Granulocyte-macrophage progenitors as candidate leukemic stem cells in blast-crisis CML
Granulocyte-macrophage progenitors as candidate leukemic stem cells in blast-crisis CML
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DOI：
10.1056/nejmoa040258
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发表时间：
2004-08-12
影响因子：
158.5
通讯作者：
Weissman, IL
中科院分区：
文献类型：
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作者：
Jamieson, CHM;Ailles, LE;Weissman, IL
背景：慢性髓性白血病（CML）的发展是由自我更新的白血病干细胞支持的。在正常小鼠造血干细胞中，自我更新的过程涉及(β)-连环蛋白信号通路。我们研究了CML中的白血病干细胞是否也使用(β)-catenin途径进行自我更新。方法：采用荧光活化细胞分选技术，分别从CML不同时期的骨髓和正常骨髓中分离造血干细胞、普通髓系祖细胞、粒细胞-巨噬细胞祖细胞和巨核细胞-红细胞祖细胞。BCR-ABL、(β)-连环蛋白和LEF-1转录物通过定量逆转录聚合酶链反应法在正常和CML造血干细胞和粒细胞巨噬细胞祖细胞中进行比较。用共聚焦荧光显微镜和淋巴细胞增强因子/ t细胞因子报告试验检测这些细胞中的核(β)-连环蛋白。体外复制实验用于鉴定自我更新细胞作为候选白血病干细胞，并通过(β)-catenin途径抑制剂轴蛋白（axin）慢病毒转导造血祖细胞来检测自我更新对(β)-catenin激活的依赖性。结果：原细胞危象和伊马替尼耐药CML患者的粒细胞-巨噬细胞祖细胞池扩大，表达BCR-ABL，与正常骨髓祖细胞相比，核(β)-连环蛋白水平升高。与正常的粒细胞-巨噬细胞祖细胞不同，CML粒细胞-巨噬细胞祖细胞形成自我更新、可重置的髓系集落，体外自我更新能力因轴蛋白的强制表达而降低。结论：CML粒细胞-巨噬细胞祖细胞中(β)-catenin的激活似乎增强了这些细胞的自我更新活性和白血病潜能。
Background: The progression of chronic myelogenous leukemia (CML) to blast crisis is supported by self-renewing leukemic stem cells. In normal mouse hematopoietic stem cells, the process of self-renewal involves the (beta)-catenin-signaling pathway. We investigated whether leukemic stem cells in CML also use the (beta)-catenin pathway for self-renewal.Methods: We used fluorescence-activated cell sorting to isolate hematopoietic stem cells, common myeloid progenitors, granulocyte-macrophage progenitors, and megakaryocyte-erythroid progenitors from marrow during several phases of CML and from normal marrow. BCR-ABL, (beta)-catenin, and LEF-1 transcripts were compared by means of a quantitative reverse-transcriptase-polymerase-chain-reaction assay in normal and CML hematopoietic stem cells and granulocyte-macrophage progenitors. Confocal fluorescence microscopy and a lymphoid enhancer factor/T-cell factor reporter assay were used to detect nuclear (beta)-catenin in these cells. In vitro replating assays were used to identify self-renewing cells as candidate leukemic stem cells, and the dependence of self-renewal on (beta)-catenin activation was tested by lentiviral transduction of hematopoietic progenitors with axin, an inhibitor of the (beta)-catenin pathway.Results: The granulocyte-macrophage progenitor pool from patients with CML in blast crisis and imatinib-resistant CML was expanded, expressed BCR-ABL, and had elevated levels of nuclear (beta)-catenin as compared with the levels in progenitors from normal marrow. Unlike normal granulocyte-macrophage progenitors, CML granulocyte-macrophage progenitors formed self-renewing, replatable myeloid colonies, and in vitro self-renewal capacity was reduced by enforced expression of axin.Conclusions: Activation of (beta)-catenin in CML granulocyte-macrophage progenitors appears to enhance the self-renewal activity and leukemic potential of these cells.