Inhibition of DOT1L and PRMT5 promote synergistic anti-tumor activity in a human MLL leukemia model induced by CRISPR/Cas9
Inhibition of DOT1L and PRMT5 promote synergistic anti-tumor activity in a human MLL leukemia model induced by CRISPR/Cas9
复制标题
DOI：
10.1038/s41388-019-0937-9
复制
发表时间：
2019-11-14
期刊：
影响因子：
8
通讯作者：
Schneidawind, Corina
中科院分区：
文献类型：
--
作者：
Secker, Kathy-Ann;Keppeler, Hildegard;Schneidawind, Corina
MLL 重排在白血病发生中起着至关重要的作用，并且预后不良。因此，迫切需要新的治疗策略。我们使用 CRISPR/Cas9 系统生成了一种创新的白血病模型，该模型基于 100% 纯的 MLL-AF4 或 -AF9 重排细胞，这些细胞源自脐带血，在细胞培养系统中无限期生长。我们的模型具有患者细胞的表型、形态和分子特征，忠实地模拟了疾病的本质。因此，它是药理学研究的基础：抑制端粒沉默组蛋白甲基转移酶干扰物 1 样 (DOT1L) 是目前在临床试验中测试的一种特异性治疗方法。然而，由于反应有限，需要对药物组合进行进一步研究，因此成功受到限制。最近，研究表明，抑制蛋白精氨酸甲基转移酶 5 (PRMT5) 对人类细胞系和 MLL 小鼠模型具有抗肿瘤活性。在这里，我们在人类 MLL 重排模型中使用了 DOT1L 和 PRMT5 抑制剂，结果显示剂量依赖性增殖减少、细胞周期受损、分化增加、细胞凋亡、靶基因下调和化疗敏感性。引人注目的是，两种化合物的组合产生了协同抗肿瘤作用。我们的研究为新型靶向联合疗法改善 MLL 重排白血病的预后提供了强有力的理论依据。
MLL rearrangements play a crucial role in leukemogenesis and comprise a poor prognosis. Therefore, new treatment strategies are urgently needed. We used the CRISPR/Cas9 system to generate an innovative leukemia model based on 100% pure MLL-AF4 or -AF9 rearranged cells derived from umbilical cord blood with indefinite growth in cell culture systems. Our model shared phenotypical, morphological and molecular features of patient cells faithfully mimicking the nature of the disease. Thus, it serves as a fundamental basis for pharmacological studies: inhibition of histone methyltransferase disruptor of telomeric silencing 1-like (DOT1L) is one specific therapeutic approach currently tested in clinical trials. However, success was limited by restricted response warranting further investigation of drug combinations. Recently, it has been shown that the inhibition of protein arginine methyltransferase 5 (PRMT5) exhibits anti-tumoral activity against human cell lines and in MLL mouse models. Here, we used DOT1L and PRMT5 inhibitors in our human MLL-rearranged model demonstrating dose-dependent reduced proliferation, impairment of cell cycle, increasing differentiation, apoptosis, downregulation of target genes and sensitization to chemotherapy. Strikingly, the combination of both compounds led to synergistic anti-tumoral effects. Our study provides a strong rationale for novel targeted combination therapies to improve the outcome of MLL-rearranged leukemias.