AAV-Exosomes: Escaping Neutralizing Antibody and Enhancing Delivery

AAV-外泌体：逃避中和抗体并增强递送

• 批准号: 10428364
• 负责人: Susmita Sahoo
• 金额: $ 79.64万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10428364
• 关键词: Animal Model; Animals; Antibodies; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Cells; Clinic; Clinical; Clinical Data; Clinical Research; Clinical Trials; Data; Defect; Development; Distant; Dose; Encapsulated; Family suidae; Futility; Future; Gene Delivery; Gene Expression; Gene Transduction Agent; Genes; Goals; Heart failure; Human; Immunity; In Vitro; Intravenous Immunoglobulins; Location; Mediating; Mendelian disorder; Methods; Microfluidic Microchips; Modeling; Molecular; Mus; Myocardial; Myocardium; Patients; Pharmacology; Population; Resistance; Rodent; SERCA2a; Safety; Therapeutic; Therapeutic Effect; Tissues; Transduction Gene; Treatment Efficacy; Ultracentrifugation; Viral; Viral Vector; Virus; antibody detection; base; cell type; clinical efficacy; delivery vehicle; exosome; extracellular; gene therapy; gene transfer vector; improved; in vivo; inherited cardiomyopathy; innovation; nanovesicle; neutralizing antibody; pre-clinical; preclinical study; preservation; prevent; success; therapeutic gene; tool; trafficking; transduction efficiency; uptake; vector

SUMMARY Gene therapy is a promising approach for the treatment of various monogenic diseases including inherited cardiomyopathies and other types of heart failure. Adeno-associated vectors (AAV) are vectors of choice for delivering genes to cardiomyocytes for long term expression and due to their safety in clinics. However, a significant challenge to their successful use is futility caused by pre-existing antibodies (NAbs) as well as subsequent development of immunity following AAV administration. NAbs prevent AAVs from infecting target cells, greatly reducing transduction efficiency, and thus, clinical efficacy. Therefore, to advance gene therapies for cardiovascular treatment for a wider population, it is essential to develop strategies to circumvent NAbs. Exosomes are extracellularly secreted nano-vesicles that shuttle selective biomolecules between neighboring and distant cells. Recent studies have shown that exosomes can carry several types viruses and shield them from antibody neutralization. Delivery of AAVs protected by carrier exosomes is a promising approach to circumvent NAb neutralization in AAV-based gene therapy. Our in vitro and in vivo preliminary data suggest that AAV-encapsulating exosomes (AAVExo) are 1) more resistant to NAb neutralization as compared to free AAVs, 2) more efficient in delivering genes to the myocardium, 3) preserves viral cardiotropism, and 4) retains the therapeutic benefits of AAV-mediated gene delivery. Here, we aim to investigate the ability of AAVExo to evade NAbs and serve as a highly efficient gene delivery tool for cardiovascular therapeutics. We have developed a method to isolate highly pure AAVExo with minimum contamination from free-AAVs. Our central hypothesis is that AAVExo shields AAVs to evade NAb and enhance gene delivery to the myocardium compared to free AAVs. Our major goal is to develop a comprehensive understanding of the molecular mechanisms of NAb neutralization by AAVExo, and 2) to determine the beneficial effects of therapeutic genes delivered by AAVExo in preclinical animal models of heart failure. The specific aims are: AIM 1: Characterize AAVExo and determine its gene delivery efficacy and molecular mechanisms of NAb evasion in vitro. AIM 2: Determine the gene delivery efficacy, cardiotropism and mechanisms of NAb evasion of AAVExo in vivo. AIM 3: Optimize AAVExo purification using a NanoDLD microfluidic device and determine the beneficial effect of AAVExo-SERCA2a in small and large animal models of heart failure with preexisting NAb.

概括 基因治疗是一种治疗各种单基因疾病（包括遗传）的有前途的方法 心肌病和其他类型的心力衰竭。腺相关载体（AAV）是首选的向量 将基因传递到心肌细胞以长期表达，并且由于其在诊所的安全性。但是， 对其成功使用的巨大挑战是偶然存在的抗体（NABS）以及 AAV给药后随后发展免疫力。 NABS防止AAV感染 靶细胞，大大降低了转导效率，从而大大降低了临床功效。因此，进步基因 针对更广泛人群的心血管治疗的疗法，制定策略至关重要 绕过nabs。 外泌体是细胞外分泌的纳米紫象，可穿梭选择性生物分子 和遥远的细胞。最近的研究表明，外泌体可以携带几种类型的病毒并屏蔽它们 来自抗体中和。通过载体外泌体保护的AAV是一种有前途的方法 在基于AAV的基因疗法中绕过NAB中和。我们的体外和体内初步数据表明 与游离相比 AAV，2）更有效地将基因输送到心肌，3）保留病毒心脏和4）保留 AAV介导的基因输送的治疗益处。 在这里，我们旨在研究Aavexo逃避NAB并充当高效基因的能力 心血管疗法的递送工具。我们开发了一种隔离高纯avexo的方法 受到自由式AAV的最小污染。我们的中心假设是Aavexo将AAV屏蔽到 与自由AAV相比，逃避NAB并增强了心肌的基因输送。我们的主要目标是 为了全面了解Aavexo的NAB中和的分子机制， 2）确定Aavexo在临床前动物中提供的治疗基因的有益作用 心力衰竭的模型。具体目的是：目标1：表征Aavexo并确定其基因递送 NAB逃避的功效和分子机制在体外。目标2：确定基因递送功效， 心形和逃避体内aavexo的机制。目标3：优化Aavexo纯化 使用Nanodld微流体设备，并确定小型avexo-serca2a的有益效果 大型心力衰竭的大型动物模型与先前存在的NAB。