Episomal lenti vector for a humanized hemophilia mouse

人源化血友病小鼠的游离慢病毒载体

• 批准号: 7663777
• 负责人: TAL KAFRI
• 金额: $ 26.29万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7663777
• 关键词: Address; Biodistribution; Biological Assay; Cells; Chimera organism; Chromatin Structure; Complementary DNA; Development; Equine Infectious Anemia Virus; Exhibits; Factor IX; Gene Delivery; Genetic; Genome; Goals; HIV-1; HIV-1 integrase; Hemophilia A; Hepatic; Hereditary Disease; Human; Image; Immune response; Insertional Mutagenesis; Interphase Cell; Laboratories; LacZ Genes; Lentivirus Vector; Liver; Luciferases; Mediating; Messenger RNA; Modality; Molecular; Mus; Nature; Nuclear Export; Oncogenic; Patients; Process; Production; Protocols documentation; RNA Binding; Reporter; Research Proposals; Risk; Signal Transduction; Specificity; System; Testing; Therapeutic; Time; Viral Vector; base; cellular transduction; chromatin modification; efficacy testing; env Gene Products; gene replacement; gene replacement therapy; gene therapy; histone modification; improved; in vivo; inhibitor/antagonist; microbiological attachment sites; mouse model; mutant; novel; particle; prevent; transduction efficiency; transgene expression; vector; vector-induced

DESCRIPTION (provided by applicant): The spectrum of genetic diseases that can be therapeutically addressed by using lentiviral vectors is restricted by their oncogenic potential, which is inherent to their integrative nature. In this study we propose to develop and utilize novel non-integrating lentiviral vectors to address the above obstacle. Our approach is based on recent findings in our laboratory demonstrating the ability of the HIV-1 Rev/RRE system to function as a secondary packaging system, which mediates packaging of non-HIV-1 mRNAs into HIV-1 particles. We showed that HIV-1 RRE containing EIAV vectors efficiently packaged into HIV-1 particles. The novel chimeric vectors, which lacked the ability to integrate into a host cells' genome, exhibited transgene expression levels significantly higher than other non-integrating vectors, such as HIV-1 vectors packaged with the HIV-1 integrase mutant E152A. Here we propose to: i) characterize the mechanism involved in the Rev/RRE dependent packaging of chimeric EIAV/HIV-1 vectors; ii) characterize the ability of the HIV-1 to process the EIAV att sites and its effects on chimera vector integration; iii) investigate the effects of histone modifications and chromatin structure on transgene expression from EIAV/HIV-1 chimera vectors; iv) to characterize the biodistribution of the chimera EIAV/HIV-1 vectors; and v) to determine the efficacy of the chimera vectors at delivering and maintaining high levels of human factor IX expression in a humanized hemophilia mouse model We believe that the proposed studies will result in the development of efficacious non-integrating lentiviral vectors, which will be better suited for human gene therapy.

描述（由申请人提供）：可以通过使用慢病毒载体治疗解决的遗传疾病的范围受到其致癌潜力的限制，这是其综合性质所固有的。在本研究中，我们建议开发和利用新型非整合慢病毒载体来解决上述障碍。我们的方法基于我们实验室的最新发现，该发现证明了 HIV-1 Rev/RRE 系统具有作为二级包装系统的功能，该系统介导非 HIV-1 mRNA 包装成 HIV-1 颗粒。我们表明，含有 EIAV 载体的 HIV-1 RRE 可以有效地包装成 HIV-1 颗粒。这种新型嵌合载体缺乏整合到宿主细胞基因组中的能力，其转基因表达水平显着高于其他非整合载体，例如包装有HIV-1整合酶突变体E152A的HIV-1载体。在这里，我们建议： i) 描述嵌合 EIAV/HIV-1 载体 Rev/RRE 依赖性包装所涉及的机制； ii) 表征 HIV-1 处理 EIAV att 位点的能力及其对嵌合载体整合的影响； iii) 研究组蛋白修饰和染色质结构对 EIAV/HIV-1 嵌合载体转基因表达的影响； iv) 表征嵌合体 EIAV/HIV-1 载体的生物分布； v) 确定嵌合载体在人源化血友病小鼠模型中递送和维持高水平的人类因子IX表达的功效我们相信，所提出的研究将导致有效的非整合慢病毒载体的开发，这将更适合人类基因治疗。