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

描述（由申请人提供）：尽管抗逆转录病毒药物具有重大影响，但完全控制HIV-1仍然是一个难以实现的目标。抗药性艾滋病毒-1变种的出现和潜伏的艾滋病毒-1宿主的重新激活仍然是巨大的障碍。此外，2 '，5'-寡腺苷酸合成酶（2- 5 OAS）/RNA酶L和p68激酶（PKR）先天抗病毒防御途径在HIV-1感染后被抑制。该项目的工作假设是，恢复这些抗病毒防御途径可以提供一种有效的治疗方法来抑制HIV-1复制。我们将应用两种策略来实现这一目标：1）PKR和2- 5 OAS转基因的细胞内免疫以保护细胞免受HIV-1感染，以及2）核酸酶抗性、无毒的2-5A激动剂，其激活RNA酶L并诱导干扰素和趋化因子表达。具体目标是：1.确定抗病毒转基因PKR和2- 5 OAS在转导的CD 34+造血干细胞及其分化的T细胞和单核细胞后代中表达的抗HIV-1效应，这些细胞在通过基于HIV-1的自失活慢病毒载体（SIN LV）递送后发生。SIN LV可以使非分裂和分裂细胞具有增加的生物安全性和增强的转基因表达。作为先天性抗病毒防御途径的组成部分，PKR和2- 5 OAS基因产物不受宿主免疫监视或受HIV-1突变的影响。2.确定两种选择的2-5A激动剂在来自病毒血症和病毒血症HIV-1血清阳性患者的静息CD 4 + T淋巴细胞中的体外抗HIV作用。这些2-5A激动剂通过不同于其他抗HIV-1策略的作用机制来规避HIV-1诱导的抗病毒防御阻断。3.通过转导编码PKR和2- 5 OAS转基因的SIN LV抑制持续感染的静息CD 4 + T细胞中再活化的HIV-1的复制。将进行SIN LV转导的细胞与2-5A激动剂或批准的抗HIV-1药物的组合实验，目的是增加或协同抗HIV-1活性。这些利用先天性抗病毒防御途径的抗HIV-1策略为抑制HIV-1复制提供了新的治疗方法。

DESCRIPTION (provided by applicant): 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-1 from 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.