Next Generation of Recombinant AAV Serotype For Gene Therapy

用于基因治疗的下一代重组 AAV 血清型

基本信息

批准号：
9252509
负责人：
Guangping Gao
金额：
$ 78.86万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-15 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9252509
关键词：
Address Adopted Amino Acids Animal Model Antigen Presentation Antigens Applications Grants Baculoviridae Baculoviruses Biodistribution Biology Blood Circulation Blood Coagulation Disorders CD8B1 gene Canis familiaris Capsid Cell Nucleus Cells Clinical Trials Degradation Pathway Dependovirus Development Disease Dose Engineering Epitopes Excision Factor IX Florida Gene Transfer Hemophilia A Hemophilia B Hepatic Hepatocyte Human I-antigen Immune response Immunobiology Immunology Immunosuppression Investigation Leber&apos s amaurosis Libraries Liver Lymphocyte Lysine Macaca mulatta Methodology Minor Modeling Molecular Evolution Mus Mutate Mutation Patients Performance Peripheral Phosphorylation Portal vein structure Primates Production Program Development Public Health Recombinant adeno-associated virus (rAAV)Research Serine Serotyping Study models Surface System Technology Testing Therapeutic Threonine Time Toxic effect Tropism Tyrosine Ubiquitination Universities Variant Virus adeno-associated viral vector base clinical efficacy combinatorial comparative efficacy cytotoxic design gene therapy humanized mouse immunogenicity improved in vivo interdisciplinary approach mouse model multicatalytic endopeptidase complex neutralizing antibody next generation nonhuman primate novel public health relevance response scale up stable cell line therapeutic gene tissue tropism trafficking transduction efficiency vector virology

项目摘要

DESCRIPTION: The main aims of this multiple-PI proposal are: to continue to define the underlying mechanisms of improved hepatocyte transduction by our next generation of recombinant adeno-associated virus (AAV) vectors; to extend these studies to novel AAV2 variants with high liver tropism, enriched from a library, and to AAV3-based vectors, which show high efficacy in human hepatocytes and non-human primate liver; and to evaluate the efficacy and host immune response in gene therapy for hemophilia B. Candidate vectors for clinical trial with high efficacy in small and large animal models will be identified, with particular emphasis on in vivo gene transfer to human/primate hepatocytes and immunobiology. The current application encompassing an interdisciplinary approach should allow us to develop superior next generation vectors for liver-directed human gene therapy, drawing heavily from expertise at the University of Florida in AAV vector biology, immunology of AAV gene transfer, and animal model studies. We will test the following hypotheses: a. Minor changes in the capsid sequence can substantially improve liver gene transfer, in part through postentry mechanisms, allowing us to design superior AAV2- and AAV3-based vectors. b. A combination of molecular evolution and capsid engineering will result in vectors with high transduction efficiency of human/primate hepatocytes and reduced MHC I antigen presentation. c. Capsid-modified AAV vectors will provide therapeutic F.IX expression at low vector doses and with reduced immunogenicity. The following three Specific Aims will be pursued: Specific Aim 1: Further optimize and compare in vivo performance of novel AAV2- and AAV3-based vectors, and define the mechanisms responsible for the robust liver gene transfer. Specific Aim 2: Identify novel capsid variants for high-efficiency in vivo gene transfer to human hepatocytes and adopt these into an advanced baculovirus production system. Specific Aim 3: Test improved vectors in animal models of hemophilia and in non-human primates, and define their immunogenicity. These investigations have a high chance of yielding superior vectors for liver gene transfer in humans.

描述：多个PI的主要目的是定义我们的T生成的重组腺相关病毒（AAV）载体的肝细胞转导的基本机制，以将研究扩展到具有高肝脏症的新型AAV2变体中图书馆和基于AAV3的载体，人类肝细胞和非人类灵长类动物肝脏中的WHMPITOS；确定，特别强调体内基因转移到人/灵长类动物肝细胞和免疫生物学。肝脏基因转移，部分通过邮政机制，使我们能够设计基于AAV2的载体。在低载体矢量剂量上提供治疗性f.ix表达，并以降低的免疫原性来提供以下三个特定目标：特定目标1：进一步优化和基于AAV3的矢量，并定义了负责鲁棒的机制肝脏基因转移：识别用于人体基因的新型衣壳变异，并在系统上采用晚期杆菌，并在非人类动物的动物模型中测试改进的载体。这些研究很有可能在人类中进行肝基因转移的产量向量。