在一项新的研究中,来自美国怀特海德研究所、拉根研究所和布罗德研究所的研究人员利用CRISPR-Cas9基因编辑技术鉴定出三个有望用于治疗HIV感染的新靶标。他们描述了如何利用CRISPR筛选HIV感染但不是细胞存活所必需的人基因的方法鉴定出5个基因---它们的三个在早前的利用RNA干扰(RNAi)的研究中并未被鉴定出。他们的方法也能够被用来鉴定出针对其他的病毒性病原体的治疗性靶标。相关研究结果于2016年12月19日在线发表在Nature Genetics期刊上,论文标题为“A genome-wide CRISPR screen identifies a restricted set of HIV host dependency factors”。论文通信作者为David M Sabatini、Eric S Lander、Nir Hacohen和Bruce D Walker。
Park解释道,“在我们的细胞系中,ALCAM是细胞间粘附所必需的,从而允许病毒更加高效地从一个细胞转移到下一个细胞。事实上,我们发现人工诱导缺乏ALCAM的细胞聚集会恢复HIV的细胞间传播。还需开展进一步的研究来探究靶向这些基因是否对人体是有毒性的。然而,即便系统性抑制会有毒性作用,仅在CD4阳性T细胞或它们的前体细胞中选择性地靶向这些基因可能会避免这些毒性,不过应注意到基因疗法仍然是一种充满挑战性的潜在高成本的治疗方法。” A genome-wide CRISPR screen identifies a restricted set of HIV host dependency factors
Ryan J Park, Tim Wang, Dylan Koundakjian, Judd F Hultquist, Pedro Lamothe-Molina, Blandine Monel, Kathrin Schumann, Haiyan Yu, Kevin M Krupzcak, Wilfredo Garcia-Beltran, Alicja Piechocka-Trocha, Nevan J Krogan, Alexander Marson, David M Sabatini, Eric S Lander, Nir Hacohen & Bruce D Walker
doi:10.1038/ng.3741
(生物谷 Bioon.com) 作者: marine0425030 时间: 2017-1-2 00:21
abstract:
Host proteins are essential for HIV entry and replication and can be important nonviral therapeutic targets. Large-scale RNA interference (RNAi)-based screens have identified nearly a thousand candidate host factors, but there is little agreement among studies and few factors have been validated. Here we demonstrate that a genome-wide CRISPR-based screen identifies host factors in a physiologically relevant cell system. We identify five factors, including the HIV co-receptors CD4 and CCR5, that are required for HIV infection yet are dispensable for cellular proliferation and viability. Tyrosylprotein sulfotransferase 2 (TPST2) and solute carrier family 35 member B2 (SLC35B2) function in a common pathway to sulfate CCR5 on extracellular tyrosine residues, facilitating CCR5 recognition by the HIV envelope. Activated leukocyte cell adhesion molecule (ALCAM) mediates cell aggregation, which is required for cell-to-cell HIV transmission. We validated these pathways in primary human CD4+ T cells through Cas9-mediated knockout and antibody blockade. Our findings indicate that HIV infection and replication rely on a limited set of host-dispensable genes and suggest that these pathways can be studied for therapeutic intervention.