HIV-1 Inhibition by Delivery of 2-5OAS & PKR Genes

尽管抗逆转录病毒药物产生了重大影响，但完全控制HIV-1仍然是一个难以实现的目标。 抗药性HIV-1变异体的出现和潜伏HIV-1宿主的重新激活仍然令人生畏 障碍.此外，2 '，5'-寡腺苷酸合成酶（2 - 5OAS）/RNase L和p68激酶（PKR）的先天性 抗病毒防御途径在HIV-1感染后被抑制。这个项目的工作假设是 这些抗病毒防御途径的恢复可以提供一种有效的治疗方法， HIV-1复制。我们将采用两种策略来实现这一目标：1）PKR的细胞内免疫， 2-5 OAS转基因保护细胞免受HIV-1感染和2）核酸酶抗性，无毒2 - 5A激动剂 其激活RNase L并诱导干扰素和趋化因子表达。具体目标是： 1.为了确定抗病毒转基因PKR和2 - 5OAS在小鼠中表达的抗HIV-1作用， 转导的CD34+造血干细胞及其分化的T细胞和单核细胞后代 随后通过基于HIV-1的自失活慢病毒载体（SIN LV）递送。SIN LVs可以 具有增加的生物安全性和增强的转基因表达的非分裂和分裂细胞。作为组件 在天然抗病毒防御途径中，PKR和2 - 5OAS基因产物不受宿主免疫应答的影响， 监测或受HIV-1突变的影响。 2.确定两种选择的2 - 5A激动剂在静息CD4 + T淋巴细胞中的体外抗HIV作用 来自病毒血症和病毒血症HIV-1血清阳性患者。这些2 - 5A激动剂规避HIV-1诱导的 通过不同于其他抗HIV-1策略的作用机制阻断抗病毒防御。 3.抑制HIV-1从持续感染的静息CD4 + T细胞中的复制， 用编码PKR和2 - 5OAS转基因的SIN LV转导。SIN LV-1的组合实验 用2 - 5A激动剂或批准的抗HIV-1药物转导的细胞将以相加或 协同抗HIV-1活性。这些抗HIV-1的策略，利用先天的抗病毒防御， 途径，提供了一种新的治疗方法来抑制HIV-1的复制。

Complete control of HIV-1 remains an elusive goal despite the significant impact of antiretroviral drugs. Emergence of drug-resistant HIV-1 variants and the reactivation of latent HIV-1 reservoirs remain formidable obstacles. Furthermore, the 2',5'-oligoadenylate synthetase (2-5OAS)/RNase L and p68 kinase (PKR) innate antiviral defense pathways are inhibited following HIV-1 infection. The working hypothesis of this project is that restoration of these antiviral defense pathways can provide an effective therapeutic approach to inhibit HIV-1 replication. We will apply two strategies to achieve this goal: 1) intracellular immunization of PKR and 2-5OAS transgenes to protect cells against HIV-1 infection and 2) nuclease-resistant, non-toxic 2-5A agonists that activate RNase L and induce interferon and chemokine expression. The specific aims are: 1. To determine the anti-HIV-1 effects of expression of the antiviral transgenes, PKR and 2-5OAS, in transduced CD34+ hematopoietic stem cells and their differentiated T cell and monocyte progeny following delivery by HIV-1 based self-inactivating lentiviral vectors (SIN LVs). SIN LVs can transduce non-dividing and dividing cells with increased biosafety and enhanced transgene expression. As components of the innate antiviral defense pathways, the PKR and 2-5OAS gene products are not subject to host immune surveillance or affected by mutations in HIV-1. 2. To determine the in vitro anti-HIV effects of two select 2-5A agonists in resting CD4+ T lymphocytes from viremic and aviremic HIV-1 seropositive patients. These 2-5A agonists circumvent HIV-1 induced blockades in antiviral defense by mechanisms of action distinct from other anti HIV-1 strategies. 3. To inhibit replication of reactivated HIV-1from persistently infected resting CD4+ T cells by transduction with SIN LVs encoding PKR and 2-5OAS transgenes. Combination experiments of SIN LV- transduced cells with 2-5A agonists or approved anti-HIV-1 drugs will be conducted with the goal of additive or synergistic anti-HIV-1 activity. These anti-HIV-1 strategies, which utilize the innate antiviral defense pathways, offer a new therapeutic approach to the inhibition of HIV-1 replication.