Shanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine at Shanghai, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Biomolecules. 2021 May 22;11(6):781. doi: 10.3390/biom11060781.
mutations are frequently identified in acute myeloid leukemia (AML) and indicate poor prognosis. Previously, we found that the hotspot mutation DNMT3A R882H could upregulate CDK1 and induce AML in conditional knock-in mice. However, the mechanism by which CDK1 is involved in leukemogenesis of DNMT3A mutation-related AML, and whether CDK1 could be a therapeutic target, remains unclear. In this study, using fluorescence resonance energy transfer and immunoprecipitation analysis, we discovered that increased CDK1 could compete with EZH2 to bind to the PHD-like motif of DNMT3A, which may disturb the protein interaction between EZH2 and DNMT3A. Knockdown of CDK1 in OCI-AML3 cells with mutation markedly inhibited proliferation and induced apoptosis. CDK1 selective inhibitor CGP74514A (CGP) and the pan-CDK inhibitor flavopiridol (FLA) arrested OCI-AML3 cells in the G2/M phase, and induced cell apoptosis. CGP significantly increased CD163-positive cells. Moreover, the combined application of CDK1 inhibitor and traditional chemotherapy drugs synergistically inhibited proliferation and induced apoptosis of OCI-AML3 cells. In conclusion, this study highlights CDK1 overexpression as a pathogenic factor and a potential therapeutic target for DNMT3A mutation-related AML.
突变经常在急性髓系白血病 (AML) 中被发现，提示预后不良。之前，我们发现热点突变 DNMT3A R882H 可以上调 CDK1 并在条件敲入小鼠中诱导 AML。然而，CDK1 如何参与 DNMT3A 突变相关 AML 的白血病发生，以及 CDK1 是否可以作为治疗靶点，尚不清楚。在这项研究中，我们使用荧光共振能量转移和免疫沉淀分析发现，增加的 CDK1 可以与 EZH2 竞争结合到 DNMT3A 的 PHD 样结构域，这可能会干扰 EZH2 和 DNMT3A 之间的蛋白质相互作用。在携带突变的 OCI-AML3 细胞中敲低 CDK1 可显著抑制增殖并诱导细胞凋亡。CDK1 选择性抑制剂 CGP74514A (CGP) 和泛-CDK 抑制剂 flavopiridol (FLA) 将 OCI-AML3 细胞阻滞在 G2/M 期，并诱导细胞凋亡。CGP 显著增加了 CD163 阳性细胞。此外，CDK1 抑制剂与传统化疗药物联合应用可协同抑制 OCI-AML3 细胞的增殖并诱导细胞凋亡。总之，本研究强调 CDK1 过表达是 DNMT3A 突变相关 AML 的发病因素和潜在治疗靶点。
