Dunn Stephen J, Khan Imran H, Chan Ursula A, Scearce Robin L, Melara Claudia L, Paul Amber M, Sharma Vikram, Bih Fong-Yih, Holzmayer Tanya A, Luciw Paul A, Abo Arie
PPD Discovery, Inc., Menlo Park, CA 94025, USA.
Virology. 2004 Apr 10;321(2):260-73. doi: 10.1016/j.virol.2004.01.010.
Human immunodeficiency virus (HIV) drugs designed to interfere with obligatory utilization of certain host cell factors by virus are less likely to encounter development of resistant strains than drugs directed against viral components. Several cellular genes required for productive infection by HIV were identified by the use of genetic suppressor element (GSE) technology as potential targets for anti-HIV drug development. Fragmented cDNA libraries from various pools of human peripheral blood mononuclear cells (PBMC) were expressed in vitro in human immunodeficiency virus type 1 (HIV-1)-susceptible cell lines and subjected to genetic screens to identify GSEs that interfered with viral replication. After three rounds of selection, more than 15000 GSEs were sequenced, and the cognate genes were identified. The GSEs that inhibited the virus were derived from a diverse set of genes including cell surface receptors, cytokines, signaling proteins, transcription factors, as well as genes with unknown function. Approximately 2.5% of the identified genes were previously shown to play a role in the HIV-1 life cycle; this finding supports the biological relevance of the assay. GSEs were derived from the following 12 cell surface proteins: CXCR4, CCR4, CCR7, CD11C, CD44, CD47, CD68, CD69, CD74, CSF3R, GABBR1, and TNFR2. Requirement of some of these genes for viral infection was also investigated by using RNA interference (RNAi) technology; accordingly, 10 genes were implicated in early events of the viral life cycle, before viral DNA synthesis. Thus, these cell surface proteins represent novel targets for the development of therapeutics against HIV-1 infection and AIDS.
旨在干扰病毒对某些宿主细胞因子的必需利用的人类免疫缺陷病毒（HIV）药物，比针对病毒成分的药物更不容易出现耐药菌株的产生。通过使用遗传抑制元件（GSE）技术，确定了HIV有效感染所需的几个细胞基因，作为抗HIV药物开发的潜在靶点。来自人类外周血单核细胞（PBMC）不同库的片段化cDNA文库在1型人类免疫缺陷病毒（HIV-1）易感细胞系中进行体外表达，并进行遗传筛选，以鉴定干扰病毒复制的GSE。经过三轮筛选，对15000多个GSE进行了测序，并确定了相关基因。抑制病毒的GSE来自多种基因，包括细胞表面受体、细胞因子、信号蛋白、转录因子以及功能未知的基因。先前已证明约2.5%的已鉴定基因在HIV-1生命周期中发挥作用；这一发现支持了该检测方法的生物学相关性。GSE来自以下12种细胞表面蛋白：CXCR4、CCR4、CCR7、CD11C、CD44、CD47、CD68、CD69、CD74、CSF3R、GABBR1和TNFR2。还通过使用RNA干扰（RNAi）技术研究了其中一些基因对病毒感染的需求；因此，有10个基因与病毒生命周期中病毒DNA合成之前的早期事件有关。因此，这些细胞表面蛋白代表了开发抗HIV-1感染和艾滋病治疗药物的新靶点。
