Studies; Lung; Lung diseases; Measurable; Mediating; Morbidity - disease rate; Mus; Mutation; Organ; Patients; Peptides; Peripheral; Phage Display; Phenotype; Plant Roots; Property; Proteins; Regulator Genes; Serotyping; Technology; Testing; Tissues; Transduction Gene; Variant; Vascular Permeabilities; Vectorial capacity; Viral Genome; Viral Packaging; Virion; adeno-associated viral vector; airway epithelium; apical membrane; base; basolateral membrane; cellular transduction; cystic fibrosis airway epithelia; cystic fibrosis mouse; cystic fibrosis patients; ds-DNA; early phase clinical trial; gene therapy; mortality; mutant; neutralizing antibody; novel; novel strategies; pulmonary function; recessive genetic trait; small molecule; success; transduction efficiency; vector; vector-induced

Abstract Cystic fibrosis (CF) is caused by mutations of the CF transmembrane conductance regulator (CFTR) gene. Airway disease is the leading cause of morbidity and mortality. Gene therapy offers a potential cure for CF. Among gene delivery vehicles, adeno-associated virus (AAV) vectors have demonstrated success in clinical trials. AAV vectors have also been tested in CF patients via airway delivery for gene addition but without measurable success perhaps due to physical airway barriers and inefficient transduction. Studies have shown that higher AAV transduction of airway epithelial cells occur via the basolateral membrane in vitro and our preliminary study demonstrated that systemic administration of AAV vectors induced much more efficient lung transduction than airway delivery in mice. These findings strongly suggest that AAV can transduce airway epithelial cells via systemic administration. The efficiency of AAV transduction in airway epithelial cells after systemic delivery of AAV vectors is usually restricted by the blood vascular barrier and a high prevalence of AAV neutralizing antibodies (Nabs). In this proposal, based on our previous studies, we will explore different strategies to develop novel AAV vectors using directed evolution and AAV virion specific binding peptides via phage display technology to increase AAV vector vascular permeability for enhanced transduction in airway epithelial cells and to evade Nabs after systemic administration. Finally, the novel approaches developed in this proposal will be used to deliver optimized CRISPR/Cas9 specific for CFTR del508 to CF mice and study the long-term phenotypic correction. The results generated from these studies will allow us to develop an effective approach to treat CF patients using AAV vector mediated gene delivery. .

抽象的 囊性纤维化 (CF) 是由 CF 跨膜电导调节因子 (CFTR) 基因突变引起的。 气道疾病是发病和死亡的主要原因。基因疗法为 CF 提供了潜在的治愈方法。 在基因递送载体中，腺相关病毒（AAV）载体已在临床上取得了成功 试验。 AAV 载体还通过气道递送在 CF 患者中进行了基因添加测试，但没有 可衡量的成功可能是由于物理气道障碍和低效的转导。研究表明 在体外，气道上皮细胞的 AAV 较高转导是通过基底外侧膜发生的，我们的 初步研究表明，全身施用 AAV 载体可诱导更有效的肺功能。 小鼠中的转导优于气道递送。这些发现强烈表明 AAV 可以转导气道 上皮细胞通过全身给药。气道上皮细胞中 AAV 转导效率 AAV 载体的全身递送通常受到血管屏障和高患病率的限制 AAV 中和抗体 (Nab)。在本提案中，基于我们之前的研究，我们将探索不同的 使用定向进化和 AAV 病毒体特异性结合肽开发新型 AAV 载体的策略 噬菌体展示技术可增加 AAV 载体血管通透性，从而增强气道转导 上皮细胞并在全身给药后逃避 Nabs。最后，开发的新颖方法 该提案将用于向 CF 小鼠传递针对 CFTR del508 的优化 CRISPR/Cas9 并进行研究 长期表型校正。这些研究产生的结果将使我们能够开发一种 使用 AAV 载体介导的基因传递治疗 CF 患者的有效方法。 。