Mooney Catherine, Parlante Andrea, Canarutto Giulia, Grigoli Andrea, Scattoni Maria Luisa, Ricceri Laura, Jimenez-Mateos Eva Maria, Sanz-Rodriguez Amaya, Clementi Elena, Piazza Silvano, Henshall David C, Provenzano Giovanni
Department of Physiology & Medical Physics, RCSI University of Medicine and Health Sciences, Dublin, Ireland.
School of Computer Science, University College Dublin, Dublin, Ireland.
Mol Neurobiol. 2025 Apr 14. doi: 10.1007/s12035-025-04900-x.
Autism spectrum disorder (ASD) is a neurodevelopmental condition caused by both genetic and environmental factors. Since no single gene variant accounts for more than 1% of the cases, the converging actions of ASD-related genes and other factors, including microRNAs (miRNAs), may contribute to ASD pathogenesis. To date, few studies have simultaneously investigated the mRNA and miRNA profiles in an ASD-relevant model. The BTBR mouse strain displays a range of behaviors with ASD-like features but little is known about the protein-coding and noncoding gene expression landscape that may underlie the ASD-like phenotype. Here we performed parallel mRNA and miRNA profiling using the prefrontal cortex (PFC) of BTBR and C57BL/6 J (B6) mice. This identified 1063 differentially expressed genes and 48 differentially expressed miRNAs. Integration of mRNA and miRNA data identified a strong inverse relationship between upregulated (DEGs) and downregulated miRNAs, and vice versa. Pathway analysis, taking account of the inverse relationship between differentially expressed miRNAs and their target mRNAs highlighted significant shared enrichment in immune signaling, myelination, and neurodevelopmental processes. Notably, miRNA changes were predicted to affect synapse-related functions but we did not find enrichment of protein-coding genes linked to cellular components or biological processes related to synapses in the PFC of BTBR mice, indicating processes may evade miRNA control. In contrast, other miRNAs were predicted to have extensive relationships with DEGs suggesting their role as potential hub coordinators of gene expression. Profiling findings were confirmed via qRT-PCR for representative protein-coding transcripts and miRNAs. Our study underscores the complex interplay between gene expression and miRNA regulation within immune and inflammatory pathways in the BTBR model, offering insights into the neurodevelopmental mechanisms of ASD. These results support the value of the BTBR mouse model and identify strategies that could adjust molecular pathways for therapeutic applications in ASD research.
自闭症谱系障碍(ASD)是一种由遗传和环境因素共同导致的神经发育疾病。由于没有单一基因变异能解释超过1%的病例,与ASD相关的基因及其他因素(包括微小RNA,即miRNA)的共同作用可能导致了ASD的发病机制。迄今为止,很少有研究在与ASD相关的模型中同时研究mRNA和miRNA谱。BTBR小鼠品系表现出一系列类似ASD的行为特征,但对于可能构成类似ASD表型基础的蛋白质编码和非编码基因表达情况知之甚少。在此,我们使用BTBR和C57BL/6 J(B6)小鼠的前额叶皮质(PFC)进行了平行的mRNA和miRNA分析。这确定了1063个差异表达基因和48个差异表达miRNA。mRNA和miRNA数据的整合确定了上调的(差异表达基因,即DEGs)和下调的miRNA之间存在强烈的负相关关系,反之亦然。通路分析考虑到差异表达的miRNA与其靶标mRNA之间的负相关关系,突出了免疫信号传导、髓鞘形成和神经发育过程中显著的共同富集。值得注意的是,预测miRNA的变化会影响与突触相关的功能,但我们在BTBR小鼠的PFC中未发现与细胞成分或与突触相关的生物学过程相关的蛋白质编码基因富集,这表明这些过程可能逃避了miRNA的调控。相比之下,其他miRNA被预测与差异表达基因有广泛的关系,表明它们作为基因表达潜在枢纽协调者的作用。通过qRT-PCR对代表性的蛋白质编码转录本和miRNA进行分析,证实了分析结果。我们的研究强调了BTBR模型中免疫和炎症通路内基因表达与miRNA调控之间的复杂相互作用,为ASD的神经发育机制提供了见解。这些结果支持了BTBR小鼠模型的价值,并确定了可调整分子通路以用于ASD研究治疗应用的策略。