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

描述(申请人提供)：以艾滋病毒进入和组装途径为目标有望开发抗艾滋病毒基因治疗载体，因为在生命周期的多个阶段干扰病毒可能会限制耐药突变的出现。此外，人类CD34+干细胞转导和移植的最新进展为在不久的将来进行抗HIV基因治疗铺平了道路。因此，这些进展促使人们寻找新的、有效的基因治疗目标，以抑制艾滋病毒的复制。虽然针对病毒和细胞因子的si/shRNAs已经被测试，但关于显性阴性(DN)病毒或细胞蛋白可以作为HIV抑制物的研究数量有限。在这方面，HIV-1的Gag和包膜蛋白仍然是抗HIV基因治疗的一个有吸引力但尚未开发的靶点。Gag蛋白和包膜蛋白在病毒生命周期的完成中都发挥着重要作用，即分别通过促进HIV颗粒组装/萌发或允许感染进入细胞。值得注意的是，病毒的组装不仅需要病毒的Gag蛋白，还需要大量的宿主细胞因子，如Tsg101、Alix、ggas、ARFS、POSH、AP-1、AP-3蛋白等来完成病毒粒子的形态发生。因此，上述基因的显性阴性形式，无论是单独的还是组合的，都有可能被开发为针对HIV-1复制的强大工具。因此，通过多种方式针对艾滋病毒复制将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。

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.