Study of two tyrosine kinase inhibitors on growth and signal transduction in polycythemia vera
Study of two tyrosine kinase inhibitors on growth and signal transduction in polycythemia vera
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DOI：
10.1016/j.exphem.2007.08.018
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发表时间：
2007-11-01
影响因子：
2.6
通讯作者：
Prchal, Josef T.
中科院分区：
文献类型：
--
作者：
Gaikwad, Amos;Prchal, Josef T.
Objective. Janus激酶2 V617 F（JAK 2（V617 F））的激活体细胞突变存在于大多数真性红细胞增多症（PV）患者中。我们研究了两种有效的酪氨酸激酶抑制剂（TKI），AEE 788和AMN 107，在体外对携带这种突变的细胞的疗效。我们采用表达野生型JAK 2和突变型JAK 2（V617 F）的报告细胞、携带JAK 2（1617 F）的人红白血病细胞（HEL），通过细胞增殖试验、Annexin-V/碘化丙啶染色以及相关细胞信号传导和凋亡事件研究这些TKI的疗效。将这些数据与在液体培养物中生长的离体扩增的天然人红系PV祖细胞进行比较。AEE 788显示出时间和剂量依赖性生长抑制作用，其在表达JAK 2的FDCP细胞（V617 F）和HEL细胞中比在表达野生型JAK 2的细胞中更大。AEE 788引起Akt（（S243））和信号转导和转录激活因子5（（Y 694））蛋白的去磷酸化，增加Annexin-V结合和caspase-3切割，表明诱导凋亡。我们还观察到AEE 788介导的热休克蛋白70和90抗凋亡蛋白的减少。类似地，天然PV红系祖细胞显示出比正常红系祖细胞对AEE 788更敏感。AEE 788还对PV特异性红细胞集落产生剂量依赖性抑制。然而，尼洛替尼（AMN 107）缺乏特异性，需要高浓度（> 8 μ M）才能抑制携带JAK 2（V617 F）的细胞的生长。我们的数据表明，AEE 788通过下调增殖和抗凋亡调节蛋白发挥其凋亡活性。据我们所知，这是第一份证明AEE 788对PV红系祖细胞作用的报告。对PV和正常祖细胞的不同作用表明，AEE 788在治疗PV和其他JAK 2（V617 F）阳性恶性血液病中具有潜力。(C)2007 ISEH -血液学和干细胞学会。爱思唯尔公司出版
Objective. An activating somatic mutation of Janus kinase 2 V617F (JAK2(V617F)) is present in most polycythemia vera (PV) patients. We studied efficacy of two potent tyrosine kinase inhibitors (TKI), AEE788 and AMN107, in vitro on cells bearing this mutation.Materials and Methods. We employed reporter cells expressing wild-type JAK2 and mutant JAK2 (V617F), human erythroleukemic cells (HEL) carrying JAK2 (1617F) to study the efficacy of these TKIs by cell proliferation assay, Annexin-V/propidium iodide staining, and on relevant cell-signaling and apoptotic events. These data were compared to ex vivo expanded native human erythroid PV progenitor cells grown in liquid cultures.Results. AEE788 showed a time- and dose-dependent growth inhibitory effect that was greater in FDCP cells expressing JAK2(V617F) and HEL cells than in cells expressing wildtype JAK2. AEE788 caused dephosphorylation of Akt((S243)) and signal transducer and activator of transcription 5((Y694)) proteins, increase in Annexin-V binding and caspase-3 cleavage, suggesting induction of apoptosis. We also observed AEE788-mediated decrease in heat shock protein 70 and 90 antiapoptotic proteins. Similarly, native PV erythroid progenitors showed more sensitivity to AEE788 than normal erythroid progenitors. AEE788 also exerted dose-dependent inhibition of PV-specific erythroid colonies. Nilotinib (AMN107) however, lacked specificity and required high (> 8 mu M) concentrations to inhibit growth of JAK2 (V617F) -carrying cells.Conclusion. Our data suggest that AEE788 exerts its apoptotic activity via downregulation of proliferative and antiapoptotic regulatory proteins. To our knowledge, this is the first report demonstrating the effect of AEE788 on PV erythroid progenitors. Differential effects on PV and normal progenitor cells suggest AEE788 has potential in the treatment of PV and other JAK2(V617F) positive hematologic malignancies. (C) 2007 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc.