Mutations Located outside the Integrase Gene Can Confer Resistance to HIV-1 Integrase Strand Transfer Inhibitors.
Mutations Located outside the Integrase Gene Can Confer Resistance to HIV-1 Integrase Strand Transfer Inhibitors.
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DOI：
10.1128/mbio.00922-17
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发表时间：
2017-09-26
期刊：
影响因子：
6.4
通讯作者：
Delelis O
中科院分区：
文献类型：
--
作者：
Malet I;Subra F;Charpentier C;Collin G;Descamps D;Calvez V;Marcelin AG;Delelis O
对整合酶链转移抑制剂拉替拉韦和埃替拉韦的耐药性通常是由于整合酶基因中已明确的突变所致。然而，对于多替拉韦治疗失败的患者来说，情况不太清楚。此外，大多数选择多替拉韦耐药性的体外实验导致整合酶基因很少发生突变。我们采用突破性方法进行了体外多替拉韦耐药性选择实验。首先，MT4细胞被人类免疫缺陷病毒1型（HIV-1）莱感染。整合到宿主细胞基因组中后，洗涤细胞以去除未结合的病毒，并向细胞培养基中添加 500 nM 多替拉韦。在整个选择过程中保持药物的这种高浓度。第 80 天时，我们检测到一种对多替拉韦、拉替拉韦和艾维拉韦高度耐药的病毒，但仍对齐多夫定敏感。病毒测序显示整合酶基因没有突变，但突出显示了五个突变的出现，全部位于 nef 区域，其中四个聚集在 3' 多嘌呤区 (PPT)。位于整合酶基因外部的多替拉韦在体外选择的突变可以赋予对所有整合酶抑制剂的高水平耐药性。因此，HIV-1 可以使用另一种机制，通过选择 3' PPT 区域的突变来产生对整合酶抑制剂的耐药性。需要进一步的研究来确定这些突变在多大程度上可以解释整合酶抑制剂治疗期间的病毒学失败。整合酶链转移抑制剂（INSTI）越来越多地用作一线药物和挽救治疗，因为它们对初治和有治疗经验的患者毒性低且疗效高。到目前为止，通过 INSTI 暴露选择的耐药突变要么在患者中得到描述，要么在体外选择并涉及整合酶基因。拉替拉韦、埃替拉韦或多替拉韦暴露选择的大多数突变位于整合酶基因催化位点内部，但多替拉韦也选择了整合酶基因催化位点外部的突变。在使用多替拉韦进行体外选择后，我们首次报告了一种在 HIV-1 整合酶基因外具有选定突变的病毒，该突变对目前用于治疗患者的所有整合酶抑制剂（例如拉替拉韦、埃替拉韦和多替拉韦）产生耐药性。我们的观察结果可以解释为什么一些导致接受多替拉韦治疗的患者病毒学失败的病毒没有表现出整合酶基因突变。
Resistance to the integrase strand transfer inhibitors raltegravir and elvitegravir is often due to well-identified mutations in the integrase gene. However, the situation is less clear for patients who fail dolutegravir treatment. Furthermore, most in vitro experiments to select resistance to dolutegravir have resulted in few mutations of the integrase gene. We performed an in vitro dolutegravir resistance selection experiment by using a breakthrough method. First, MT4 cells were infected with human immunodeficiency virus type 1 (HIV-1) Lai. After integration into the host cell genome, cells were washed to remove unbound virus and 500 nM dolutegravir was added to the cell medium. This high concentration of the drug was maintained throughout selection. At day 80, we detected a virus highly resistant to dolutegravir, raltegravir, and elvitegravir that remained susceptible to zidovudine. Sequencing of the virus showed no mutations in the integrase gene but highlighted the emergence of five mutations, all located in the nef region, of which four were clustered in the 3′ polypurine tract (PPT). Mutations selected in vitro by dolutegravir, located outside the integrase gene, can confer a high level of resistance to all integrase inhibitors. Thus, HIV-1 can use an alternative mechanism to develop resistance to integrase inhibitors by selecting mutations in the 3′ PPT region. Further studies are required to determine to what extent these mutations may explain virological failure during integrase inhibitor therapy. Integrase strand transfer inhibitors (INSTIs) are increasingly used both as first-line drugs and in rescue therapy because of their low toxicity and high efficacy in both treatment-naive and treatment-experienced patients. Until now, resistance mutations selected by INSTI exposure have either been described in patients or selected in vitro and involve the integrase gene. Most mutations selected by raltegravir, elvitegravir, or dolutegravir exposure are located inside the catalytic site of the integrase gene, but mutations outside the catalytic site of the integrase gene have also been selected with dolutegravir. Following in vitro selection with dolutegravir, we report, for the first time, a virus with selected mutations outside the HIV-1 integrase gene that confer resistance to all integrase inhibitors currently used to treat patients, such as raltegravir, elvitegravir, and dolutegravir. Our observation may explain why some viruses responsible for virological failure in patients treated with dolutegravir did not show mutations in the integrase gene.