Background Autism spectrum disorder (ASD) is a neurodevelopmental disorder with a high co-morbidity of epilepsy and associated with hundreds of rare risk factors. NRXN1 deletion is among the commonest rare genetic factors shared by ASD, schizophrenia, intellectual disability, epilepsy, and developmental delay. However, how NRXN1 deletions lead to different clinical symptoms is unknown. Patient-derived cells are essential to investigate the functional consequences of NRXN1 lesions to human neurons in different diseases. Methods Skin biopsies were donated by five healthy donors and three ASD patients carrying NRXN1α +/- deletions. Seven control and six NRXN1α +/- iPSC lines were derived and differentiated into day 100 cortical excitatory neurons using dual SMAD inhibition. Calcium (Ca2+) imaging was performed using Fluo4-AM, and the properties of Ca2+ transients were compared between two groups of neurons. Transcriptome analysis was carried out to undercover molecular pathways associated with NRXN1α +/- neurons. Results NRXN1α +/- neurons were found to display altered calcium dynamics, with significantly increased frequency, duration, and amplitude of Ca2+ transients. Whole genome RNA sequencing also revealed altered ion transport and transporter activity, with upregulated voltage-gated calcium channels as one of the most significant pathways in NRXN1α +/- neurons identified by STRING and GSEA analyses. Conclusions This is the first report to show that human NRXN1α +/- neurons derived from ASD patients' iPSCs present novel phenotypes of upregulated VGCCs and increased Ca2+ transients, which may facilitate the development of drug screening assays for the treatment of ASD.

中文翻译：

---

从ASD诱导的多能干细胞衍生的NRXN1α+/-神经元中增加的Ca2 +信号传导。

背景自闭症谱系障碍（ASD）是一种神经发育障碍，具有高度的癫痫合并症，并与数百种罕见的危险因素相关。NRXN1缺失是ASD，精神分裂症，智力残疾，癫痫和发育迟缓所共有的最常见的罕见遗传因素之一。但是，未知NRXN1删除如何导致不同的临床症状。患者来源的细胞对于研究NRXN1病变对不同疾病中的人类神经元的功能后果至关重要。方法由五名健康捐献者和三名携带NRXN1α+/-缺失的ASD患者捐献皮肤活检。使用双重SMAD抑制，获得了七个对照和六个NRXN1α+/- iPSC系，并将其分化为100天皮质兴奋性神经元。使用Fluo4-AM进行钙（Ca2 +）成像，比较了两组神经元之间的Ca2 +瞬变特性。进行转录组分析以发现与NRXN1α+/-神经元相关的分子途径。结果发现NRXN1α+/-神经元显示钙动力学改变，Ca2 +瞬变的频率，持续时间和幅度显着增加。全基因组RNA测序还揭示了离子转运和转运蛋白活性的改变，电压门控钙通道的上调是STRING和GSEA分析所鉴定的NRXN1α+/-神经元中最重要的途径之一。结论这是第一份表明ASD患者的iPSC衍生的人NRXN1α+/-神经元表现出上调的VGCC的新表型和增加的Ca2 +瞬变的报告，