Array-based genomic resequencing of human leukemia
Array-based genomic resequencing of human leukemia
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DOI：
10.1038/onc.2010.117
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发表时间：
2010-06-24
期刊：
影响因子：
8
通讯作者：
Mano, H.
中科院分区：
文献类型：
--
作者：
Yamashita, Y.;Yuan, J.;Mano, H.
为了确定白血病中的癌基因，我们使用DNA序列阵列对白血病基因组进行了大规模的重新测序，这些DNA序列阵列确定了与5648个蛋白质编码基因的外显子或外显子-内含子边界相对应的类似9MBP的序列。来自急性髓系白血病(n=19)或骨髓增生性疾病(n=1)的CD34阳性原始细胞的基因组DNA杂交发现9148个非同义核苷酸改变。随后的分析表明，这些变化中的大多数也存在于配对的对照组的基因组DNA中，其中11个体细胞变化仅在白血病母细胞中发现。其中一个突变导致Janus kinase3(JAK3)511位氨基酸的Met-to-Ile替换，JAK3(M511I)蛋白在体外和体内都显示出转化潜力。进一步的JAK3突变筛查显示，在286例白血病患者中，共有9例出现了新的和已知的转化变化。我们的实验还显示，DNA甲基转移酶3A的882位氨基酸发生了由Arg到His的体细胞变化，导致DNA甲基化活性丧失50%。因此，我们的数据显示了人类白血病基因突变的独特特征。Oncogene(2010)29,3723-3731；doi：10.1038/onc.2010.117；2010年4月19日在线发布
To identify oncogenes in leukemias, we performed large-scale resequencing of the leukemia genome using DNA sequence arrays that determine similar to 9 Mbp of sequence corresponding to the exons or exon-intron boundaries of 5648 protein-coding genes. Hybridization of genomic DNA from CD34-positive blasts of acute myeloid leukemia (n = 19) or myeloproliferative disorder (n = 1) with the arrays identified 9148 nonsynonymous nucleotide changes. Subsequent analysis showed that most of these changes were also present in the genomic DNA of the paired controls, with 11 somatic changes identified only in the leukemic blasts. One of these latter changes results in a Met-to-Ile substitution at amino-acid position 511 of Janus kinase 3 (JAK3), and the JAK3(M511I) protein exhibited transforming potential both in vitro and in vivo. Further screening for JAK3 mutations showed novel and known transforming changes in a total of 9 out of 286 cases of leukemia. Our experiments also showed a somatic change responsible for an Arg-to-His substitution at amino-acid position 882 of DNA methyltransferase 3A, which resulted in a loss of DNA methylation activity of >50%. Our data have thus shown a unique profile of gene mutations in human leukemia. Oncogene (2010) 29, 3723-3731; doi: 10.1038/onc.2010.117; published online 19 April 2010