Determine the Transcriptional Activity of Non-integrated HIV-1 DNA

描述（由申请人提供）：感染人类免疫缺陷病毒（HIV）导致CD4 T细胞耗竭并导致免疫抑制状态。病毒的复制需要将其基因组稳定地整合到人类染色体中，但HIV感染的自然过程导致大量未整合的病毒DNA在体内积累。尽管在体外非整合病毒DNA的转录活性得到了明确的证明，但在了解这一早期过程的调控及其在HIV生命周期中的直接作用方面进展甚微。因此，非整合病毒DNA在HIV感染中的致病潜力尚不清楚。这项应用的长期目标是了解调节前整合转录的分子机制以及这一早期过程在病毒发病机制中的作用。本研究的具体目的是确定调节整合前转录的病毒DNA模板，以及HIV Tat参与整合前转录的启动。实现这些特定目标的实验方法是使用我们最近构建的HIV整合酶阴性病毒和一种新的指示细胞来表征整合前转录。预计从这些研究中获得的数据将有助于深入了解非整合病毒DNA能够产生病毒因子、增强病毒复制和促进病毒发病机制的情况。提出的研究将有助于了解非整合HIV DNA在疾病进展中的潜在致病作用。此外，在某些情况下，非整合病毒DNA可以作为疾病进展的临床标志物。

DESCRIPTION (provided by applicant): Infection by the Human Immunodeficiency Virus (HIV) causes depletion of CD4 T cells and leads to a state of immuno-suppression. The viral replication requires stable integration of its genome into the human chromosome, yet the natural process of HIV infection leads to the accumulation of a large quantity of unintegrated viral DNA in the body. In spite of clear demonstration of transcriptional activity from non-integrated viral DNA in vitro, little progress has been made in understanding regulation of this early process and its direct role in the HIV life cycle. As such, pathogenic potential of non- integrated viral DNA in HIV infection is not well understood. The long-term objectives of this application are to understand the molecular mechanisms that regulate pre- integration transcription and the contribution of this early process in viral pathogenesis. The specific aims of this proposal are to determine the viral DNA template regulating pre-integration transcription, and the involvement of HIV Tat in the initiation of pre- integration transcription. The experiment methods to achieve these specific aims are to use an HIV integrase negative virus and a novel indicator cell that we have recently constructed to characterize pre-integration transcription. It is expected that data acquired from these studies will make help to gain insight into circumstances where non-integrated viral DNA is capable of producing viral factors, enhancing viral replication and contributing to viral pathogenesis. The proposed research will help to understand potential pathogenic role of non-integrated HIV DNA in disease progression. Additionally, non-integrated viral DNA could serve as clinical markers for disease progression in certain situation.