INHIBITION OF SHIV REPLICATION BY HIV-SPECIFIC APTAMERS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 尽管高效抗逆转录病毒疗法(HAART)在抑制艾滋病毒感染者体内的病毒复制方面取得了巨大成功，但越来越明显的是，开发补充疗法，尤其是针对艾滋病毒疾病的基因疗法，有令人信服的理由。最近的实验表明，RT-特异性适配子对HIV-1和RT-SHV复制具有相当强的抑制作用。作为抑制艾滋病毒复制的一种方式，适体具有一些独特的优势，包括靶向逆转录病毒生命周期的多种元件的能力，它们对逃逸病毒出现的相对抵抗力，以及它们在抑制艾滋病毒复制方面的观察到的效力。在用表达HIV-1特异性适配子的逆转录病毒载体转导细胞系后，我们观察到Shiv RT复制显著受到抑制。稳定表达适配子的逆转录病毒产生细胞系已经建立并用于转导转化的细胞系。对这些适配子在细胞系中抑制病毒复制的能力的分析没有显示出明显的抑制作用。然后我们确定U6启动子发生了缺失，发现U6启动子在反义方向上含有隐蔽的供体/受体剪接位点。这些研究应该会产生关于基于适体的干细胞基因疗法治疗艾滋病的有效性和安全性的重要信息，并最终促进在艾滋病毒感染者中开展类似的试验。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Despite the dramatic success of highly active antiretroviral therapy (HAART) in inhibiting viral replication in HIV-infected subjects, it is increasingly clear that there is a compelling rationale for the development of complementary therapies, most notably genetic therapies for HIV disease. Recent experiments have demonstrated quite potent inhibition of HIV-1 and RT-SHIV replication by RT-specific aptamers. Aptamers have a number of distinctive advantages as a modality to inhibit HIV replication, including the ability to target multiple elements of the retroviral life cycle, their relative resistance to the emergence of escape viruses, and their observed potency in inhibiting HIV replication. Following transduction of cell lines with retroviral vectors expressing aptamers specific for HIV-1, we observed significant inhibition of SHIV RT replication. Stable retroviral producer cell lines expressing aptamers have been generated and used to transduce transformed cell lines. Analysis of the ability of these aptamers to inhibit viral replication in cell lines did not reveal significant inhibition. We then determined that a deletion had occurred in the U6 promoter, which was found to contain a cryptic donor/acceptor splice site in the antisense orientation. These studies should yield important information regarding the efficacy and safety of aptamer-based stem cell gene therapy for AIDS and ultimately facilitate the development of similar trials in HIV-infected people.