AAV Core

AAV核心

基本信息

批准号：
8311218
负责人：
Guangping Gao
金额：
$ 32.92万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8311218
关键词：
Animals Antibodies Antiviral Agents Autoantigens Biological Assay Blood Circulation Bypass Capsid Cell Culture System Cell Line Cells Clinical Data Development Enrollment Evaluation Future Gene Expression Gene Transfer Goals HIV HIV Antibodies HIV vaccine HIV-1 Human Immunity Immunization In Vitro Infection Intramuscular Injections Macaca Macaca mulatta Massachusetts Mediating Methods Modeling Mus Muscle Neck Population Prevalence Primates Production Proteins Quality Control Recombinant adeno-associated virus (rAAV)Research Personnel Resources Roller Bottle SIV Screening procedure Serological Serotyping Services Staging System Technology Testing Therapeutic Therapeutic Studies Time Transfection Transgenes Tropism Universities Vaccines Viral Viral Vector adeno-associated viral vector base design experience in vivo meetings neutralizing antibody nonhuman primate novel programs simian human immunodeficiency virus therapeutic gene transgene expression vector vector genome

项目摘要

The main objectives of the Vector Core are to provide high quality self-complementary (sc) and single-stranded (ss) rAAV1 vectors and serologically screen NHP animals suitable for rAAV1 anti-viral studies. We will accomplish these goals through the following specific aims: Aim 1. To design, create, produce and quality control test scAAV1 vector lots at different scales with a variety of transgenes and expression cassettes to serve the specific needs of other investigators of this program project. More specifically, 120 rhesus macaques will be enrolled for different studies over 5 years and hundreds of mice will used for pre-macaque evaluation. In average, we estimate that 15-20 vector lots will be produced annually to meet the needs of those studies. Aim 2. To screen NHP populations for pre-existing immunity against AAV1 by using both in vitro and in vivo neutralizing antibody (NAB) assays to select AAV1-NAB free animals for vaccine and therapeutic studies. Pre-screening of NHP population to select the animals without preexisting neutralizing antibody (NAB) to rAAV1 is essential for rAAV1-mediated anti-HIV immunoadhesin gene transfer. Our data suggested that the serological prevalence of primate-derived AAVs in NHP populations ranges from 60-80%. This implies that we may have to screen more than 360 animals to identify 120 animals free of AAV1 NAB. Aim 3. To develop novel and scalable rAAV production method for larger scale translational NHP studies and future clinical development of rAAV1-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 as well. We will utilize our extensive experience in developing various vector packaging cell lines and infection-based vector production system to develop a 293 cell infection-based novel and scalable production method to overcome this limitation.

矢量核心的主要目标是提供高质量的自我平衡（SC）和单链（SS）RAAV1矢量和适合RAAV1抗病毒研究的NHP动物。我们将通过以下特定目标来实现这些目标：目标1。要在不同的尺度上设计，创建，生产和质量控制测试SCAAV1矢量批次，以及各种转基因和表达盒，以满足该计划项目其他研究人员的特定需求。更具体地说，将在5年内进行120个恒河猕猴进行不同的研究，数百只小鼠将用于较长的评估。平均而言，我们估计每年将生产15-20个矢量批次以满足这些研究的需求。目标2。通过使用体外和体内中和抗体（NAB）测定法以选择AAV1-NAB动物进行疫苗和治疗研究，以筛选NHP种群以预先存在免疫力对AAV1。 NHP群体预筛选以选择动物，而无需先前中和抗体（NAB）对RAAV1是RAAV1介导的抗HIV抗HIV免疫粘附基因转移至关重要的。我们的数据表明，NHP种群中灵长类动物衍生的AAV的血清学患病率在60-80％之间。这意味着我们可能必须筛查360多只动物，以识别120只没有AAV1 NAB的动物。目的3。开发新型且可扩展的RAAV生产方法，用于大规模的转化NHP研究以及基于RAAV1的抗HIV疫苗和治疗剂的未来临床发展。我们当前的AAV生产系统应在该计划项目的早期阶段满足向量需求。但是，大规模的矢量产生也可能成为更大的转化NHP研究和未来临床发展的瓶颈。我们将利用我们的丰富经验来开发各种矢量包装细胞系和基于感染的矢量生产系统来开发一种基于293个细胞感染的新颖和可扩展生产方法来克服这种限制。