Combined noncanonical NF-κB agonism and targeted BET bromodomain inhibition reverse HIV latency ex vivo.
Combined noncanonical NF-κB agonism and targeted BET bromodomain inhibition reverse HIV latency ex vivo.
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DOI：
10.1172/jci157281
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发表时间：
2022-04-15
影响因子：
15.9
通讯作者：
Margolis, David M.
中科院分区：
文献类型：
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作者：
Falcinelli, Shane D.;Peterson, Jackson J.;Turner, Anne-Marie W.;Irlbeck, David;Read, Jenna;Raines, Samuel L. M.;James, Katherine S.;Sutton, Cameron;Sanchez, Anthony;Emery, Ann;Sampey, Gavin;Ferris, Robert;Allard, Brigitte;Ghofrani, Simon;Kirchherr, Jennifer L.;Baker, Caroline;Kuruc, JoAnn D.;Gay, Cynthia L.;James, Lindsey I.;Wu, Guoxin;Zuck, Paul;Rioja, Inmaculada;Furze, Rebecca C.;Prinjha, Rab K.;Howell, Bonnie J.;Swanstrom, Ronald;Browne, Edward P.;Strahl, Brian D.;Dunham, Richard M.;Archin, Nancie M.;Margolis, David M.
在抑制性抗逆转录病毒治疗（ART）期间，使用凋亡蛋白抑制剂（IAP）拮抗剂（IAPi）用于HIV治愈的潜伏逆转策略诱导了前所未有的潜伏储库表达水平，而没有免疫毒性。然而，储层的完全目标可能需要组合方法。针对IAPi组合伴侣的Jurkat潜伏期模型筛选证明了用溴结构域（BD）和末端外结构域蛋白抑制剂（BETi）的协同潜伏期逆转。使用CRISPR-CAS 9和单细胞RNA-Seq的机制研究为来自ART抑制供体的CD 4 + T细胞中的IAPi加泛BET、bD选择性BET或选择性BET同种型靶向的全面离体评价提供了信息。与T细胞活化阳性对照相比，IAPi+BETi处理导致细胞相关的HIV gag RNA的显著诱导，但完全伸长和tat-rev RNA的诱导较少。使用超灵敏p24测定，IAPi+BETi在CD 4 + T细胞的大量培养物中导致HIV蛋白诱导，但使用标准定量病毒生长测定，没有导致增强的病毒生长频率。本研究将HIV转录延长和剪接定义为潜伏逆转后潜伏HIV蛋白表达的重要障碍，描绘了BET蛋白及其BD在HIV潜伏中的作用，并为在HIV潜伏动物模型中探索IAPi+BETi提供了理论基础。
Latency reversal strategies for HIV cure using inhibitor of apoptosis protein (IAP) antagonists (IAPi) induce unprecedented levels of latent reservoir expression without immunotoxicity during suppressive antiretroviral therapy (ART). However, full targeting of the reservoir may require combinatorial approaches. A Jurkat latency model screen for IAPi combination partners demonstrated synergistic latency reversal with bromodomain (BD) and extraterminal domain protein inhibitors (BETi). Mechanistic investigations using CRISPR-CAS9 and single-cell RNA-Seq informed comprehensive ex vivo evaluations of IAPi plus pan-BET, bD-selective BET, or selective BET isoform targeting in CD4+ T cells from ART-suppressed donors. IAPi+BETi treatment resulted in striking induction of cell-associated HIV gag RNA, but lesser induction of fully elongated and tat-rev RNA compared with T cell activation–positive controls. IAPi+BETi resulted in HIV protein induction in bulk cultures of CD4+ T cells using an ultrasensitive p24 assay, but did not result in enhanced viral outgrowth frequency using a standard quantitative viral outgrowth assay. This study defines HIV transcriptional elongation and splicing as important barriers to latent HIV protein expression following latency reversal, delineates the roles of BET proteins and their BDs in HIV latency, and provides a rationale for exploration of IAPi+BETi in animal models of HIV latency.