Generation of neural cells using iPSCs from sleep bruxism patients with 5-HT2A polymorphism
Generation of neural cells using iPSCs from sleep bruxism patients with 5-HT2A polymorphism
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DOI：
10.1016/j.jpor.2016.11.003
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发表时间：
2017-07-01
影响因子：
3.6
通讯作者：
Baba, Kazuyoshi
中科院分区：
文献类型：
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作者：
Hoashi, Yurie;Okamoto, Satoshi;Baba, Kazuyoshi
目的：睡眠磨牙症（Sleep bruxism，SB）是一种与睡眠相关的运动障碍，其特征是睡眠期间牙齿的磨牙和紧咬，导致牙齿异常磨损和牙齿或牙根断裂等多种临床问题。我们以前的研究确定了5-羟色胺2A受体（5-HT 2A）单核苷酸多态性（SNP）rs6313 C>T与SB的基因组关联，其中C等位基因携带者与SB风险增加4.25倍相关。基于这一发现，本研究的目的是使用SB患者特异性诱导多能干细胞（iPSC）产生神经细胞。采用多导睡眠图和TaqMan基因分型技术对2例SB患者和2例对照者进行rs6313基因C/C型和T/T型的筛查。通过引入重编程因子，从外周血单核细胞建立了四个iPSC系，两个来自SB患者，两个来自对照。我们使用未分化多能细胞标记物的表达、多能性标记物的免疫染色、三胚层测定和核型分析对iPSC进行质量控制测定。使用神经球培养系统将建立的iPSCs分化为神经元。结果：患者特异性诱导的多能干细胞成功分化为表达5-HT 2A的神经元。结论：这份报告是第一次使用来自5-HT 2A多态性睡眠磨牙症患者的iPSC成功产生神经细胞，这有可能阐明SB的病因和潜在机制。（C）2016日本口腔修复学会。由爱思唯尔有限公司出版。保留所有权利。
Purpose: Sleep bruxism (SB) is classified as a sleep-related movement disorder characterized by grinding and clenching of the teeth during sleep, which is responsible for a variety of clinical problems such as abnormal tooth attrition and fracture of teeth or roots. Little is known about the etiology of SB. Our previous study identified a genomic association of the serotonin 2A receptor (5-HT2A) single nucleotide polymorphism (SNP), rs6313 C>T, with SB, where the C allele carrier is associated with a 4.25-fold increased risk of SB. Based on this finding, the aim of this study was to generate of neural cells using SB patient-specific induced pluripotent stem cells (iPSCs).Methods: Two SB patients with C/C genotype of rs6313 and two controls with T/T genotype were screened by laboratory-based polysomnographic recordings and the TaqMan genotyping assay. Four lines of iPSCs, two from SB patients and two from controls, were established from peripheral blood mononuclear cells by introduction of reprogramming factors. We performed quality control assays on iPSCs using expression of markers for undifferentiated pluripotent cells, immunostaining for pluripotency markers, a three-germ layer assay, and karyotype analysis. The established iPSCs were differentiated into neurons using the neurosphere culture system. 5-HT2A gene expression in these neurons was evaluated by quantitative real-time PCR.Results: Patient-specific iPSCs were successfully differentiated into neurons expressing 5-HT2A.Conclusions: This report is the first successful generation of neural cells using iPSCs from sleep bruxism patients with 5-HT2A polymorphism, which has the potential to elucidate the etiology and underlying mechanism of SB. (C) 2016 Japan Prosthodontic Society. Published by Elsevier Ltd. All rights reserved.