Jiang Zhiwu, Wu Di, Ye Wei, Weng Jianyu, Lai Peilong, Shi Pengcheng, Guo Xutao, Huang Guohua, Deng Qiuhua, Tang Yanlai, Zhao Hongyu, Cui Shuzhong, Lin Simiao, Wang Suna, Li Baiheng, Wu Qiting, Li Yangqiu, Liu Pentao, Pei Duanqing, Du Xin, Yao Yao, Li Peng
Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
Oncotarget. 2017 Nov 15;8(63):106382-106392. doi: 10.18632/oncotarget.22466. eCollection 2017 Dec 5.
Functional screening for compounds represents a major hurdle in the development of rational therapeutics for B-acute lymphoblastic leukemia (B-ALL). In addition, using cell lines as valid models for evaluating responses to novel drug therapies raises serious concerns, as cell lines are prone to genotypic/phenotypic drift and loss of heterogeneity . Here, we reported that OP9 cells, not OP9-derived adipocytes (OP9TA), support the growth of primary B-ALL cells . To identify the factors from OP9 cells that support the growth of primary B-ALL cells, we performed RNA-Seq to analyze the gene expression profiles of OP9 and OP9TA cells. We thus developed a defined, serum/feeder-free condition (FI76V) that can support the expansion of a range of clinically distinct primary B-ALL cells that still maintain their leukemia-initiating ability. We demonstrated the suitability of high-throughput drug screening based on our B-ALL cultured conditions. Upon screening 378 kinase inhibitors, we identified a cluster of 17 kinase inhibitors that can efficiently kill B-ALL cells . Importantly, we demonstrated the synergistic cytotoxicity of dinaciclib/BTG226 to B-ALL cells. Taken together, we developed a defined condition for the expansion of primary B-ALL cells that is suitable for high-throughput screening of novel compounds.
化合物的功能筛选是B淋巴细胞白血病（B-ALL）合理治疗药物开发中的一个主要障碍。此外，将细胞系用作评估新型药物疗法反应的有效模型引发了严重担忧，因为细胞系容易发生基因型/表型漂移和异质性丧失。在此，我们报告OP9细胞而非OP9衍生的脂肪细胞（OP9TA）支持原发性B-ALL细胞的生长。为了鉴定OP9细胞中支持原发性B-ALL细胞生长的因子，我们进行了RNA测序以分析OP9和OP9TA细胞的基因表达谱。因此，我们开发了一种明确的、无血清/无饲养层的条件（FI76V），该条件可以支持一系列临床上不同的原发性B-ALL细胞的扩增，这些细胞仍保持其白血病起始能力。我们证明了基于我们的B-ALL培养条件进行高通量药物筛选的适用性。在筛选378种激酶抑制剂后，我们鉴定出一组17种激酶抑制剂，它们可以有效杀死B-ALL细胞。重要的是，我们证明了dinaciclib/BTG226对B-ALL细胞的协同细胞毒性。综上所述，我们开发了一种用于原发性B-ALL细胞扩增的明确条件，该条件适用于新型化合物的高通量筛选。
