Blocking HIV with aptamers targeted to viral components

描述(由申请人提供)：对艾滋病毒复制的不完全抑制和随之而来的耐药性的发展继续损害艾滋病治疗。潜在的问题包括疗效、不粘连和受感染的细胞库。尽管针对进入、整合或转录的新药将改善一些问题，但要想获得更完整的解决方案，就必须转向新的复合疗法。在针对HIV-1的几种有效的基因治疗方法中，针对HIV-1的RNA适配子是可用的。适配子以其特异性、高亲和力、稳定性和缺乏免疫原性而闻名。最近的工作提供了令人信服的证据，即针对HIV-1的细胞内适配子可以强烈抑制病毒复制。对这种适配子产生抗药性的突变导致了病毒适应性的丧失。因此，有一个迄今尚未利用的机会来开发具有独特特异性(降低毒性)的抗艾滋病毒药物，这种药物将更完全地抑制艾滋病毒-1复制，阻碍或减缓耐药性，并消除不粘连。为此，提出了涉及各自组织/调查人员的以下合作研究方案。1.Accacia LLC，Int(德克萨斯州奥斯汀)in.与安迪·埃灵顿(德克萨斯大学奥斯汀分校)合作，将高通量地选择针对HIV-1靶标的RNA适配子，识别紧密结合，确定并进一步优化体外效果。2.Vinayaka Prasad(AECOM)将使用来自Accacia的预选适配子来确定抑制HIV/SIV复制的有效性，确定抑制机制，并选择适体抗性。他将为Accacia提供抗药性蛋白质，用于开发第二代适配子。3.Paul Johnson(NEPRC)将通过逆转录病毒载体将最好的适配子引入猕猴CD34+Ve细胞，将其移植到猕猴体内，检测未感染猕猴的基因标记水平，并测试该适配子在体内保护来自标记的CD34+Ve细胞的CD4+ve T细胞的效果。

DESCRIPTION (provided by applicant): Incomplete suppression of HIV replication and consequent development of resistance continue to mar AIDS therapy. The underlying problems include efficacy, non-adherence and infected cell reservoirs. Although new drugs targeting entry, integration or transcription will ameliorate some problems, achieving a more complete solution necessitates turning to novel and compementary therapies. Among several, effective gene therapy approaches available for HIV-1 are RNA aptamers targeted to HIV-1. Aptamers are known for their specificity, high affinity, stability and the absence of immunogenicity. Recent work has provided convincing evidence that intracellular aptamers targeted to HIV-1 can strongly suppress viral replication. Mutations conferring resistance to such aptamers have led to loss of viral fitness. Thus, there is a hitherto un-utilized opportunity to develop anti-HIV agents of unique specificity (reducing toxicity), that would more completely suppress HIV-1 replication, hinder or slow-down resistance and eliminate non-adherence. To this end, the following collaborative research program involving the respective organizations/investigators is proposed. 1. Accacia LLC, Int (Austin, TX) in. partnership with Andy Ellington (University of Texas, Austin), will perform high throughput selection of RNA aptamers to HIV-1 targets, identify tight-binders, determine and further optimize in vitro efficacies. 2. Vinayaka Prasad (AECOM) will use pre-selected aptamers from Accacia to determine efficacy of inhibition of HIV/SHIV replication, determine mechanism of inhibition and select for aptamer-resistance. He will provide Accacia with resistant proteins for developing second generation aptamers. 3. Paul Johnson (NEPRC) will introduce the best aptamers, via retroviral vectors, into macaque CD34+ve cells, transplant them into macaques, examine levels of gene marking in uninfected macaques, as well as test the efficacy of the aptamers to protect CD4 +ve T cells derived from marked CD34 +ve cells in vivo.