Core C: Viral vector core

核心C：病毒载体核心

基本信息

批准号：
10381476
负责人：
Guangping Gao
金额：
$ 24.15万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-15 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10381476
关键词：
Animals Antibodies Autoantigens Baltimore Blood Circulation Bypass Capsid Cell Culture System Cell Culture Techniques Cell Line Cells Chromatography DNA cassette Data Enrollment Evaluation Future Gene Expression Gene Transfer Goals HIV HIV Infections HIV vaccine Human Immunity Immunization Infection Intramuscular Injections Macaca Macaca mulatta Mediating Medicine Methods Modeling Mus Muscle Nature Neck Production Proteins Publishing Quality Control Recombinant adeno-associated virus (rAAV)Research Personnel Rhesus Roller Bottle SIV Serotyping Suspensions System Technology Testing Therapeutic Time Transfection Transgenes Tropism Vaccine Design Viral Vector Work adeno-associated viral vector base clinical development design experience in vivo inhibitor nonhuman primate novel programs transgene expression vaccine trial vector vector genome

项目摘要

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.

项目摘要（核心 C – 病毒载体核心）重组腺相关病毒载体 (rAAV) 介导的 HIV 抑制剂递送的潜力 Phil Johnson、Reed 的工作清楚地证明了对 HIV 感染的广泛中和活动 Clark 和 Ronald Desrosiers（《自然医学》2009 年）、Alex Balazs 和 David Baltimore（《自然》2012 年）、 Matthew Gardner 和 Michael Farzan（Nature 2015）。AAV 表达抑制剂的治疗潜力。该提案和 Desrosiers 实验室最近发表的工作（Immunity 2019）清楚地表明了这一点。 Vector Core 的主要目标是提供高质量的单链 (ss) 和自互补 (sc) rAAV 载体支持拟议的研究，我们将通过两个具体目标来实现这一目标。 1 将设计、创建、生产和质量控制测试不同规模的 scAAV 载体批次，包括各种衣壳、转基因和表达盒，以满足该领域其他研究人员的特定需求计划项目将招募 154 只恒河猴进行为期 4 年的不同研究，其中包括数百只恒河猴。小鼠将用于猕猴前评估平均而言，我们估计将有 -30 个向量批次。每年生产以满足这些研究的需求，Aim 将开发一种新颖且可扩展的 rAAV。用于更大规模转化 NHP 研究和基于 rAAV 的未来临床开发的生产方法我们目前的 AAV 生产系统应该能够满足载体的需求。然而，大规模载体生产可能成为该计划项目的瓶颈。我们将利用我们在更大的转化 NHP 研究和未来的临床开发方面的丰富经验。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，以开发基于可扩展悬浮293细胞的生产方法克服了这一限制。我们的共同目标是在猕猴和人类中建立 AAV 介导的功能性治疗方法。