Analysis of induced pluripotent stem cell clones derived from a patient with mosaic neurofibromatosis type 2
Analysis of induced pluripotent stem cell clones derived from a patient with mosaic neurofibromatosis type 2
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2 型镶嵌神经纤维瘤病患者诱导多能干细胞克隆的分析
DOI：
10.1002/ajmg.a.62700
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发表时间：
2022
影响因子：
2
通讯作者：
Fujimura Miki
中科院分区：
文献类型：
--
作者：
Ishi Yukitomo;Era Takumi;Yuzawa Sayaka;Okamoto Michinari;Sawaya Ryosuke;Motegi Hiroaki;Yamaguchi Shigeru;Terasaka Shunsuke;Houkin Kiyohiro;Fujimura Miki
由于2型神经纤维瘤病（NF 2）亚型变异等位基因频率低，嵌合体的诊断具有挑战性。在这项研究中，我们产生了诱导多能干细胞（iPSC），从临床诊断为NF2的患者的基础上多个神经鞘瘤，包括双侧前庭神经鞘瘤和脑膜瘤。对患者外周血单个核细胞（MNCs）的遗传分析未能检测到NF 2改变，但在所有单独的肿瘤中成功发现了p.Q65X（c.193C>T）突变，其中3例颅内脑膜瘤和1例眶内神经鞘瘤，并使用深度测序证实了NF 2改变的嵌合体诊断。从外周血中的MNC建立了具有患者来源的iPSC的五个不同克隆，其显示多能标志物的充分表达。遗传分析显示，从MNC产生的五个iPSC系中的一个具有与NF 2中发现的相同的p.Q65X突变。在具有野生型和突变型NF2的iPSC克隆之间，与NF2相关的基因的表达没有显著差异。在这种情况下，单核骨髓源性干细胞的克隆扩增概括了NF 2中嵌合体的遗传改变。来自嵌合型NF2的患者来源的iPSC将有助于进一步研究NF2改变的功能。
The diagnosis of mosaicism is challenging in patients with neurofibromatosis type 2 (NF2) subset due to low variant allele frequency. In this study, we generated induced pluripotent stem cells (iPSCs) were generated from a patient clinically diagnosed with NF2 based on multiple schwannomas, including bilateral vestibular schwannomas and meningiomas. Genetic analysis of the patient's mononuclear cells (MNCs) from peripheral blood failed to detectNF2alteration but successfully found p.Q65X (c.193C>T) mutation in all separate tumors with three intracranial meningiomas and one intraorbital schwannoma, and confirming mosaicism diagnosis inNF2alteration using deep sequencing. Five different clones with patient‐derived iPSCs were established from MNCs in peripheral blood, which showed sufficient expression of pluripotent markers. Genetic analysis showed that one of five generated iPSC lines from MNCs had the same p.Q65X mutation as that found inNF2. There was no significant difference in the expression of genes related toNF2between iPSC clones with the wild‐type and mutantNF2. In this case, clonal expansion of mononuclear bone marrow‐derived stem cells recapitulated mosaicism's genetic alteration in NF2. Patient‐derived iPSCs from mosaic NF2 would contribute to further functional research ofNF2alteration.