TARGETING THE VIRUS ASSEMBLY AND ENTRY PATHWAYS FOR ANTI-HIV GENE THERAPY

针对抗 HIV 基因治疗的病毒组装和进入途径

• 批准号: 8262811
• 负责人: Anjali Joshi
• 金额: $ 7.55万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-10 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8262811
• 关键词: Adverse effects; Affect; Anti-HIV Agents; CD34 gene; Cell Transplantation; Cells; Complex; Development; Dominant-Negative Mutation; Escape Mutant; Future; Gagging; Gene Transduction Agent; Generations; Genes; Glycoproteins; HIV; HIV Infections; HIV-1; Human; In Vitro; Infection; Integration Host Factors; Interphase Cell; Intervention; Lentivirus Vector; Life Cycle Stages; Mediating; MicroRNAs; Mind; Morphogenesis; Mutate; Mutation; Nature; Pathway interactions; Patients; Pharmacotherapy; Play; Proteins; RNA Binding; RNA Processing; Reporting; Resistance; Role; Salvage Therapy; Sorting - Cell Movement; Staging; Stem cell transplant; Stem cells; TFAP2A gene; Terminally Ill; Testing; Therapeutic Intervention; Toxic effect; Transcription Factor AP-1; Viral; Viral Proteins; Viral Vector; Virion; Virus; Virus Assembly; Virus Replication; cellular transduction; chemotherapy; design; env Gene Products; fitness; gag Gene Products; gene therapy; genetic manipulation; inhibitor/antagonist; mutant; new therapeutic target; novel; particle; rev-Responsive Elements; small hairpin RNA; success; tool

DESCRIPTION (provided by applicant): Targeting the HIV entry and assembly pathways holds promise for development of anti- HIV gene therapy vectors as disrupting the virus at multiple stages in the life cycle will likely limit the emergence of resistant mutants. Moreover, recent advances in human CD34+ stem cell transduction and transplantation have paved a way for anti-HIV gene therapy in the near future. These advances have thus prompted a search for new and potent gene therapy targets for suppression of HIV replication. While si/shRNAs against viral and cellular factors have been tested there are a limited number of studies on dominant negative (DN) viral or cellular proteins that can act as HIV inhibitors. In this regard, the Gag and Envelope proteins of HIV-1 remain an attractive yet unexploited target for anti-HIV gene therapy. Both the Gag and envelope proteins play essential roles in completion of the viral life cycle namely via promoting HIV particle assembly/budding or allowing infection into cells respectively. Notably, virus assembly not only requires the viral Gag protein but also numerous host cell factors like Tsg101, Alix, GGAs, Arfs, POSH, AP-1, AP-3 proteins etc to complete virion morphogenesis. Hence, dominant negative forms of the above genes either individually or in combination have the potential to be developed as powerful tools to target HIV-1 replication. Thus, targeting HIV replication via multiple ways will have the advantage of not only halting virus spread rapidly but also restrict the emergence of resistant isolates. PUBLIC HEALTH RELEVANCE: The rapidly mutating nature of HIV and the adverse effects associated with routine drug therapy solicits the development of alternative therapeutic interventions. One of the obvious alternatives to chemotherapy is gene therapy. Recent advances in genetic manipulation of CD34+ stem cells, along with the development of lentiviral vectors capable of delivering desired genes to non-dividing cells has made HIV-based gene therapy a realistic possibility. An attractive gene therapy approach is to utilize dominant negative (DN) HIV proteins to target HIV life cycle. In this regard the Gag and Envelope (Env) proteins of HIV-1 remain an attractive yet unexploited target because both the Gag and Env proteins are essential components of the virus life cycle and the use of dominant negative forms of these genes individually or in combination have not been thoroughly investigated for anti-HIV gene therapy. In this proposal we aim to identify and design viral vectors expressing DN Gag and Env proteins individually and in combination and test their potential to inhibit WT HIV replication. We also propose to determine whether host factors important for HIV-1 assembly and release like Tsg101 and Alix can be targeted in cells by expression of their DN counterparts and developed as novel gene therapy targets. This approach will not only help identify novel therapeutic targets for HIV intervention but also help decipher the role of known and unknown host factors in the virus replication cycle.

描述（由申请人提供）：针对 HIV 进入和组装途径有望开发抗 HIV 基因治疗载体，因为在生命周期的多个阶段破坏病毒可能会限制耐药突变体的出现。此外，人类CD34+干细胞转导和移植方面的最新进展为不久的将来抗HIV基因治疗铺平了道路。因此，这些进展促使人们寻找新的、有效的基因治疗靶点来抑制艾滋病毒复制。虽然已经测试了针对病毒和细胞因子的 si/shRNA，但针对可充当 HIV 抑制剂的显性失活 (DN) 病毒或细胞蛋白的研究数量有限。在这方面，HIV-1 的 Gag 和 Envelope 蛋白仍然是抗 HIV 基因治疗中一个有吸引力但尚未开发的靶点。 Gag 和包膜蛋白在病毒生命周期的完成中发挥着重要作用，即分别通过促进 HIV 颗粒组装/出芽或允许感染细胞。值得注意的是，病毒组装不仅需要病毒Gag蛋白，​​还需要许多宿主细胞因子，如Tsg101、Alix、GGAs、Arfs、POSH、AP-1、AP-3蛋白等来完成病毒颗粒形态发生。因此，上述基因的显性失活形式无论是单独还是组合都有可能被开发为针对 HIV-1 复制的强大工具。因此，通过多种方式靶向HIV复制不仅具有阻止病毒快速传播的优势，而且还可以限制耐药菌株的出现。 公共卫生相关性：艾滋病毒的快速突变性质以及与常规药物治疗相关的不良反应促使人们开发替代治疗干预措施。化疗的明显替代方法之一是基因治疗。 CD34+干细胞基因操作的最新进展，以及能够将所需基因传递至非分裂细胞的慢病毒载体的开发，使得基于 HIV 的基因治疗成为现实。一种有吸引力的基因治疗方法是利用显性失活 (DN) HIV 蛋白来靶向 HIV 生命周期。在这方面，HIV-1的Gag和包膜(Env)蛋白仍然是一个有吸引力但尚未开发的靶点，因为Gag和Env蛋白都是病毒生命周期的重要组成部分，并且这些基因的显性失活形式单独或组合使用尚未被彻底研究用于抗HIV基因治疗。在本提案中，我们的目标是识别和设计单独和组合表达 DN Gag 和 Env 蛋白的病毒载体，并测试它们抑制 WT HIV 复制的潜力。我们还建议确定对于 HIV-1 组装和释放重要的宿主因子（如 Tsg101 和 Alix）是否可以通过表达其 DN 对应物而靶向细胞中，并开发为新的基因治疗靶点。这种方法不仅有助于确定艾滋病毒干预的新治疗靶点，而且有助于破译已知和未知宿主因素在病毒复制周期中的作用。