Discovery and development of novel DNA-PK inhibitors by targeting the unique Ku-DNA interaction.
Discovery and development of novel DNA-PK inhibitors by targeting the unique Ku-DNA interaction.
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DOI：
10.1093/nar/gkaa934
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发表时间：
2020-11-18
影响因子：
14.9
通讯作者：
Turchi JJ
中科院分区：
文献类型：
--
作者：
Gavande NS;VanderVere-Carozza PS;Pawelczak KS;Mendoza-Munoz P;Vernon TL;Hanakahi LA;Summerlin M;Dynlacht JR;Farmer AH;Sears CR;Nasrallah NA;Garrett J;Turchi JJ
DNA依赖性蛋白激酶（DNA-PK）在非同源末端连接（NHEJ）修复途径和DNA损伤反应（DDR）中起着关键作用。因此，DNA-PK已被用于开发与电离辐射（IR）组合的抗癌治疗剂。我们报告发现了一类新的DNA-PK抑制剂，其通过一种新的作用机制起作用，抑制Ku-DNA相互作用。我们已经开发了一系列高效和特异性的Ku-DNA结合抑制剂（Ku-DBi），其阻断Ku-DNA相互作用并抑制DNA-PK激酶活性。Ku-DBi直接与Ku相互作用并抑制体外NHEJ、细胞NHEJ，并增强拟辐射剂和IR的细胞活性。Ku-无效细胞的分析表明，Ku-DBi的细胞活性是Ku抑制的直接结果，因为Ku-无效细胞对Ku-DBi不敏感。在CRISPR基因编辑模型中也揭示了Ku-DBi的效用，其中我们证明了在用Ku-DBi预处理的细胞中基因插入事件的效率增加，这与抑制NHEJ和激活同源重组以促进基因插入一致。这些数据证明了通过独特的作用机制调节DNA修复途径的新系列化合物的发现和应用。
DNA-dependent protein kinase (DNA-PK) plays a critical role in the non-homologous end joining (NHEJ) repair pathway and the DNA damage response (DDR). DNA-PK has therefore been pursued for the development of anti-cancer therapeutics in combination with ionizing radiation (IR). We report the discovery of a new class of DNA-PK inhibitors that act via a novel mechanism of action, inhibition of the Ku–DNA interaction. We have developed a series of highly potent and specific Ku–DNA binding inhibitors (Ku-DBi’s) that block the Ku–DNA interaction and inhibit DNA-PK kinase activity. Ku-DBi’s directly interact with the Ku and inhibit in vitro NHEJ, cellular NHEJ, and potentiate the cellular activity of radiomimetic agents and IR. Analysis of Ku-null cells demonstrates that Ku-DBi’s cellular activity is a direct result of Ku inhibition, as Ku-null cells are insensitive to Ku-DBi’s. The utility of Ku-DBi’s was also revealed in a CRISPR gene-editing model where we demonstrate that the efficiency of gene insertion events was increased in cells pre-treated with Ku-DBi’s, consistent with inhibition of NHEJ and activation of homologous recombination to facilitate gene insertion. These data demonstrate the discovery and application of new series of compounds that modulate DNA repair pathways via a unique mechanism of action.
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DOI：
10.1073/pnas.94.25.13588
发表时间：
1997-12-09
影响因子：
11.1
作者：
Nussenzweig, A;Sokol, K;Li, GC
通讯作者：
Li, GC
影响因子：
3.8
作者：
Bachu, Ravichandra;Bergareche, Inigo;Chasin, Lawrence A.
通讯作者：
Chasin, Lawrence A.
影响因子：
5.6
作者：
Davidson D;Amrein L;Panasci L;Aloyz R
通讯作者：
Aloyz R
影响因子：
16.6
作者：
Lieber MR
通讯作者：
Lieber MR
影响因子：
7.3
作者：
Friesner, Richard A.;Murphy, Robert B.;Mainz, Daniel T.
通讯作者：
Mainz, Daniel T.