A novel fusion of RBM6 to CSF1R in acute megakaryoblastic leukemia
A novel fusion of RBM6 to CSF1R in acute megakaryoblastic leukemia
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DOI：
10.1182/blood-2006-10-052282
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发表时间：
2007-07-01
期刊：
影响因子：
20.3
通讯作者：
Polakiewicz, Roberto D.
中科院分区：
文献类型：
--
作者：
Gu, Ting-lei;Mercher, Thomas;Polakiewicz, Roberto D.
活化的酪氨酸激酶经常涉及癌症的发病机制，包括急性髓性白血病（AML），并且是用小分子激酶抑制剂进行治疗干预的经验证的靶标。为了鉴定AML中的新型活化酪氨酸激酶，我们使用了由免疫亲和分析与质谱联用组成的发现平台，该平台鉴定了大量酪氨酸磷酸化蛋白，包括活性激酶。该方法揭示了急性巨核细胞白血病（AMKL）细胞系MKPL-1中存在活化的集落刺激因子1受体（CSF 1 R）激酶。使用siRNA和小分子抑制剂的进一步研究表明，CSF 1 R对MKPL-1细胞的生长和存活至关重要。通过对CSF 1 R编码序列的5 'RACE产生的cDNA进行DNA序列分析，鉴定了RNA结合基序6（RBM 6）基因与CSF 1 R基因的新型融合体，推测是由t（3;5）（p21;q33）易位产生的。RBM 6-CSFlR融合蛋白的表达赋予BaF 3细胞中白细胞介素-3（IL-3）非依赖性生长，并在鼠移植模型中诱导具有巨核细胞白血病特征的骨髓增殖性疾病（MPD）。这些发现确定了一个新的潜在的治疗靶点在白血病，并证明了实用程序的磷酸蛋白质策略发现酪氨酸激酶等位基因。
Activated tyrosine kinases have been frequently implicated in the pathogenesis of cancer, including acute myeloid leukemia (AML), and are validated targets for therapeutic intervention with small-molecule kinase inhibitors. To identify novel activated tyrosine kinases in AML, we used a discovery platform consisting of immunoaffinity profiling coupled to mass spectrometry that identifies large numbers of tyrosine-phosphorylated proteins, including active kinases. This method revealed the presence of an activated colony-stimulating factor 1 receptor (CSF1R) kinase in the acute megakaryoblastic leukemia (AMKL) cell line MKPL-1. Further studies using siRNA and a small-molecule inhibitor showed that CSF1R is essential for the growth and survival of MKPL-1 cells. DNA sequence analysis of cDNA generated by 5'RACE from CSF1R coding sequences identified a novel fusion of the RNA binding motif 6 (RBM6) gene to CSF1R gene generated presumably by a t(3;5)(p21;q33) translocation. Expression of the RBM6-CSFIR fusion protein conferred interleukin-3 (IL-3)-independent growth in BaF3 cells, and induces a myeloid proliferative disease (MPD) with features of megakaryoblastic leukemia in a murine transplant model. These findings identify a novel potential therapeutic target in leukemogenesis, and demonstrate the utility of phosphoproteornic strategies for discovery of tyrosine kinase alleles.