Experimental Retrovirology Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan.
Sci Adv. 2023 Jul 14;9(28):eadg2955. doi: 10.1126/sciadv.adg2955. Epub 2023 Jul 12.
Nuclear localization signal (NLS) of HIV-1 integrase (IN) is implicated in nuclear import of HIV-1 preintegration complex (PIC). Here, we established a multiclass drug-resistant HIV-1 variant (HIV) by consecutively exposing an HIV-1 variant to various antiretroviral agents including IN strand transfer inhibitors (INSTIs). HIV was extremely susceptible to a previously reported HIV-1 protease inhibitor, GRL-142, with IC of 130 femtomolar. When cells were exposed to HIV IN-containing recombinant HIV in the presence of GRL-142, significant decrease of unintegrated 2-LTR circular cDNA was observed, suggesting that nuclear import of PIC was severely compromised by GRL-142. X-ray crystallographic analyses revealed that GRL-142 interacts with NLS's putative sequence (DQAEHLK) and sterically blocks the nuclear transport of GRL-142-bound HIV's PIC. Highly INSTI-resistant HIV-1 variants isolated from heavily INSTI-experienced patients proved to be susceptible to GRL-142, suggesting that NLS-targeting agents would serve as salvage therapy agents for highly INSTI-resistant variant-harboring individuals. The data should offer a new modality to block HIV-1 infectivity and replication and shed light on developing NLS inhibitors for AIDS therapy.
HIV-1 整合酶（IN）的核定位信号（NLS）被认为参与了 HIV-1 前整合复合物（PIC）的核内输入。在这里，我们通过连续暴露 HIV-1 变体于包括整合酶链转移抑制剂（INSTIs）在内的各种抗逆转录病毒药物，建立了一个多药耐药的 HIV-1 变体（HIV）。HIV 对先前报道的 HIV-1 蛋白酶抑制剂 GRL-142 极其敏感，IC 为 130 飞摩尔。当细胞在 GRL-142 的存在下暴露于含有 HIV IN 的重组 HIV 时，观察到未整合的 2-LTR 环状 cDNA 显著减少，这表明 GRL-142 严重损害了 PIC 的核内输入。X 射线晶体学分析表明，GRL-142 与 NLS 的假定序列（DQAEHLK）相互作用，并在空间上阻止 GRL-142 结合的 HIV 的 PIC 的核转运。从大量使用 INSTI 的患者中分离出的高度耐药的 HIV-1 变体对 GRL-142 敏感，这表明 NLS 靶向药物将作为高度耐药变体携带个体的挽救治疗药物。这些数据为阻断 HIV-1 感染和复制提供了一种新的方法，并为开发用于艾滋病治疗的 NLS 抑制剂提供了思路。
