Mechanism by Which PF-3758309, a Pan Isoform Inhibitor of p21-Activated Kinases, Blocks Reactivation of HIV-1 Latency.
Mechanism by Which PF-3758309, a Pan Isoform Inhibitor of p21-Activated Kinases, Blocks Reactivation of HIV-1 Latency.
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PF-3758309（p21 激活激酶的泛亚型抑制剂）阻止 HIV-1 潜伏期重新激活的机制。
DOI：
10.3390/biom13010100
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发表时间：
2023-01-04
期刊：
影响因子：
5.5
通讯作者：
中科院分区：
文献类型：
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作者：
“封锁和锁定”战略是一种可能导致对HIV-1感染进行消毒治疗的方法。“阻断”指的是化合物抑制潜伏的HIV-1前病毒转录的能力，而“锁定”指的是它诱导永久前病毒沉默的能力。我们先前发现PF-3758309，一种p21激活的蛋白激酶(PAK)的泛异构体抑制物，是HIV-1潜伏期逆转的有效抑制物。本研究的目的是明确S所涉及的机制。我们发现，24ST1NLESG细胞(HIV-1潜伏的细胞系模型)和纯化的CD4+幼稚T细胞和中央记忆T细胞都表达高水平的PAK2，而低水平的PAK1和PAK4。在24ST1NLESG细胞中敲除PAK1或PAK2，而不是PAK4，导致HIV-1潜伏期逆转的幅度略有下降，但在统计学上具有显著意义。PAK1的过表达显著增加了潜伏期逆转的幅度。磷酸化蛋白质阵列分析显示，PF-3758309下调了NF-κB信号通路，这是PF-3758309抑制潜伏期逆转的最可能机制。最后，我们使用细胞热位移分析结合高效液相色谱和质谱仪来确定PF-3758309的非靶点结合是否对其活性有贡献。在24ST1NLESG细胞和外周血单核细胞中，PF-3758309与丝裂原激活的蛋白激酶1和蛋白激酶A结合；然而，这两种激酶中的任何一种都不影响HIV-1潜伏期的逆转。综上所述，我们的研究表明，PAK1和PAK2在维持HIV-1潜伏期方面发挥了关键作用。
The “block and lock” strategy is one approach that might elicit a sterilizing cure for HIV-1 infection. The “block” refers to a compound’s ability to inhibit latent HIV-1 proviral transcription, while the “lock” refers to its capacity to induce permanent proviral silencing. We previously identified PF-3758309, a pan-isoform inhibitor of p21-activated kinases (PAKs), as a potent inhibitor of HIV-1 latency reversal. The goal of this study was to define the mechanism(s) involved. We found that both 24ST1NLESG cells (a cell line model of HIV-1 latency) and purified CD4+ naïve and central memory T cells express high levels of PAK2 and lower levels of PAK1 and PAK4. Knockdown of PAK1 or PAK2, but not PAK4, in 24ST1NLESG cells resulted in a modest, but statistically significant, decrease in the magnitude of HIV-1 latency reversal. Overexpression of PAK1 significantly increased the magnitude of latency reversal. A phospho-protein array analysis revealed that PF-3758309 down-regulates the NF-κB signaling pathway, which provides the most likely mechanism by which PF-3758309 inhibits latency reversal. Finally, we used cellular thermal shift assays combined with liquid chromatography and mass spectrometry to ascertain whether PF-3758309 off-target binding contributed to its activity. In 24ST1NLESG cells and in peripheral blood mononuclear cells, PF-3758309 bound to mitogen-activated protein kinase 1 and protein kinase A; however, knockdown of either of these kinases did not impact HIV-1 latency reversal. Collectively, our study suggests that PAK1 and PAK2 play a key role in the maintenance of HIV-1 latency.
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影响因子：
3.7
作者：
Mbonye, Uri;Karn, Jonathan
通讯作者：
Karn, Jonathan
PAK 激酶的结构、生物化学和生物学
影响因子：
3.5
作者：
Kumar R;Sanawar R;Li X;Li F
通讯作者：
Li F
DOI：
10.3390/pathogens11060611
发表时间：
2022-05-24
期刊：
Pathogens (Basel, Switzerland)
影响因子：
--
作者：
通讯作者：
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影响因子：
4.1
作者：
Chan JK;Greene WC
通讯作者：
Greene WC
影响因子：
4.6
作者：
Shi H;Berger EA
通讯作者：
Berger EA