Insertional mutagenesis identifies genes that promote the immortalization of primary bone marrow progenitor cells
Insertional mutagenesis identifies genes that promote the immortalization of primary bone marrow progenitor cells
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DOI：
10.1182/blood-2005-03-1113
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发表时间：
2005-12-01
期刊：
影响因子：
20.3
通讯作者：
Copeland, NG
中科院分区：
文献类型：
--
作者：
Du, Y;Jenkins, NA;Copeland, NG
逆转录病毒通过插入诱变肿瘤基因诱导造血疾病，为肿瘤基因的发现提供了有价值的分子标签。在这里，我们表明插入突变也可以识别促进造血细胞永生的基因，造血细胞通常只有有限的自我更新。用仅表达neo的复制无能小鼠干细胞病毒（MSCV）转导小鼠骨髓细胞，然后在含有干细胞因子（SCIF）和白细胞介素-3 （IL-3）的液体培养基中连续传代，在约50%的感染培养物中产生具有中性粒细胞和巨噬细胞分化潜力的永生化未成熟骨髓细胞系。超过一半的品系在Evil或Prdm16上有MSCV插入。这些基因座编码转录因子同源物，是经过验证的人髓性白血病基因。整合位于内含子1或2中，它们促进缺乏PRDI-BF1-RIZ1同源（PR）结构域的截断蛋白的表达，类似于在具有EVI1或PRDM16突变的人类白血病中观察到的情况。Evi1过表达似乎足以使未成熟骨髓细胞永生，似乎不需要任何其他协同突变。通过插入突变鉴定的基因也可能参与白血病干细胞的永生，因此代表了治疗癌症的有吸引力的药物靶点。
Retroviruses can induce hematopoietic disease via insertional mutagenesis of cancer genes and provide valuable molecular tags for cancer gene discovery. Here we show that insertional mutagenesis can also identify genes that promote the immortalization of hematopoietic cells, which normally have only limited self-renewal. Transduction of mouse bone marrow cells with replication-incompetent murine stem cell virus (MSCV) expressing only neo, followed by serial passage in liquid culture containing stem cell factor (SCIF) and interleukin-3 (IL-3), produced immortalized immature myeloid cell lines with neutrophil and macrophage differentiation potential in about 50% of the infected cultures. More than half of the lines have MSCV insertions at Evil or Prdm16. These loci encode transcription factor homologs and are validated human myeloid leukemia genes. Integrations are located in intron 1 or 2, where they promote expression of truncated proteins lacking the PRDI-BF1-RIZ1 homologous (PR) domain, similar to what is observed in human leukemias with EVI1 or PRDM16 mutations. Evi1 overexpression alone appears sufficient to immortalize immature myeloid cells and does not seem to require any other cooperating mutations. Genes identified by insertional mutagenesis by their nature could also be involved in immortalization of leukemic stem cells, and thus represent attractive drug targets for treating cancer.