Zhao Sha, Zhong Yan, Shen Fang, Cheng Xinning, Qing Xiaojuan, Liu Jiamin
Department of Child Healthcare, Hunan Children's Hospital, Changsha, China; University of South China, Hengyang, China.
Pediatrics Research Institute of Hunan Province, Hunan Children's Hospital, Changsha, China.
Biomol Biomed. 2024 Mar 11;24(2):292-301. doi: 10.17305/bb.2023.9552.
Exosomes have been demonstrated to exert momentous roles in autism spectrum disorder (ASD). However, few studies have reported a correlation between exosomal microRNAs (miRNAs) and ASD. To date, our understanding of crucial competing endogenous RNA (ceRNA) networks in ASD remains limited. Herein, the exosomal miRNA profile in the peripheral blood of children with ASD and healthy controls was investigated and the level of immune cell infiltration in ASD was evaluated to determine the distribution of immune cell subtypes. Exosomes were isolated from the peripheral blood of ten children with ASD and ten healthy controls, and further identified using transmission electron microscopy and western blot analysis. RNA sequencing was conducted to investigate exosomal miRNA profiles in patients with ASD. The mRNA and circular RNA (circRNA) expression profiles were acquired from the Gene Expression Omnibus (GEO) database. Differentially expressed mRNAs (DEmRNAs), miRNAs (DEmiRNAs), and circRNAs (DEcircRNAs) were identified and ceRNA regulatory networks were constructed. Furthermore, the immune cell infiltration levels in patients with ASD were evaluated. Exosomes were spherical, approximately 100 nm in size, and were confirmed via western blot analysis using exosome-associated markers CD9, CD63, and CD81. Thirty-five DEmRNAs, 63 DEmiRNAs, and 494 DEcircRNAs were identified in patients with ASD. CeRNA regulatory networks, including 6 DEmRNAs, 14 DEmiRNAs, and 86 DEcircRNAs, were established. Correlation analysis indicated that leucine-rich glioma inactivated protein 1 (LGI1) expression was significantly positively correlated with the content of CD8+ T cells. Our findings may be conducive to offering novel insights into this disease and providing further evidence of transcriptomic abnormalities in ASD.
外泌体已被证明在自闭症谱系障碍(ASD)中发挥着重要作用。然而,很少有研究报道外泌体微小RNA(miRNA)与ASD之间的相关性。迄今为止,我们对ASD中关键的竞争性内源性RNA(ceRNA)网络的了解仍然有限。在此,研究了ASD儿童和健康对照外周血中的外泌体miRNA谱,并评估了ASD中的免疫细胞浸润水平,以确定免疫细胞亚型的分布。从10名ASD儿童和10名健康对照的外周血中分离出外泌体,并使用透射电子显微镜和蛋白质印迹分析进一步鉴定。进行RNA测序以研究ASD患者的外泌体miRNA谱。从基因表达综合数据库(GEO)中获取mRNA和环状RNA(circRNA)表达谱。鉴定了差异表达的mRNA(DEmRNA)、miRNA(DEmiRNA)和circRNA(DEcircRNA),并构建了ceRNA调控网络。此外,评估了ASD患者的免疫细胞浸润水平。外泌体呈球形,大小约为100nm,并通过使用外泌体相关标志物CD9、CD63和CD81的蛋白质印迹分析得到证实。在ASD患者中鉴定出35个DEmRNA、63个DEmiRNA和494个DEcircRNA。建立了包括6个DEmRNA、14个DEmiRNA和86个DEcircRNA的ceRNA调控网络。相关性分析表明,富含亮氨酸的胶质瘤失活蛋白1(LGI1)表达与CD8 + T细胞含量显著正相关。我们的研究结果可能有助于为这种疾病提供新的见解,并为ASD转录组异常提供进一步的证据。