Enhanced dependency of KRAS-mutant colorectal cancer cells on RAD51-dependent homologous recombination repair identified from genetic interactions in Saccharomyces cerevisiae.
Enhanced dependency of KRAS-mutant colorectal cancer cells on RAD51-dependent homologous recombination repair identified from genetic interactions in Saccharomyces cerevisiae.
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DOI：
10.1002/1878-0261.12040
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发表时间：
2017-05
影响因子：
6.6
通讯作者：
Khanna KK
中科院分区：
文献类型：
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作者：
Kalimutho M;Bain AL;Mukherjee B;Nag P;Nanayakkara DM;Harten SK;Harris JL;Subramanian GN;Sinha D;Shirasawa S;Srihari S;Burma S;Khanna KK
激活KRAS突变驱动结直肠癌肿瘤发生并影响抗EGFR靶向治疗的反应。尽管最近在理解Ras信号传导生物学和使用BRAF抑制剂治疗黑色素瘤方面取得了进展，但没有靶向药物对KRAS突变型癌症有效，主要是由于代偿途径的激活。在这里，通过利用酵母中最大的合成致死遗传相互作用组，我们确定KRAS突变的结直肠癌细胞增强了同源重组修复（HRR）信号传导。我们发现KRAS突变导致复制叉的减慢和停滞以及DNA损伤的积累。此外，我们发现，与基因互补的同基因细胞相比，KRAS突变型HCT 116细胞的MYC介导的RAD 51表达增加，辐射诱导的DNA双链断裂（DSB）的RAD 51募集相应增加。使用RNA干扰去除MYC显著降低IR诱导的RAD 51灶形成和HRR。相反，HA标记的野生型（WT）MYC或磷酸化突变体S62 A的过表达增加了RAD 51蛋白水平，从而增加了IR诱导的RAD 51病灶。同样，RAD 51的消耗通过增加DSB选择性地诱导HCT 116突变细胞的凋亡。靶向HRR信号传导的药理学抑制与PARP抑制选择性结合杀死KRAS突变细胞。有趣的是，这些差异在KRAS WT和突变细胞（DLD-1）的第二个同基因对中没有观察到，可能是由于它们的生存不依赖于KRAS突变。因此，我们的数据突出了KRAS突变依赖性细胞通过上调MYC-RAD 51表达在体外驱动HRR的可能机制。这些数据可能提供了一个有前途的治疗漏洞，在结直肠癌细胞窝藏否则nondruggable KRAS突变，这值得进一步研究在体内。
Activating KRAS mutations drive colorectal cancer tumorigenesis and influence response to anti‐EGFR‐targeted therapy. Despite recent advances in understanding Ras signaling biology and the revolution in therapies for melanoma using BRAF inhibitors, no targeted agents have been effective in KRAS‐mutant cancers, mainly due to activation of compensatory pathways. Here, by leveraging the largest synthetic lethal genetic interactome in yeast, we identify that KRAS‐mutated colorectal cancer cells have augmented homologous recombination repair (HRR) signaling. We found that KRAS mutation resulted in slowing and stalling of the replication fork and accumulation of DNA damage. Moreover, we found that KRAS‐mutant HCT116 cells have an increase in MYC‐mediated RAD51 expression with a corresponding increase in RAD51 recruitment to irradiation‐induced DNA double‐strand breaks (DSBs) compared to genetically complemented isogenic cells. MYC depletion using RNA interference significantly reduced IR‐induced RAD51 foci formation and HRR. On the contrary, overexpression of either HA‐tagged wild‐type (WT) MYC or phospho‐mutant S62A increased RAD51 protein levels and hence IR‐induced RAD51 foci. Likewise, depletion of RAD51 selectively induced apoptosis in HCT116‐mutant cells by increasing DSBs. Pharmacological inhibition targeting HRR signaling combined with PARP inhibition selectivity killed KRAS‐mutant cells. Interestingly, these differences were not seen in a second isogenic pair of KRAS WT and mutant cells (DLD‐1), likely due to their nondependency on the KRAS mutation for survival. Our data thus highlight a possible mechanism by which KRAS‐mutant‐dependent cells drive HRR in vitro by upregulating MYC‐RAD51 expression. These data may offer a promising therapeutic vulnerability in colorectal cancer cells harboring otherwise nondruggable KRAS mutations, which warrants further investigation in vivo.
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影响因子：
64.8
作者：
Bartkova, Jirina;Rezaei, Nousin;Gorgoulis, Vassilis G.
通讯作者：
Gorgoulis, Vassilis G.
影响因子：
9
作者：
通讯作者：
--
DOI：
10.1158/1078-0432.ccr-13-1607
发表时间：
2014-03-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Gil del Alcazar CR;Hardebeck MC;Mukherjee B;Tomimatsu N;Gao X;Yan J;Xie XJ;Bachoo R;Li L;Habib AA;Burma S
通讯作者：
Burma S
影响因子：
5.7
作者：
Davies, Barry R.;Logie, Armelle;Smith, Paul D.
通讯作者：
Smith, Paul D.
影响因子：
3.8
作者：
Chen Y;Xu J;Borowicz S;Collins C;Huo D;Olopade OI
通讯作者：
Olopade OI