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子介导其感染周期。正因为如此，一种破坏HIV-1蛋白与其细胞辅因子之间基本相互作用的小分子可以作为抗hiv药物。因此，细胞辅助因子的鉴定是开发新型抗hiv药物的必要的第一步。这项拨款申请旨在鉴定我们实验室在转录谱研究中鉴定的一组54个基因中的新型HIV-1辅因子。这些基因在活化的CD4+ T细胞和分化的巨噬细胞中均上调，其上调依赖于Cyclin T1蛋白。Cyclin T1是HIV-1 Tat蛋白的直接靶点，它介导整合前病毒的RNAP II转录。虽然我们的基因列表中的大多数编码蛋白尚未被评估在HIV-1感染中的作用，但在文献中已经报道了10个（54个）在HIV-1感染周期中发挥作用。在随机生成的54组在CD4+ T细胞和巨噬细胞中表达的基因中，我们观察到只有2或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HIV-1复制的蛋白质中被过度代表（p值<0.00021）。这一统计分析表明，我们的名单极有可能包含新的病毒辅助因子。为了确定新的HIV-1辅助因子，我们提出了两个具体目标。用我们列表中的一个基因酪蛋白激酶1 γ 1 （CSNK1G1）进行的初步实验表明，这种细胞蛋白是影响HIV-1病毒粒子感染性的辅助因子。Specific Aim #1提出研究CSNK1G1在HIV-1复制中的作用，并确定细胞因子是否影响病毒粒子的感染性。特异性目标#2建议进行shRNA筛选，以确定我们的54个周期蛋白t1依赖基因中的哪一个在HIV-1复制中起作用。这项研究的完成可能会确定新的靶点，这可能是开发新型抗HIV疗法的基础。鉴于HIV-1能够获得对当前抗病毒药物的耐药性以及这些药物对许多患者的毒性，基础研究面临的一个持续挑战是开发新的抗hiv药物。公共卫生相关性：HIV-1蛋白必需的细胞辅助因子可作为抗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。本申请中提出的研究将调查该基因集是否确实包含可以作为抗hiv药物靶点的新型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.