Detection of treatment-resistant infectious HIV after genome-directed antiviral endonuclease therapy.
Detection of treatment-resistant infectious HIV after genome-directed antiviral endonuclease therapy.
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DOI：
10.1016/j.antiviral.2015.12.007
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发表时间：
2016-02
影响因子：
7.6
通讯作者：
Jerome KR
中科院分区：
文献类型：
--
作者：
De Silva Feelixge HS;Stone D;Pietz HL;Roychoudhury P;Greninger AL;Schiffer JT;Aubert M;Jerome KR
不可治愈的慢性病毒感染是全世界发病率和死亡率的主要原因。治愈持续性病毒感染的一种潜在方法是通过使用靶向核酸内切酶。然而，基于核酸内切酶的抗病毒疗法的一个潜在问题是出现治疗耐药性。在这里，我们首次检测到一种耐核酸内切酶的感染性病毒，这种病毒在抗病毒核酸内切酶治疗后出现频率很高。在测试HIV pol特异性锌指核酸酶（ZFN）单独或与三个引物修复核酸外切酶2（Trex 2）组合的活性时，我们鉴定了一种治疗抗性和感染性突变病毒，其来源于ZFN介导的逆转录酶（RT）破坏。虽然HIV蛋白酶、RT和整合酶的基因破坏可以抑制病毒复制，但是在RT的拇指结构域中插入一个偶然的单个氨基酸产生了可以积极复制的病毒。耐核酸内切酶病毒可以在原代CD 4 + T细胞中复制，但仍然对抗逆转录病毒RT抑制剂治疗敏感。当二级ZFN衍生的突变被引入突变病毒的RT或整合酶结构域时，复制可以被消除。我们的观察表明，在基于核酸内切酶的抗病毒治疗期间应谨慎;然而，联合核酸内切酶治疗可能会防止耐药性的出现。
Incurable chronic viral infections are a major cause of morbidity and mortality worldwide. One potential approach to cure persistent viral infections is via the use of targeted endonucleases. Nevertheless, a potential concern for endonuclease-based antiviral therapies is the emergence of treatment resistance. Here we detect for the first time an endonuclease-resistant infectious virus that is found with high frequency after antiviral endonuclease therapy. While testing the activity of HIV pol-specific zinc finger nucleases (ZFNs) alone or in combination with three prime repair exonuclease 2 (Trex2), we identified a treatment-resistant and infectious mutant virus that was derived from a ZFN-mediated disruption of reverse transcriptase (RT). Although gene disruption of HIV protease, RT and integrase could inhibit viral replication, a chance single amino acid insertion within the thumb domain of RT produced a virus that could actively replicate. The endonuclease-resistant virus could replicate in primary CD4+ T cells, but remained susceptible to treatment with antiretroviral RT inhibitors. When secondary ZFN-derived mutations were introduced into the mutant virus’s RT or integrase domains, replication could be abolished. Our observations suggest that caution should be exercised during endonuclease-based antiviral therapies; however, combination endonuclease therapies may prevent the emergence of resistance.