Cisneros William J, Walter Miriam, Soliman Shimaa H A, Simons Lacy M, Cornish Daphne, Halle Ariel W, Kim Eun-Young, Wolinsky Steven M, Shilatifard Ali, Hultquist Judd F
Division of Infectious Diseases, Department of Medicine, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
Center for Pathogen Genomics and Microbial Evolution, Havey Institute for Global Health, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
bioRxiv. 2024 Mar 1:2024.03.01.582881. doi: 10.1101/2024.03.01.582881.
The persistence of HIV-1 in long-lived latent reservoirs during suppressive antiretroviral therapy (ART) remains one of the principal barriers to a functional cure. Blocks to transcriptional elongation play a central role in maintaining the latent state, and several latency reversal strategies focus on the release of positive transcription elongation factor b (P-TEFb) from sequestration by negative regulatory complexes, such as the 7SK complex and BRD4. Another major cellular reservoir of P-TEFb is in Super Elongation Complexes (SECs), which play broad regulatory roles in host gene expression. Still, it is unknown if the release of P-TEFb from SECs is a viable latency reversal strategy. Here, we demonstrate that the SEC is not required for HIV-1 replication in primary CD4+ T cells and that a small molecular inhibitor of the P-TEFb/SEC interaction (termed KL-2) increases viral transcription. KL-2 acts synergistically with other latency reversing agents (LRAs) to reactivate viral transcription in several cell line models of latency in a manner that is, at least in part, dependent on the viral Tat protein. Finally, we demonstrate that KL-2 enhances viral reactivation in peripheral blood mononuclear cells (PBMCs) from people living with HIV on suppressive ART, most notably in combination with inhibitor of apoptosis protein antagonists (IAPi). Taken together, these results suggest that the release of P-TEFb from cellular SECs may be a novel route for HIV-1 latency reactivation.
在抗逆转录病毒疗法(ART)抑制病毒复制期间,HIV-1在长期存在的潜伏库中的持续存在仍然是实现功能性治愈的主要障碍之一。转录延伸受阻在维持潜伏状态中起核心作用,几种潜伏逆转策略聚焦于将正性转录延伸因子b(P-TEFb)从诸如7SK复合体和BRD4等负调控复合体的隔离中释放出来。P-TEFb的另一个主要细胞储存库存在于超级延伸复合体(SEC)中,其在宿主基因表达中发挥广泛的调控作用。然而,P-TEFb从SEC中释放是否是一种可行的潜伏逆转策略尚不清楚。在此,我们证明在原代CD4+ T细胞中HIV-1复制不需要SEC,并且一种P-TEFb/SEC相互作用的小分子抑制剂(称为KL-2)可增加病毒转录。在几种潜伏细胞系模型中,KL-2与其他潜伏逆转剂(LRA)协同作用以重新激活病毒转录,其方式至少部分依赖于病毒Tat蛋白。最后,我们证明KL-2可增强接受抑制性ART治疗的HIV感染者外周血单核细胞(PBMC)中的病毒重新激活,最显著的是与凋亡蛋白拮抗剂抑制剂(IAPi)联合使用时。综上所述,这些结果表明P-TEFb从细胞SEC中释放可能是HIV-1潜伏逆转的一条新途径。