Identification of novel HIV-1 co-factors

描述(由申请人提供)：HIV-1的复制依赖于细胞辅助因素来调节其感染周期。正因为如此，一种破坏HIV-1蛋白及其细胞辅助因子之间基本相互作用的小分子可以起到抗HIV药物的作用。因此，细胞共因子的鉴定是开发新型抗艾滋病毒药物的必要的第一步。这项拨款申请建议在我们实验室在转录图谱研究中确定的一组54个基因中识别新的HIV-1辅助因子。这些基因在活化的CD4+T细胞和分化的巨噬细胞中都上调，并且它们的上调依赖于Cyclin T1蛋白。Cyclin T1是HIV-1Tat蛋白的直接靶标，它介导整合的前病毒的RNAP II转录。虽然我们的基因列表中的大多数编码蛋白还没有被评估在HIV-1感染中的作用，但文献中已经报道了10个(54个中)在HIV-1感染循环中具有作用。在随机生成的54个基因中，同时在CD4+T细胞和巨噬细胞中表达，我们观察到只有2到3个基因(平均2.4个)与HIV-1有关。因此，我们的54个基因在参与艾滋病毒-1复制的蛋白质中过度表达(p值为&lt；0.00021)。这项统计分析认为，我们的清单极有可能包含新的病毒辅助因子。为了识别新的HIV-1辅助因子，我们提出了两个特定的目标。对我们列表中的一个基因--酪蛋白激酶1-伽马1(CSNK1G1)进行的初步实验表明，这种细胞蛋白是影响HIV-1病毒粒子传染性的辅助因素。具体目标#1建议调查CSNK1G1在HIV-1复制中的作用，并确定细胞因素是否影响病毒粒子的传染性。具体目标#2建议进行shRNA筛选，以确定我们的54个Cyclin T1依赖基因中的哪些具有复制HIV-1的作用。这项拟议研究的完成可能会确定新的靶点，这些靶点可以成为开发新型抗艾滋病毒疗法的基础。鉴于HIV-1对当前的抗病毒药物产生抗药性的能力，以及这些药物对许多患者的毒性，基础研究的一个持续挑战是开发新型抗艾滋病毒药物。与公共卫生相关：HIV-1蛋白所必需的细胞辅助因子可能是抗HIV药物的基础。我们已经确定了一组54个细胞基因，它们可能包含新的HIV-1辅助因子。在这项申请中提出的研究将调查该基因集是否确实包含可作为抗HIV药物靶点的新的HIV-1辅助因子。

DESCRIPTION (provided by applicant): HIV-1 replication is dependent upon cellular co-factors to mediate its infectious cycle. Because of this, a small molecule that disrupts an essential interaction between an HIV-1 protein and its cellular co-factor can act as an anti-HIV drug. The identification of cellular co-factors is therefore a necessary first step in the development of novel anti-HIV drugs. This grant application proposes to identify novel HIV-1 co-factors in a set of 54 genes that our laboratory identified in a transcriptional profiling study. These genes are up-regulated in both activated CD4+ T cells and differentiated macrophages, and their up-regulation is dependent upon the Cyclin T1 protein. Cyclin T1 is a direct target of the HIV-1 Tat protein and it mediates RNAP II transcription of the integrated provirus. Although most of the encoded proteins in our gene list have not been evaluated for a role in HIV-1 infection, 10 (of 54) have been reported in the literature as having a role in the HIV-1 infectious cycle. In randomly generated sets of 54 genes that are expressed in both CD4+ T cells and macrophages, we observed that only 2 or 3 genes (average 2.4) have links to HIV-1. Thus, our set of 54 genes is over-represented in proteins involved in HIV-1 replication (p value of <0.00021). This statistical analysis argues that our list is highly likely to contain novel viral co-factors. To identify novel HIV-1 co-factors, we propose two Specific Aims. Preliminary experiments with one gene from our list, Casein kinase 1 gamma 1 (CSNK1G1), suggests that this cellular protein is a co-factor that affects HIV-1 virion infectivity. Specific Aim #1 proposes to investigate the role of CSNK1G1 in HIV-1 replication and determine if the cellular factor affects virion infectivity. Specific Aim #2 proposes to conduct a shRNA screen to determine which of our set of 54 Cyclin T1-dependent genes have a role HIV-1 replication. Completion of the proposed research is likely to identify new targets that can be the basis for the development of novel anti- HIV therapeutics. Given the ability of HIV-1 to acquire resistance to current anti-viral drugs and the toxicities of these drugs for many patients, an ongoing challenge for basic research is the development of novel anti-HIV drugs. PUBLIC HEALTH RELEVANCE: Cellular co-factors that are necessary for HIV-1 protein can be the basis of anti-HIV drugs. We have identified a set of 54 cellular genes that are likely to contain new HIV-1 co-factors. The research proposed in this application will investigate if this gene set does indeed contain novel HIV-1 co-factors that can be targets for anti-HIV drugs.