Hosokawa Rie, Yoshino Yuta, Funahashi Yu, Horiuchi Fumie, Iga Jun-Ichi, Ueno Shu-Ichi
Department of Neuropsychiatry, Molecules and Function, Ehime University Graduate School of Medicine, Shitsukawa, Toon 791-0295, Ehime, Japan.
Center for Child Health, Behavior and Development, Ehime University Hospital, Toon 791-0295, Ehime, Japan.
Brain Sci. 2022 Dec 22;13(1):27. doi: 10.3390/brainsci13010027.
Autism spectrum disorder (ASD), is a neurodevelopmental disorder that is known to have a high degree of heritability. Diagnosis of ASD is difficult because of the high heterogeneity of the clinical symptoms. MicroRNAs (miRNAs) can potentially be diagnostic biomarkers for ASD, and several studies have shown the relationship between miRNAs and ASD pathogenesis. In this study, we investigated ten miRNA and mRNA expression of target genes in peripheral blood to explore a diagnostic biomarker for ASD.
We recruited control and ASD subjects for the discovery cohort ( = 6, each) and replication cohort ( = 20, each). Using qPCR, miRNA and mRNA expression was measured using the SYBR green and probe methods, respectively. In-silico prediction was used for identifying target genes of miRNAs. An in vitro experiment using HEK293 cells was conducted to investigate whether miR-15b-5p modulates the predicted target genes (TGFBR3 and MYBL1).
miR-15b-5p expression indicated an increased trend in the discovery cohort ( = 0.052) and a significant upregulation in the replication cohort ( = 0.021). In-silico analysis revealed that miR-15b-5p is relevant to cell development and Wnt signaling. The decreased trends of TGFBR3 and MYBL expression were the same as in previous RNA-seq data. MiR-15b-5p positively regulated TGFBR3 expression in in vitro experiments.
Upregulated miR-15b-5p expression may represent a useful diagnostic marker of ASD subjects, and it may regulate TGFBR3 mRNA expression. These findings indicate a new perspective in the understanding of the pathogenesis of ASD.
自闭症谱系障碍（ASD）是一种神经发育障碍，具有高度遗传性。由于临床症状高度异质性，ASD的诊断较为困难。微小RNA（miRNA）有可能成为ASD的诊断生物标志物，多项研究已表明miRNA与ASD发病机制之间的关系。在本研究中，我们调查了外周血中10种miRNA及其靶基因的mRNA表达，以探索ASD的诊断生物标志物。
我们招募了对照组和ASD受试者作为发现队列（每组n = 6）和验证队列（每组n = 20）。使用qPCR，分别采用SYBR Green法和探针法测量miRNA和mRNA表达。利用计算机模拟预测来鉴定miRNA的靶基因。进行了一项使用HEK293细胞的体外实验，以研究miR-15b-5p是否调节预测的靶基因（TGFBR3和MYBL1）。
miR-15b-5p表达在发现队列中呈上升趋势（P = 0.052），在验证队列中显著上调（P = 0.021）。计算机模拟分析表明，miR-15b-5p与细胞发育和Wnt信号通路相关。TGFBR3和MYBL表达的下降趋势与先前的RNA-seq数据一致。在体外实验中，miR-15b-5p正向调节TGFBR3表达。
miR-15b-5p表达上调可能是ASD受试者的一种有用诊断标志物，并且它可能调节TGFBR3 mRNA表达。这些发现为理解ASD的发病机制提供了新的视角。
