Department of Hematology, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing, Jiangsu Province, PR China.
Key Laboratory of Hematology of Nanjing Medical University, Jiangsu Province Hospital, Nanjing, 210029, Jiangsu Province, PR China.
Oncogene. 2018 May;37(21):2837-2849. doi: 10.1038/s41388-018-0157-8. Epub 2018 Mar 7.
Resistance to the BCR-ABL tyrosine kinase inhibitor (TKI) remains a challenge for curing the disease in chronic myeloid leukemia (CML) patients as leukemia cells may survive through BCR-ABL kinase activity-independent signal pathways. To gain insight into BCR-ABL kinase activity-independent mechanisms, we performed an initial bioinformatics screen and followed by a quantitative PCR screen of genes that were elevated in CML samples. A total of 33 candidate genes were identified to be highly expressed in TKIs resistant patients. Among those genes, 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3), controlling the limiting step of glycolysis, was found to be strongly associated with TKIs resistance. PFKFB3 knockdown or pharmacological inhibition of its kinase activity markedly enhanced the sensitivity of CML cells to TKIs. Furthermore, pharmacological inhibition of PFKFB3 inhibited CML cells growth and significantly prolonged the survival of both allograft and xenograft CML mice. ChIP-seq data analysis combined with subsequent knockdown experiment showed that the Ets transcription factor PU.1 regulated the elevated expression of PFKFB3 in TKIs-resistant CML cells. Therefore, our results showed that targeting PFKFB3 sensitizes CML cells to TKIs and PFKFB3 may be a potential BCR-ABL kinase activity-independent mechanism in CML.
BCR-ABL 酪氨酸激酶抑制剂 (TKI) 的耐药性仍然是治愈慢性髓性白血病 (CML) 患者疾病的一个挑战,因为白血病细胞可能通过 BCR-ABL 激酶活性非依赖性信号通路存活。为了深入了解 BCR-ABL 激酶活性非依赖性机制,我们进行了初始的生物信息学筛选,随后对 CML 样本中升高的基因进行了定量 PCR 筛选。总共鉴定出 33 个候选基因在 TKI 耐药患者中高度表达。在这些基因中,控制糖酵解限速步骤的 6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶 3 (PFKFB3) 被发现与 TKI 耐药性强烈相关。PFKFB3 的敲低或激酶活性的药理学抑制显著增强了 CML 细胞对 TKI 的敏感性。此外,PFKFB3 的药理学抑制抑制了 CML 细胞的生长,并显著延长了同种异体和异种移植 CML 小鼠的存活时间。ChIP-seq 数据分析结合随后的敲低实验表明,Ets 转录因子 PU.1 调节 TKI 耐药 CML 细胞中 PFKFB3 的上调表达。因此,我们的结果表明,靶向 PFKFB3 可使 CML 细胞对 TKI 敏感,PFKFB3 可能是 CML 中一种潜在的 BCR-ABL 激酶活性非依赖性机制。