Core C: Viral vector core

项目摘要(核心C-病毒载体核心) 重组腺相关病毒载体(RAAV)介导人免疫缺陷病毒(HIV)抑制物基因治疗的研究 里德·菲尔·约翰逊的研究清楚地证明了广泛的中和艾滋病毒感染的活性 Clark和Ronald Desrosiers(《自然医学》2009)，Alex Balazs和David Baltimore(《自然》2012)， 马修·加德纳和迈克尔·法赞(《自然》2015)。AAV表达的抑制物的治疗潜力 这一建议和Desrosiers实验室最近发表的工作(豁免2019年)清楚地表明了这一点。这个 载体核心的主要目标是提供高质量的单链(Ss)和自我互补(Sc) RAAV载体以支持拟议的研究。我们将通过两个具体目标实现这一目标。目标 1将设计、创建、生产和质量控制不同规模的scAAV载体批次，并使用各种 衣壳、转基因和表达盒，以满足其他研究人员的特定需要 计划项目。154只恒河猴将在4年内参加不同的研究，数百只 老鼠将被用来进行猕猴前评估。平均而言，我们估计-30个向量地块将是 每年制作，以满足这些研究的需要。AIM将开发一种新颖且可扩展的rAAV 基于rAAV的大规模翻译NHP研究的生产方法和未来的临床发展 抗艾滋病毒疫苗和治疗药物。我们目前的AAV生产系统应该能满足 该计划项目的早期阶段。然而，大规模的媒介生物生产可能会成为阻碍 更大的翻译NHP研究和未来的临床开发。我们将利用我们丰富的经验在 开发各种载体包装细胞系，以悬浮细胞培养为基础的载体生产， 相应的下游处理和基于层析的纯化系统，以开发 可伸缩悬浮式293电池生产方法克服了这一限制。通过这样做，我们进一步 我们的共同目标是在猕猴和人类身上建立AAV介导的功能性治疗。

PROJECT SUMMARY (Core C – Viral Vector Core) The potential of recombinant adeno-associated virus vector (rAAV)-mediated delivery of HIV inhibitors with broadly neutralizing activities to HIV infection is clearly demonstrated in work from Phil Johnson, Reed Clark, and Ronald Desrosiers (Nature Medicine 2009), Alex Balazs and David Baltimore (Nature 2012), Matthew Gardner and Michael Farzan (Nature 2015). The therapeutic potential of AAV-expressed inhibitors is clearly shown in this proposal and recent published work from the Desrosiers Lab (Immunity 2019). The main objective of the Vector Core is to provide high quality single-stranded (ss) and self-complementary (sc) rAAV vectors to support the proposed studies. We will accomplish this goal through two specific aims. Aim 1 will design, create, produce, and quality control test scAAV vector lots at different scales with a variety of capsids, transgenes and expression cassettes to serve the specific needs of other investigators of this program project. 154 rhesus macaques will be enrolled for different studies over 4 years, and hundreds of mice will be used for pre-macaque evaluation. On average, we estimate that -30 vector lots will be produced annually to meet the needs of those studies. Aim will develop a novel and scalable rAAV production method for larger scale translational NHP studies and future clinical development of rAAV-based anti-HIV vaccine and therapeutics. Our current AAV production system should meet the vector needs in the early stage of this program project. However, large scale vector production may become a bottle neck for larger translational NHP studies and future clinical development. We will utilize our extensive experience in developing various vector packaging cell lines, suspension cell culture-based vector production, corresponding downstream processing and chromatography-based purification systems to develop a scalable suspension 293 cell-based production method to overcome this limitation. In doing so, we further our common goal of establishing AAV-mediated functional cures in macaques and humans.