Sequential LASER ART and CRISPR Treatments Eliminate HIV-1 in a Subset of Infected Humanized Mice
Sequential LASER ART and CRISPR Treatments Eliminate HIV-1 in a Subset of Infected Humanized Mice
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DOI：
10.1038/s41467-019-10366-y
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发表时间：
2019-07-02
影响因子：
16.6
通讯作者：
Gendelman, Howard E.
中科院分区：
文献类型：
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作者：
Dash, Prasanta K.;Kaminski, Rafal;Gendelman, Howard E.
消除HIV-1需要从感染的细胞和组织中清除和去除整合的前病毒DNA。在此，连续长效缓释抗病毒疗法（LASER ART）和CRISPR-Cas9证明了在HIV-1感染的人源化小鼠中潜伏感染性储库中的病毒清除。HIV-1亚基因组DNA片段，跨越长末端重复序列和Gag基因，在体内被切除，导致整合的前病毒DNA的消除;通过巢式和数字液滴PCR以及RNAscope测试，在血液、淋巴组织、骨髓和脑中未检测到病毒。未检测到CRISPR-Cas9介导的脱靶效应。将人免疫细胞从双重处理的无病毒动物连续转移到未感染的人源化小鼠不能产生感染性子代病毒。相比之下，HIV-1在单独的LASER ART或CRISPR-Cas9治疗后很容易被检测到。这些数据提供了概念验证，即永久消除病毒是可能的。
Elimination of HIV-1 requires clearance and removal of integrated proviral DNA from infected cells and tissues. Here, sequential long-acting slow-effective release antiviral therapy (LASER ART) and CRISPR-Cas9 demonstrate viral clearance in latent infectious reservoirs in HIV-1 infected humanized mice. HIV-1 subgenomic DNA fragments, spanning the long terminal repeats and the Gag gene, are excised in vivo, resulting in elimination of integrated proviral DNA; virus is not detected in blood, lymphoid tissue, bone marrow and brain by nested and digital-droplet PCR as well as RNAscope tests. No CRISPR-Cas9 mediated off-target effects are detected. Adoptive transfer of human immunocytes from dual treated, virus-free animals to uninfected humanized mice fails to produce infectious progeny virus. In contrast, HIV-1 is readily detected following sole LASER ART or CRISPR-Cas9 treatment. These data provide proof-of-concept that permanent viral elimination is possible.