INHIBITION OF SHIV REPLICATION BY HIV-SPECIFIC APTAMERS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 尽管高效抗逆转录病毒疗法（HAART）在抑制HIV感染者的病毒复制方面取得了巨大成功，但越来越清楚的是，开发补充疗法，特别是HIV疾病的基因疗法，有着令人信服的理由。 最近的实验表明，RT特异性适体对HIV-1和RT-SHIV复制具有相当强的抑制作用。 适体作为抑制HIV复制的方式具有许多独特的优势，包括靶向逆转录病毒生命周期的多个元件的能力，它们对逃逸病毒出现的相对抗性，以及它们在抑制HIV复制中观察到的效力。 用表达HIV-1特异性适体的逆转录病毒载体转导细胞系后，我们观察到SHIV RT复制的显著抑制。 已经产生了表达适体的稳定的逆转录病毒生产细胞系，并将其用于逆转录转化的细胞系。对这些适体抑制细胞系中病毒复制的能力的分析没有显示出显著的抑制。 然后，我们确定在U6启动子中发生了缺失，发现其在反义方向上含有隐蔽的供体/受体剪接位点。 这些研究应产生重要的信息，对艾滋病的适体为基础的干细胞基因疗法的疗效和安全性，并最终促进艾滋病毒感染者的类似试验的发展。

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.