Core C: Viral vector core

项目总结（核心C -病毒载体核心） 重组腺相关病毒载体（rAV）介导的HIV抑制剂递送的潜力 在Phil约翰逊，Reed Clark和罗纳德Desrosiers（Nature Medicine 2009），Alex Balazs和大卫巴尔的摩（Nature 2012）， Matthew Gardner和Michael Farzan（Nature 2015）。AAV表达抑制剂的治疗潜力 在该提案和Desrosiers实验室最近发表的工作中清楚地显示了这一点（Immunity 2019）。的 载体核心的主要目的是提供高质量的单链（ss）和自互补（sc） rAAV载体来支持所提出的研究。我们将通过两个具体目标来实现这一目标。目的 1将设计，创建，生产和质量控制测试scAAV载体批次在不同的规模与各种 衣壳，转基因和表达盒，以满足其他研究人员的具体需要，这 程序项目。154只恒河猴将在4年内参加不同的研究， 小鼠将用于前猕猴评价。平均而言，我们估计将有约30个向量批次 每年出版一次，以满足这些研究的需要。Aim将开发一种新型的可扩展的rAAV 用于更大规模的翻译NHP研究和基于rAAV的未来临床开发的生产方法 抗HIV疫苗和治疗剂。我们目前的AAV生产系统应满足本领域的载体需求。 这个项目的前期。然而，大规模的载体生产可能成为一个瓶颈， 更大的转化NHP研究和未来的临床开发。我们将利用我们丰富的经验， 开发各种载体包装细胞系，基于悬浮细胞培养的载体生产， 相应的下游处理和基于色谱的纯化系统，以开发 可扩展的悬浮液293细胞为基础的生产方法，以克服这一限制。为此，我们进一步 我们的共同目标是在猕猴和人类中建立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.