Combination of panobinostat with ponatinib synergistically overcomes imatinib-resistant CML cells.
Combination of panobinostat with ponatinib synergistically overcomes imatinib-resistant CML cells.
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DOI：
10.1111/cas.12965
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发表时间：
2016-07
期刊：
影响因子：
5.7
通讯作者：
Ueda T
中科院分区：
文献类型：
--
作者：
Matsuda Y;Yamauchi T;Hosono N;Uzui K;Negoro E;Morinaga K;Nishi R;Yoshida A;Kimura S;Maekawa T;Ueda T
CML伊马替尼（IM）耐药的主要机制是通过BCR‐ABL基因扩增或突变重新激活ABL激酶。我们研究了泛ABL酪氨酸激酶抑制剂泊那替尼和泛组蛋白脱乙酰酶抑制剂帕比司他对IM耐药CML细胞的体外细胞毒性。评价了两种不同的IM耐药细胞系K562/IM-R1和Ba/F3/T315 I，并与其各自的亲本细胞系K562和Ba/F3进行了比较。K562/IM-R1由于基因扩增而过表达BCR-ABL。Ba/F3/T315 I用编码T315 I突变的BCR-ABL的BCR-ABL基因转染。Ponatinib抑制K562/IM-R1和Ba/F3/T315 I的生长与其抑制其亲本细胞一样有效，IC 50为2-30 nM。帕比司他也类似地抑制所有细胞系的生长，IC 50为40-51 nM。这伴随着组蛋白去乙酰化酶活性降低，诱导组蛋白H3乙酰化，以及热休克蛋白70蛋白水平升高，这表明热休克蛋白90对BCR-ABL及其降解的伴侣蛋白功能被破坏。重要的是，在所有细胞系中，泊那替尼与帕比司他的组合显示出协同生长抑制作用，并且诱导的细胞凋亡水平高于每种药物单独诱导的细胞凋亡的总和。Ponatinib不仅抑制BCR-ABL的磷酸化，还抑制K562/IM-R1和Ba/F3/T315 I中下游信号转导子和转录激活因子5、蛋白激酶B和ERK 1/2的磷酸化，在Ponatinib中加入帕比司他进一步抑制这些磷酸化。总之，帕比司他增强了泊那替尼对IM耐药CML细胞（包括T315 I突变的BCR-ABL细胞）的细胞毒性。
The major mechanism of imatinib (IM) resistance of CML is the reactivation of ABL kinase either through BCR‐ABL gene amplification or mutation. We investigated the cytotoxicity of a pan‐ABL tyrosine kinase inhibitor, ponatinib, and a pan‐histone deacetylase inhibitor, panobinostat, against IM‐resistant CML cells in vitro. Two different IM‐resistant cell lines, K562/IM‐R1 and Ba/F3/T315I were evaluated in comparison with their respective, parental cell lines, K562 and Ba/F3. K562/IM‐R1 overexpressed BCR‐ABL due to gene amplification. Ba/F3/T315I was transfected with a BCR‐ABL gene encoding T315I‐mutated BCR‐ABL. Ponatinib inhibited the growth of both K562/IM‐R1 and Ba/F3/T315I as potently as it inhibited their parental cells with an IC 50 of 2–30 nM. Panobinostat also similarly inhibited the growth of all of the cell lines with an IC 50 of 40–51 nM. This was accompanied by reduced histone deacetylase activity, induced histone H3 acetylation, and an increased protein level of heat shock protein 70, which suggested disruption of heat shock protein 90 chaperone function for BCR‐ABL and its degradation. Importantly, the combination of ponatinib with panobinostat showed synergistic growth inhibition and induced a higher level of apoptosis than the sum of the apoptosis induced by each agent alone in all of the cell lines. Ponatinib inhibited phosphorylation not only of BCR‐ABL but also of downstream signal transducer and activator of transcription 5, protein kinase B, and ERK1/2 in both K562/IM‐R1 and Ba/F3/T315I, and the addition of panobinostat to ponatinib further inhibited these phosphorylations. In conclusion, panobinostat enhanced the cytotoxicity of ponatinib towards IM‐resistant CML cells including those with T315I‐mutated BCR‐ABL.