Bone marrow-derived mesenchymal stromal cells promote resistance to tyrosine kinase inhibitors in chronic myeloid leukemia via the IL-7/JAK1/STAT5 pathway
Bone marrow-derived mesenchymal stromal cells promote resistance to tyrosine kinase inhibitors in chronic myeloid leukemia via the IL-7/JAK1/STAT5 pathway
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骨髓间充质基质细胞通过 IL-7/JAK1/STAT5 途径促进慢性粒细胞白血病对酪氨酸激酶抑制剂的耐药性
DOI：
10.1074/jbc.ra119.008037
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发表时间：
2019
影响因子：
4.8
通讯作者：
Li Jian
中科院分区：
文献类型：
--
作者：
Zhang Xiaoyan;Tu Huaijun;Yang Yazhi;Jiang Xiaoyan;Hu Xianliang;Luo Qidong;Li Jian
慢性粒细胞白血病（Chronic myeloid leukemia，CML）是由RhoGEF的BCR激活蛋白（activator of RhoGEF，BCR）与非受体酪氨酸激酶（nonreceptor tyrosine kinase，ABL）原癌基因融合而引起的。尽管酪氨酸激酶抑制剂（TKI）伊马替尼（IM）和尼洛替尼（NI）在治疗CML方面具有显著疗效，但某些患者的恶性肿瘤对TKI产生耐药性。在这里，我们通过Ficoll-Hypaque密度梯度离心法从几名CML患者中分离出骨髓（BM）来源的间充质干细胞（MSC），用于与K562和BV 173细胞（含或不含TKI）共培养。我们使用实时定量PCR来评估MSC中白细胞介素7（IL-7）的表达水平，并采用免疫印迹来监测BCR/ABL、磷脂酰肌醇3-激酶（PI 3 K）/AKT和JAK/STAT信号通路中的蛋白表达。我们还使用异种移植肿瘤模型来检测不同MSC对CML细胞的体内作用。来自IM耐药CML患者的MSC保护K562和BV 173细胞免受IM或NI诱导的细胞死亡，这种保护是由于MSC分泌的IL-7增加。此外，IM耐药CML患者BM中的IL-7水平显著高于健康供体或IM敏感CML患者。IL-7通过BCR/ABL非依赖性激活JAK 1/STAT 5信号通路诱导IM和NI抵抗，而JAK 3/STAT 5和PI 3 K/AKT信号通路则不受IL-7或JAK 1基因敲除的影响，IL-7或JAK 1基因敲除可在体外和体内阻断IL-7介导的STAT 5磷酸化和IM抵抗。由于BM中的高IL-7水平通过JAK 1/STAT 5信号传导的BCR/ABL非依赖性激活介导TKI抗性，因此TKI与IL-7/JAK 1/STAT 5抑制剂的组合可能对管理CML具有显著效用。
Chronic myeloid leukemia (CML) is caused by the fusion of the BCR activator of RhoGEF and GTPase activating protein (BCR) andABLproto-oncogene, the nonreceptor tyrosine kinase (ABL) genes. Although the tyrosine kinase inhibitors (TKIs) imatinib (IM) and nilotinib (NI) have remarkable efficacy in managing CML, the malignancies in some patients become TKI-resistant. Here, we isolated bone marrow (BM)-derived mesenchymal stem cells (MSCs) from several CML patients by Ficoll-Hypaque density-gradient centrifugation for coculture with K562 and BV173 cells with or without TKIs. We used real-time quantitative PCR to assess the level of interleukin 7 (IL-7) expression in the MSCs and employed immunoblotting to monitor protein expression in the BCR/ABL, phosphatidylinositol 3-kinase (PI3K)/AKT, and JAK/STAT signaling pathways. We also used a xenograft tumor model to examine thein vivoeffect of different MSCs on CML cells. MSCs from patients with IM-resistant CML protected K562 and BV173 cells against IM- or NI-induced cell death, and this protection was due to increased IL-7 secretion from the MSCs. Moreover, IL-7 levels in the BM of patients with IM-resistant CML were significantly higher than in healthy donors or IM-sensitive CML patients. IL-7 elicited IM and NI resistance via BCR/ABL-independent activation of JAK1/STAT5 signaling, but not of JAK3/STAT5 or PI3K/AKT signaling.IL-7orJAK1gene knockdown abrogated IL-7–mediated STAT5 phosphorylation and IM resistancein vitroandin vivo. Because high IL-7 levels in the BM mediate TKI resistance via BCR/ABL-independent activation of JAK1/STAT5 signaling, combining TKIs with IL-7/JAK1/STAT5 inhibition may have significant utility for managing CML.