High-throughput screening using iPSC-derived GABAergic neurons to elucidate disease-relevant phenotypes in serotonin 2A receptor gene variant-related sleep bruxism

使用 iPSC 衍生的 GABA 能神经元进行高通量筛选，以阐明与 5-羟色胺 2A 受体基因变异相关的睡眠磨牙症的疾病相关表型

• 批准号: 21K21049
• 负责人: サルカル アビジットクマール
• 金额: $ 1.91万
• 依托单位: Showa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Research Activity Start-up
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-08-30 至 2022-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21K21049/
• 关键词: iPSC-derived neurons; sleep bruxism; excitability; Sleep bruxism; iPS cell; GABAergic neurons; transcriptomic analysis; MEA array

The consequences of sleep bruxism (SB) appear to be serious orofacial pain and several dysfunction conditions, which seriously compromises the patient’s quality of life. However, the definitive mechanisms that promote SB are not well understood. We previously found a variant in the neuronal serotonin 2A receptor gene (HTR2A), rs6313 associated with the risk of SB and established human induced pluripotent stem cell (iPSC)-derived neurons from SB patients with this genetic variant. It has been suggested that attenuating activity of serotonin 2A receptor (5-HT2AR)-expressing GABAergic neurons during sleep may be involved in the mechanism of SB development. We found altered excitability in SB iPSC-derived neurons in the early stage of neurogenesis. This year we established two additional iPSC lines from a SB patient (SB3) and an unaffected control (C3) subject. In addition, we performed functional investigations of the SNP neurons at DIV31-51, 52-71, 72-91, and 92-111 of neurogenesis. We revealed that SB neurons showed significantly higher action potential firing frequency, higher gain, and shorter action potential half duration than control neurons over the course of DIV111 in culture. The altered electrophysiological properties of SB neurons indicate that affected cells may be hyperactive.

睡眠磨牙症（SB）的后果似乎是严重的口面部疼痛和几种功能障碍的条件，这严重损害了患者的生活质量。然而，促进SB的确切机制还不清楚。我们之前发现了神经元5-羟色胺2A受体基因（HTR 2A）的一个变体，rs6313与SB的风险相关，并从具有这种遗传变体的SB患者中建立了人类诱导多能干细胞（iPSC）衍生的神经元。研究表明，睡眠过程中表达5-HT 2A受体（5-HT 2AR）的GABA能神经元活性减弱可能参与SB的发生机制。我们发现SB iPSC衍生的神经元在神经发生的早期阶段的兴奋性改变。今年，我们从SB患者（SB 3）和未受影响的对照（C3）受试者中建立了两个额外的iPSC系。此外，我们在神经发生的DIV 31 -51、52-71、72-91和92-111进行了SNP神经元的功能研究。我们发现，SB神经元表现出显着更高的动作电位放电频率，更高的增益，和更短的动作电位半时程比对照神经元在培养过程中的DIV 111。SB神经元电生理特性的改变表明受影响的细胞可能是过度活跃的。

项目成果

Sleep bruxism iPSC-derived neurons display altered electrophysiology

睡眠磨牙症 iPSC 衍生的神经元表现出电生理学改变

• 发表时间: 2021
• 作者: Sarkar AK;Nakamura S;Nakai K;Abe Y;Hoashi Y;et al.
• 通讯作者: et al.

Electrophysiological characterization of sleep bruxism patient-specific iPSC-derived neurons

睡眠磨牙症患者特异性 iPSC 衍生神经元的电生理学特征

• 发表时间: 2021
• 作者: Sarkar AK;Nakamura S;Nakai K;Abe Y;Hoashi Y;et al.
• 通讯作者: et al.

Increased excitability of human iPSC-derived neurons in HTR2A variant-related sleep bruxism

HTR2A 变异相关的睡眠磨牙症中人类 iPSC 衍生神经元的兴奋性增加

• DOI: 10.1016/j.scr.2022.102658
• 发表时间: 2022
• 期刊: Stem Cell Research
• 影响因子: 1.2
• 作者: Sarkar Avijite Kumer;Nakamura Shiro;Nakai Kento;Sato Taro;Shiga Takahiro;Abe Yuka;Hoashi Yurie;Inoue Tomio;Akamatsu Wado;Baba Kazuyoshi
• 通讯作者: Baba Kazuyoshi

Postnatal Maturation of Glutamatergic Inputs onto Rat Jaw-closing and Jaw-opening Motoneurons

大鼠闭颌和张颌运动神经元谷氨酸能输入的出生后成熟

• DOI: 10.1016/j.neuroscience.2021.11.016
• 发表时间: 2022
• 期刊: Neuroscience
• 影响因子: 3.3
• 作者: Nakamura Shiro;Kajiwara Risa;Noguchi Tsuyoshi;Nakayama Kiyomi;Mochizuki Ayako;Dantsuji Masanori;Sarkar Avijite Kumer;Inoue Tomio
• 通讯作者: Inoue Tomio