Department of Biochemistry, Erasmus MC University Medical Center Rotterdam, PO Box 2040, 3000 CA Rotterdam, Netherlands.
Westerdijk Fungal Biodiversity Institute, Utrecht, Netherlands.
Sci Adv. 2020 Aug 12;6(33):eaba6617. doi: 10.1126/sciadv.aba6617. eCollection 2020 Aug.
A leading pharmacological strategy toward HIV cure requires "shock" or activation of HIV gene expression in latently infected cells with latency reversal agents (LRAs) followed by their subsequent clearance. In a screen for novel LRAs, we used fungal secondary metabolites as a source of bioactive molecules. Using orthogonal mass spectrometry (MS) coupled to latency reversal bioassays, we identified gliotoxin (GTX) as a novel LRA. GTX significantly induced HIV-1 gene expression in latent ex vivo infected primary cells and in CD4 T cells from all aviremic HIV-1 participants. RNA sequencing identified 7SK RNA, the scaffold of the positive transcription elongation factor b (P-TEFb) inhibitory 7SK small nuclear ribonucleoprotein (snRNP) complex, to be significantly reduced upon GTX treatment of CD4 T cells. GTX directly disrupted 7SK snRNP by targeting La-related protein 7 (LARP7), releasing active P-TEFb, which phosphorylated RNA polymerase II (Pol II) C-terminal domain (CTD), inducing HIV transcription.
一种主要的抗 HIV 治愈策略需要使用潜伏逆转剂（LRAs）对潜伏感染细胞进行“冲击”或激活 HIV 基因表达，然后清除它们。在寻找新型 LRA 的筛选中，我们使用真菌次级代谢产物作为生物活性分子的来源。通过正交质谱（MS）与潜伏逆转生物测定相结合，我们鉴定出了Gliotoxin（GTX）是一种新型的 LRA。GTX 可显著诱导潜伏感染的原代细胞和所有 HIV-1 无病毒血症参与者的 CD4 T 细胞中的 HIV-1 基因表达。RNA 测序鉴定出 7SK RNA，即正转录延伸因子 b（P-TEFb）抑制性 7SK 小核核糖核蛋白（snRNP）复合物的支架，在 GTX 处理 CD4 T 细胞后显著减少。GTX 通过靶向 La 相关蛋白 7（LARP7）直接破坏 7SK snRNP，释放活性 P-TEFb，磷酸化 RNA 聚合酶 II（Pol II）C 末端结构域（CTD），诱导 HIV 转录。
