Combined Inhibition of Rad51 and Wee1 Enhances Cell Killing in HNSCC Through Induction of Apoptosis Associated With Excessive DNA Damage and Replication Stress.
Combined Inhibition of Rad51 and Wee1 Enhances Cell Killing in HNSCC Through Induction of Apoptosis Associated With Excessive DNA Damage and Replication Stress.
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DOI：
10.1158/1535-7163.mct-20-0252
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发表时间：
2021-07
影响因子：
5.7
通讯作者：
Osman AA
中科院分区：
文献类型：
--
作者：
Lindemann A;Patel AA;Tang L;Tanaka N;Gleber-Netto FO;Bartels MD;Wang L;McGrail DJ;Lin SY;Frank SJ;Frederick MJ;Myers JN;Osman AA
尽管在手术、化疗和放射治疗方面取得了进展，但晚期头颈部鳞状细胞癌(HNSCC)的治疗选择有限，生存率仍然很低。因此，迫切需要有效的治疗方法。最近，选择性地利用DNA损伤和复制应激反应已成为治疗癌症的一种新方法。Wee1激酶和RAD51重组酶是参与调节癌细胞复制应激和同源重组修复的两种蛋白质。在本研究中，我们研究了RAD51抑制剂(B02)和Wee1抑制剂(AZD1775)在体内外对不同HNSCC细胞株的联合作用。克隆存活实验表明，B02与AZD1775在体外对所有受试HNSCC细胞株均有协同作用。这些药物之间的协同作用与强迫激活CDK1和降低Chk1磷酸化有关，导致过度的DNA损伤和复制应激，最终导致异常有丝分裂和细胞凋亡。我们的结果表明，在HPV阳性的HNSCC患者中，RAD51mRNA的高表达与较差的生存相关。与HPV阴性HNSCC相比，B02和AZD1775的联合使用显著抑制了HPV阳性HNSCC小鼠体内的肿瘤生长。这种不同的敏感性似乎与HPV阳性肿瘤由于E6和E7癌基因的转化而具有更多的体内复制压力有关。此外，B02的加入在体内外对HPV阴性HNSCC肿瘤具有放射增敏作用。总之，我们的数据暗示，RAD51和Wee1抑制剂的合理组合有望成为合成致死疗法，特别是在高危HPV阳性的HNSCC中。
Despite advances in surgery, chemotherapy, and radiation, there are limited treatment options for advanced head and neck squamous cell carcinoma (HNSCC) and survival remains very poor. Therefore, effective therapies are desperately needed. Recently, selective exploitation of DNA damage and replication stress responses has become a novel approach for cancer treatment. Wee1 kinase and Rad51 recombinase are two proteins involved in regulating replication stress and homologous recombination repair in cancer cells. In this study, we investigated the combined effect of Rad51 inhibitor (B02) and Wee1 inhibitor (AZD1775) in vitro and in vivo in various HNSCC cell lines. Clonogenic survival assays demonstrated that B02 synergized with AZD1775 in vitro in all HNSCC cell lines tested. The synergy between these drugs was associated with forced CDK1 activation and reduced Chk1 phosphorylation leading to induction of excessive DNA damage and replication stress, culminating in aberrant mitosis and apoptosis. Our results showed that elevated Rad51 mRNA expression correlated with worse survival in HNSCC patients with HPV-positive tumors. The combination of B02 and AZD1775 significantly inhibited tumor growth in vivo in mice bearing HPV-positive HNSCC tumors as compared to HPV-negative HNSCC. This differential sensitivity appears to be linked to HPV-positive tumors having more in vivo endogenous replication stress owing to transformation by E6 and E7 oncogenes. Furthermore, addition of B02 radiosensitized the HPV-negative HNSCC tumors in vitro and in vivo. In conclusion, our data implicate that a novel rational combination with Rad51 and Wee1 inhibitors holds promise as synthetic lethal therapy, particularly in high-risk HPV-positive HNSCC.