Quantitative phosphotyrosine profiling of patient-derived xenografts identifies therapeutic targets in pediatric leukemia.
Quantitative phosphotyrosine profiling of patient-derived xenografts identifies therapeutic targets in pediatric leukemia.
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DOI：
10.1158/0008-5472.can-15-2786
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发表时间：
2016-05
期刊：
影响因子：
11.2
通讯作者：
Lock RB
中科院分区：
文献类型：
--
作者：
Dolai S;Sia KC;Robbins AK;Zhong L;Heatley SL;Vincent TL;Hochgräfe F;Sutton R;Kurmasheva RT;Revesz T;White DL;Houghton PJ;Smith MA;Teachey DT;Daly RJ;Raftery MJ;Lock RB
酪氨酸激酶（TK）的激活突变会导致儿科高危急性淋巴细胞白血病（ALL）并赋予对标准化疗的耐药性。因此，迫切需要描述ALL患者中失调的TK信号传导轴，并确定可操作的激酶靶点，以制定治疗策略。在这里，我们提出了第一项研究，定量分析TK活性的异种移植患者活检的高风险儿科急性淋巴细胞白血病。我们整合了一个定量磷酸酪氨酸分析方法与“穗”稳定同位素标记的氨基酸在细胞培养（SILAC）和定量1394 I类磷酸化位点在16 ALL异种移植。此外，磷酸酪氨酸位点的分层聚类可以准确地将这些白血病分为B细胞或T细胞谱系，其中高危早期T细胞前体（ETP）和Ph样ALL聚类为一个独特的组。此外，我们通过使用特异性激酶途径抑制剂干扰四个异种移植患者样本中的ABL1、FLT3和JAK TK信号传导来验证这种方法。通过定量评估ALL异种移植模型中活化激酶的酪氨酸磷酸化状态，我们能够识别和验证临床相关靶点。因此，本研究强调了磷酸酪氨酸分析在白血病中识别临床相关激酶靶点的应用和潜力。
Activating mutations in tyrosine kinases (TK) drive pediatric high-risk acute lymphoblastic leukemia (ALL) and confer resistance to standard chemotherapy. Therefore, there is urgent need to characterize dysregulated TK signaling axes in patients with ALL and identify actionable kinase targets for the development of therapeutic strategies. Here, we present the first study to quantitatively profile TK activity in xenografted patient biopsies of high-risk pediatric ALL. We integrated a quantitative phosphotyrosine profiling method with “spike-in” stable isotope labeling with amino acids in cell culture (SILAC) and quantified 1394 class I phosphorylation sites in 16 ALL xenografts. Moreover, hierarchical clustering of phosphotyrosine sites could accurately classify these leukemias into either B- or T-cell lineages with the high-risk early T-cell precursor (ETP) and Ph-like ALL clustering as a distinct group. Furthermore, we validated this approach by using specific kinase pathway inhibitors to perturb ABL1, FLT3, and JAK TK signaling in four xenografted patient samples. By quantitatively assessing the tyrosine phosphorylation status of activated kinases in xenograft models of ALL, we were able to identify and validate clinically relevant targets. Therefore, this study highlights the application and potential of phosphotyrosine profiling for identifying clinically relevant kinase targets in leukemia.