CRIF1-CDK2 Interface Inhibitors: An Unprecedented Strategy for Modulation of Cell Radiosensitivity
CRIF1-CDK2 Interface Inhibitors: An Unprecedented Strategy for Modulation of Cell Radiosensitivity
复制标题
CRIF1-CDK2 界面抑制剂：前所未有的细胞放射敏感性调节策略
DOI：
10.1021/jacs.8b10207
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发表时间：
2019-01-30
影响因子：
15
通讯作者：
Li, Zhongjun
中科院分区：
文献类型：
--
作者：
Ran, Qian;Xiang, Yang;Li, Zhongjun
细胞周期蛋白依赖性激酶（CDKs）由于其在细胞周期阶段的关键参与，是涉及致瘤事件的历史性治疗靶点。然而，选择性已被证明是一个瓶颈，导致反复失败。之前，我们报道了cr6相互作用因子1 (CRIF1)，通过与CDK2相互作用作为细胞周期负调节因子。在目前的报告中，我们通过计算机研究确定了CRIF1- cdk2相互作用界面，并通过使用40678种药物样化合物对CRIF1进行虚拟筛选，确定了入围的界面抑制剂。这些化合物通过细胞增殖实验进行了测试，发现其中4种分子选择性地抑制骨肉瘤（OS）细胞系的增殖，但不影响正常的骨间充质干细胞（BMSC）。一项结合研究显示，这些抑制剂对CRIF1具有显著的亲和力。更重要的是，用电离辐射（IR）和性能最好的抑制剂联合处理OS细胞，将IR抑制电位从19.9%显著提高到59.6%。这是通过选择性地促进OS细胞中与CDK2过度激活相关的G2/M阻滞和凋亡而发生的，而不是在BMSC中发生的，并得到了显著的CDK2磷酸化修饰的支持。通过siRNA处理敲除CRIF1显示出与界面抑制剂相似的效果。总之，我们证实了新的先导分子的鉴定，这可能为克服肿瘤细胞的选择性问题和增强肿瘤细胞的放射敏感性提供新的治疗方法，为不同癌症类型的cdk靶向开辟了一个概念上的新策略。
Cyclin-dependent kinases (CDKs) are historic therapeutic targets implicated in tumorigenic events due to their critical involvement in the cell cycle phase. However, selectivity has proven to be a bottleneck, causing repeated failures. Previously, we reported CR6-interacting factor 1 (CRIF1), acting as a cell cycle negative regulator through interaction with CDK2. In the current report, we identified the CRIF1-CDK2 interaction interface by in silico studies and shortlisted interface inhibitors through virtual screening on CRIF1 using 40 678 drug-like compounds. These compounds were tested by cell proliferation assay, and four of these molecules were found to selectively inhibit the proliferation of osteosarcoma (OS) cell lines, but do not affect normal bone mesenchymal stem cells (BMSC). A binding study reveals significant affinities of the inhibitors on CRIF1. More importantly, treatment of the OS cells with a combination of ionizing radiation (IR) and the best-performing inhibitors remarkably increased IR inhibition potential from 19.9% to 59.6%. This occurred by selectively promoting G2/M arrest and apoptosis related to CDK2 overactivation in OS cells but not in BMSC and was supported by significant CDK2 phosphorylation modifications. Knocking down of CRIF1 by siRNA treatment showed similar effects to the interface inhibitors. Together we substantiate the identification of novel lead molecules, which may provide a new treatment to overcome selectivity issues and enhance the radiosensitivity of tumor cells, opening a conceptually novel strategy of CDK-targeting for different cancer types.