Proteins Mediating Interaction of HIV-1 and JCV in CNS

描述（由申请人提供）：该项目的总体目标是阐明HIV-1感染影响脑胶质细胞中JC病毒激活的机制。JC病毒（JCV）是神经退行性疾病进行性多灶性白质脑病（PML）的病因。通常情况下，JCV在非免疫功能低下的人群中是潜伏的，但它在艾滋病患者的大脑中被激活。由于PML在艾滋病中的高发病率，我们假设HIV-1发挥的作用比免疫抑制单独发挥的作用更大。我们将利用近几年来取得的广泛成果，这些成果可分为两类。在一项研究中，我们证明了HIV-1的Tat蛋白在刺激JCV晚期基因转录和JCV DMA复制中的作用。另一方面，我们已经证明HIV-1感染改变了免疫调节剂（包括TGF-01）在中枢神经系统中的产生和信号转导途径。我们打算确定TGF-31的Smad核效应物如何在JCV和PCNA启动子序列上与Tat及其细胞伴侣蛋白Pura和Cyclin T1/ Cdk9相互作用。我们的新双染色质免疫沉淀方法将与JCV DMA复制和基因转录的功能研究结合使用。我们将采用微阵列来鉴定受暴露于hiv -1感染细胞产生的细胞因子调节的神经胶质细胞中的基因。我们将采用转基因小鼠模型来验证hiv -1感染细胞产生的因子可以影响中枢神经系统中的JCV启动子并在PML发展的早期阶段发挥作用的假设。来自曼哈顿HIV脑库的组织将用于确定TGF-P1或其核效应物Smad3、Smad4或Fasti是否在PML病变的特定细胞中定位或激活。结果将有助于阐明大脑中JCV的激活途径，并将有助于靶向特定的分子相互作用进行治疗。

DESCRIPTION (provided by applicant): The overall goal of this project is to elucidate the mechanisms by which activation of JC virus in glial cells of the brain is influenced by HIV-1 infection. JC virus (JCV) is the etiologic agent of the neurodegenerative disease, progressive multifocal leukoencephalopathy (PML). JCV is normally latent in non-immunocompromised people, but it is activated in brains of individuals with AIDS. Because of the high incidence of PML in AIDS, we have hypothesized that HIV-1 plays a role greater than that expected of immunosuppression alone. We shall capitalize upon our extensive results of the last few years, which can be grouped in two categories. In one we have demonstrated the role of the Tat protein of HIV-1 in stimulating JCV late gene transcription and JCV DMA replication. In the other we have demonstrated that HIV-1 infection alters pathways of production and signal transduction of immunomodulators, including TGF-01, in the CNS. We propose to determine how the Smad nuclear effectors of TGF-31 interact with Tat and its cellular partner proteins Pura and Cyclin T1/ Cdk9 at JCV and PCNA promoter sequences. Our new double chromatin immunoprecipitation method will be employed in conjunction with functional studies on JCV DMA replication and gene transcription. We shall employ a microarray to identify genes in glial cells regulated by exposure to cytokines produced by HIV-1-infected cells. We shall employ a transgenic mouse model to test the hypothesis that factors produced by HIV-1-infected cells can influence JCV promoters in the CNS and play a role in early steps of PML development. Tissue from the Manhattan HIV Brain Bank will be used to determine whether TGF-P1 or its nuclear effectors, Smad3, Smad4 or Fasti, are localized or activated in specific cells of PML lesions. Results will help elucidate pathways of activation of JCV in the brain and will help target particular molecular interactions for therapy.