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

描述（由申请人提供）：靶向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HIV复制。虽然已经测试了针对病毒和细胞因子的si/ shrna，但对可作为HIV抑制剂的显性阴性（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基因治疗的一个有吸引力但尚未开发的靶标。Gag蛋白和包膜蛋白都在病毒生命周期的完成过程中发挥着重要作用，即分别通过促进HIV颗粒组装/出芽或允许感染进入细胞。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，都有可能成为靶向HIV-1复制的强大工具。因此，通过多种途径靶向HIV复制不仅具有迅速阻止病毒传播的优势，而且还具有限制耐药分离株出现的优势。

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.