Aaron Diamond AIDS Research Center, Columbia University Irving Medical Center, New York, NY, 10032, USA.
Department of Microbiology and Immunology, Columbia University Irving Medical Center, New York, NY, 10032, USA.
Sci Rep. 2020 Mar 24;10(1):5350. doi: 10.1038/s41598-020-62375-3.
The major barrier to a HIV-1 cure is the persistence of latent genomes despite treatment with antiretrovirals. To investigate host factors which promote HIV-1 latency, we conducted a genome-wide functional knockout screen using CRISPR-Cas9 in a HIV-1 latency cell line model. This screen identified IWS1, POLE3, POLR1B, PSMD1, and TGM2 as potential regulators of HIV-1 latency, of which PSMD1 and TMG2 could be confirmed pharmacologically. Further investigation of PSMD1 revealed that an interacting enzyme, the deubiquitinase UCH37, was also involved in HIV-1 latency. We therefore conducted a comprehensive evaluation of the deubiquitinase family by gene knockout, identifying several deubiquitinases, UCH37, USP14, OTULIN, and USP5 as possible HIV-1 latency regulators. A specific inhibitor of USP14, IU1, reversed HIV-1 latency and displayed synergistic effects with other latency reversal agents. IU1 caused degradation of TDP-43, a negative regulator of HIV-1 transcription. Collectively, this study is the first comprehensive evaluation of deubiquitinases in HIV-1 latency and establishes that they may hold a critical role.
HIV-1 治愈的主要障碍是尽管使用抗逆转录病毒药物治疗，但潜伏基因组仍然存在。为了研究促进 HIV-1 潜伏的宿主因素，我们使用 CRISPR-Cas9 在 HIV-1 潜伏细胞系模型中进行了全基因组功能敲除筛选。该筛选鉴定出 IWS1、POLE3、POLR1B、PSMD1 和 TGM2 作为 HIV-1 潜伏的潜在调节剂，其中 PSMD1 和 TGM2 可以通过药理学方法得到证实。对 PSMD1 的进一步研究表明，一种相互作用的酶，去泛素酶 UCH37，也参与了 HIV-1 的潜伏。因此，我们通过基因敲除对去泛素酶家族进行了全面评估，鉴定出几种去泛素酶，UCH37、USP14、OTULIN 和 USP5，作为可能的 HIV-1 潜伏调节剂。USP14 的特异性抑制剂 IU1 逆转了 HIV-1 潜伏，并与其他潜伏逆转剂显示出协同作用。IU1 导致 HIV-1 转录的负调节剂 TDP-43 的降解。总的来说，这项研究是对 HIV-1 潜伏中去泛素酶的首次全面评估，并确立了它们可能具有关键作用。
