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的影响很大，但完全控制艾滋病毒-1仍然是一个难以实现的目标。具有抗药性的艾滋病毒-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，HIV-1感染后，2‘，5’-寡腺苷合成酶(2-5OAS)/核糖核酸酶L和p68激酶(PKR)天然抗病毒防御通路受到抑制。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的复制。我们将采用两种策略来实现这一目标：1)PKR和2-5OAS转基因基因的细胞内免疫以保护细胞免受HIV-1感染；2)核酸酶抗性、无毒的2-5A激动剂激活核糖核酸酶L并诱导干扰素和趋化因子的表达。1.以HIV-1为基础的自灭活慢病毒载体(SIN-LV)转导CD34+造血干细胞及其分化的T细胞和单核细胞后，检测抗病毒基因PKR和2-5OAS的表达对HIV-1的抗HIV-1作用。SIN-LV可以转导未分裂细胞和分裂细胞，提高了生物安全性，增强了转基因表达。作为先天抗病毒防御途径的组成部分，PKR和2-5OAS基因产物不受宿主免疫监视或受HIV-1突变的影响。2.检测两种精选的2-5A激动剂对HIV-1血清阳性患者静息状态下的CD4+T细胞的体外抗HIV作用。这些2-5A激动剂在抗病毒防御中通过不同于其他抗HIV-1策略的作用机制绕过HIV-1诱导的阻断。3.通过转导编码PKR和2-5OAS基因的SIN-LV，抑制持续感染的静止CD4+T细胞中重新激活的HIV-1的复制。SIN LV转导细胞与2-5A激动剂或经批准的抗HIV-1药物的联合实验将以相加或协同抗HIV-1活性为目标。这些抗HIV-1策略利用固有的抗病毒防御途径，为抑制HIV-1复制提供了一种新的治疗方法。

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.