Blocking HIV with aptamers targeted to viral components

描述（由申请人提供）：HIV复制的不完全抑制和随之产生的耐药性继续损害艾滋病治疗。潜在的问题包括功效、不粘附和感染的细胞库。虽然靶向进入、整合或转录的新药将改善一些问题，但实现更完整的解决方案需要转向新的补充疗法。在几种可用于HIV-1的有效基因治疗方法中，靶向HIV-1的RNA适体是有效的。已知适体具有特异性、高亲和力、稳定性和无免疫原性。最近的工作提供了令人信服的证据表明，针对HIV-1的细胞内适体可以强烈抑制病毒复制。赋予对此类适体的抗性的突变已导致病毒适应性的丧失。因此，存在迄今未利用的机会来开发具有独特特异性（降低毒性）的抗HIV剂，其将更完全地抑制HIV-1复制，阻碍或减缓抗性并消除不粘附。为此，提出了以下涉及各组织/研究人员的合作研究方案。1.德克萨斯州奥斯汀市（Austin，TX）与Andy Ellington（德克萨斯大学奥斯汀分校）合作，将进行针对HIV-1靶标的RNA适体的高通量选择，鉴定紧密结合物，确定并进一步优化体外功效。 2. Vinayaka Prasad（AECOM）将使用来自Accacia的预选适体来确定抑制HIV/SHIV复制的功效，确定抑制机制并选择适体抗性。 他将为Accacia提供抗性蛋白，用于开发第二代适体。3. Paul约翰逊（NEPRC）将通过逆转录病毒载体将最佳适体引入猕猴CD 34 +ve细胞，将其移植到猕猴体内，检查未感染猕猴的基因标记水平，并测试适体在体内保护来自标记CD 34 +ve细胞的CD 4 +ve T细胞的功效。

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.