Development of a Fluorescence in Situ Hybridization Probe for Detecting IKZF1 Deletion Mutations in Patients with Acute Lymphoblastic Leukemia
Development of a Fluorescence in Situ Hybridization Probe for Detecting IKZF1 Deletion Mutations in Patients with Acute Lymphoblastic Leukemia
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开发用于检测急性淋巴细胞白血病患者 IKZF1 缺失突变的荧光原位杂交探针
DOI：
10.1016/j.jmoldx.2018.02.005
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Teshima Takanori
中科院分区：
文献类型：
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作者：
Hashiguchi Junichi;Onozawa Masahiro;Oguri Satoshi;Fujisawa Shinichi;Tsuji Masahisa;Okada Kohei;Nakagawa Masao;Hashimoto Daigo;Kahata Kaoru;Kondo Takeshi;Shimizu Chikara;Teshima Takanori
fikzf1基因内缺失是急性淋巴细胞白血病中一种复发性基因组改变。这些缺失是由非法变量（多样性）通过隐式重组信号序列（rss）加入重组介导的。我们开发了一种荧光原位杂交（FISH）探针集，可以检测任何类型的fikzf1缺失，包括常见缺失的外显子4至7区域。该探针组由设计的用于常见缺失区域的探针（Cy3，红色）和用于检测3 '侧翼区域的细菌人工染色体克隆探针（光谱绿色）组成。intactikzf1显示融合信号，被删除的等位基因显示红色信号（0R1G1F）丢失。FISH探针对人类白血病细胞系和临床样品工作正确。1例显示非典型分离信号（1R1G1F）。该病例的反相PCR显示，excisedikzf1片段重排到2号染色体上Ig κ的合法RSS位点，表明该重组激活基因1/2介导的事件具有致病作用。在这项研究中，我们建立了一种快速、定量和经济有效的方式检测ikzf1缺失的FISH探针，结果为通过重排一个隐式rss介导的基因组片段在急性淋巴细胞白血病病理中进行b细胞受体编辑提供了新的见解。
Intragenic deletion ofIKZF1is a recurrent genomic alteration in acute lymphoblastic leukemia. The deletions are mediated by illegitimate variable(diversity)joining recombination via cryptic recombination signal sequences (RSSs). We developed a fluorescencein situhybridization (FISH) probe set that can detect any type ofIKZF1deletion, including the commonly deleted exon 4 to 7 region. The probe set consists of a designed probe for the commonly deleted region (Cy3; red) and a bacterial artificial chromosomes clone probe for detecting the 3′ flanking region (Spectrum Green). IntactIKZF1showed a fusion signal, and the deleted allele showed loss of the red signal (0R1G1F). The FISH probes worked correctly for human leukemic cell lines and clinical samples. One case showed an atypical break-apart signal (1R1G1F). Inverse PCR of the case revealed rearrangement of the excisedIKZF1fragment into a legitimate RSS site at Ig κ on chromosome 2, suggesting a pathogenic role of this recombination-activating gene 1/2-mediated event. In this study, we established FISH probe detectingIKZF1deletion in a quick, quantitative, and cost-effective manner, and the results provided a novel insight into B-cell receptor editing by rearrangement of a cryptic RSS-mediated genomic fragment in acute lymphoblastic leukemia pathology.