Phenotype-based drug screening reveals association between venetoclax response and differentiation stage in acute myeloid leukemia
Phenotype-based drug screening reveals association between venetoclax response and differentiation stage in acute myeloid leukemia
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DOI：
10.3324/haematol.2018.214882
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发表时间：
2020-03-01
期刊：
影响因子：
10.1
通讯作者：
Heckman, Caroline A.
中科院分区：
文献类型：
--
作者：
Kuusanmaki, Heikki;Leppa, Aino-Maija;Heckman, Caroline A.
体外药物测试是一种很有前途的方法来确定急性髓性白血病（AML）的新治疗策略。然而，准确的母细胞特异性药物反应不能用均匀的“添加-混合-测量”细胞活力测定法来测量。在这项研究中，我们实施了一种基于流式细胞术的方法，同时评估了34例原发性AML样本中不同细胞群对7种药物和27种合理药物组合的体外敏感性。我们的数据表明，AML样本中存在的不同细胞群对靶向治疗具有不同的敏感性。特别是，FAB M0/1 AML的母细胞对venetoclax表现出高度敏感性。相比之下，大量存在于M4/5亚型中的分化单核细胞对Bcl-2抑制表现出抗性，而相同样品中的未成熟细胞对Bcl-2抑制敏感，这突出了blast特异性读数的重要性。因此，在总单核细胞分数中，BCL2/MCL1基因表达比在M0/1中最高，在M4/5 AML中最低。在7种测试药物中，venetoclax具有最高的blast特异性毒性，并且venetoclax与MEK抑制剂trametinib或JAK抑制剂ruxolitinib联合有效靶向所有venetoclax耐药的blast。总之，我们发现靶向药物，特别是Bcl-2抑制剂venetoclax的体外疗效受到细胞类型的影响，并且可以通过基于流式细胞术的方法来评估准确的细胞特异性药物反应。
Ex vivo drug testing is a promising approach to identify novel treatment strategies for acute myeloid leukemia (AML). However, accurate blast- specific drug responses cannot be measured with homogeneous "add-mix-measure" cell viability assays. In this study, we implemented a flow cytometry-based approach to simultaneously evaluate the ex vivo sensitivity of different cell populations in 34 primary AML samples to seven drugs and 27 rational drug combinations. Our data demonstrate that different cell populations present in AML samples have distinct sensitivity to targeted therapies. Particularly, blast cells of FAB M0/1 AML showed high sensitivity to venetoclax. In contrast, differentiated monocytic cells abundantly present in M4/5 subtypes showed resistance to Bcl-2 inhibition, whereas immature blasts in the same samples were sensitive, highlighting the importance of blast-specific readouts. Accordingly, in the total mononuclear cell fraction the highest BCL2/MCL1 gene expression ratio was observed in M0/1 and the lowest in M4/5 AML. Of the seven tested drugs, venetoclax had the highest blast-specific toxicity, and combining venetoclax with either MEK inhibitor trametinib or JAK inhibitor ruxolitinib effectively targeted all venetoclax-resistant blasts. In conclusion, we show that ex vivo efficacy of targeted agents and particularly Bcl-2 inhibitor venetoclax is influenced by the cell type, and accurate blast-specific drug responses can be assessed with a flow cytometry-based approach.