AlShawaf Abdullah J, AlNassar Sarah A, AlGhamdi Norah, Mattei Cristiana, Lim Shiang Y, Dottori Mirella, Al-Mohanna Futwan A
Department of Physical Therapy, Mohammed Al-Mana College for Medical Sciences, Dammam 34222, Saudi Arabia.
Department of Physiological Sciences, College of Medicine, AlFaisal University, Riyadh 11533, Saudi Arabia.
Cells. 2025 Sep 8;14(17):1402. doi: 10.3390/cells14171402.
Autism Spectrum Disorder (ASD) is a neurodevelopmental condition that affects communication, social interaction, and behavior. Calcium (Ca) signaling dysregulation has been frequently highlighted in genetic studies as a contributing factor to aberrant developmental processes in ASD. Herein, we used ASD and control induced pluripotent stem cells (iPSCs) to investigate transcriptomic and functional Ca dynamics at various stages of differentiation to cortical neurons. Idiopathic ASD and control iPSC lines underwent the dual SMAD inhibition differentiation protocol to direct their fate toward cortical neurons. Samples from multiple time points along the course of differentiation were processed for bulk RNA sequencing, spanning the following sequential stages: the iPSC stage, neural induction (NI) stage, neurosphere (NSP) stage, and differentiated cortical neuron (Diff) stage. Our transcriptomic analyses suggested that the numbers of Ca signaling-relevant differentially expressed genes between ASD and control samples were higher in the iPSC and Diff stages. Accordingly, samples from the iPSC and Diff stages were processed for Ca imaging studies. Results revealed that iPSC-stage ASD samples displayed elevated maximum Ca levels in response to ATP compared to controls. By contrast, in the Diff stage, ASD neurons showed reduced maximum Ca levels in response to ATP but increased maximum Ca levels in response to KCl and DHPG relative to controls. Considering the distinct functional signaling contexts of these stimuli, this differential profile of receptor- and ionophore-mediated Ca response suggests that aberrant calcium homeostasis underlies the pathophysiology of ASD neurons. Our data provides functional evidence for Ca signaling dysregulation during neurogenesis in idiopathic ASD.
自闭症谱系障碍（ASD）是一种影响沟通、社交互动和行为的神经发育疾病。钙（Ca）信号失调在遗传研究中经常被强调为ASD异常发育过程的一个促成因素。在此，我们使用ASD和对照诱导多能干细胞（iPSC）来研究向皮质神经元分化的各个阶段的转录组和功能性钙动力学。特发性ASD和对照iPSC系采用双SMAD抑制分化方案，将其命运导向皮质神经元。沿着分化过程的多个时间点的样本进行了批量RNA测序，涵盖以下连续阶段：iPSC阶段、神经诱导（NI）阶段、神经球（NSP）阶段和分化的皮质神经元（Diff）阶段。我们的转录组分析表明，ASD和对照样本之间与钙信号相关的差异表达基因数量在iPSC和Diff阶段更高。因此，对iPSC和Diff阶段的样本进行了钙成像研究。结果显示，与对照相比，iPSC阶段的ASD样本对ATP的最大钙水平升高。相比之下，在Diff阶段，ASD神经元对ATP的最大钙水平降低，但对KCl和DHPG的最大钙水平相对于对照增加。考虑到这些刺激的不同功能信号背景，这种受体和离子载体介导的钙反应的差异表明，异常的钙稳态是ASD神经元病理生理学的基础。我们的数据为特发性ASD神经发生过程中的钙信号失调提供了功能证据。
