Evolving High Potency AAV Vectors for Neuromuscular Genome Editing

进化用于神经肌肉基因组编辑的高效 AAV 载体

• 批准号: 10465740
• 负责人: Aravind Asokan
• 金额: $ 83.36万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-10 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10465740
• 关键词: Evolving; Potency; AAV; Vectors; Neuromuscular

ABSTRACT Recombinant adeno-associated viruses (AAV) have emerged as safe and effective vectors for clinical gene therapy applications including systemic treatment of neuromuscular diseases such as Spinal Muscular Atrophy (SMA), Duchenne Muscular Dystrophy (DMD), and Giant Axonal Neuropathy (GAN) amongst others. However, enabling gene editing therapeutics to treat human disease requires improved systems that achieve effective genome editing at low systemic AAV vector doses in multiple animal models. Additionally, the ideal target cell types for gene editing, including progenitor cells, are distinct from the target cells that conventional gene delivery vectors have been optimized for. Genome editing in neuromuscular tissue, in particular, is challenging. To date, no effective non-viral delivery vehicles have been identified for widespread genome editing in musculoskeletal and neurological tissue following systemic administration. Further, in case of AAV vectors, several challenges pertinent to delivery of genome editors remain. The rationale for our current proposal stems from barriers posed by (i) species-related differences observed in AAV tropism, (ii) presence of pre-existing neutralizing antibodies to natural AAV, (iii) the need for high systemic AAV doses to achieve neuromuscular gene transfer, (iv) vector dose-related toxicity as indicated by detection of liver transaminases in patients, and (v) the unique opportunity to correct progenitor cells in situ. To address these aspects, we have assembled a collaborative team with cross- cutting expertise and developed a comprehensive and innovative approach to evolve high potency AAV variants for systemic neuromuscular genome editing. Specifically, we will focus our efforts on evolving high potency AAV vectors for neuromuscular genome editing using a three-tiered approach applied across different species, tissues and cell types.

抽象的 重组腺相关病毒（AAV）已成为安全有效的临床基因载体 治疗应用，包括神经肌肉疾病（如脊髓性肌肉萎缩症）的全身治疗 (SMA)、杜氏肌营养不良症 (DMD) 和巨轴突神经病 (GAN) 等。然而， 使基因编辑疗法能够治疗人类疾病需要改进系统以实现有效 在多种动物模型中以低全身 AAV 载体剂量进行基因组编辑。此外，理想的靶细胞 用于基因编辑的类型，包括祖细胞，与传统基因传递的靶细胞不同 载体已经过优化。神经肌肉组织的基因组编辑尤其具有挑战性。迄今为止， 尚未发现有效的非病毒递送载体可用于肌肉骨骼中广泛的基因组编辑 和全身给药后的神经组织。此外，就 AAV 载体而言，还存在一些挑战 与基因组编辑器的交付相关的问题仍然存在。我们当前提案的理由源于所构成的障碍 通过 (i) AAV 趋向性观察到的物种相关差异，(ii) 预先存在的中和抗体的存在 天然 AAV，(iii) 需要高全身 AAV 剂量才能实现神经肌肉基因转移，(iv) 载体 通过检测患者的肝脏转氨酶表明剂量相关的毒性，以及（v）独特的机会 原位纠正祖细胞。为了解决这些问题，我们组建了一个跨部门协作团队 切割专业知识并开发了一种全面的创新方法来进化高效 AAV 变体 用于系统性神经肌肉基因组编辑。具体来说，我们将集中精力开发高效 AAV 使用适用于不同物种、组织的三层方法进行神经肌肉基因组编辑的载体 和细胞类型。