Investigation of circulating serum microRNA-328-3p and microRNA-3135a expression as promising novel biomarkers for autism spectrum disorder

Abstract

Circulating microRNAs (miRNAs) are emerging as promising diagnostic biomarkers for autism spectrum disorder (ASD), but their usefulness for detecting ASD remains unclear. Nowadays, development of promising biomarkers for ASD remains a challenge. Recently, dysregulation of the miRNAs expression in postmortem brain tissue, serum and peripheral blood, have been associated with ASD. Circulating miRNAs are known to be secreted by a number of different cells and can interpose delivery of information into receiver cells, thus affecting their functions. Based on this fact, it is supposed that serum miRNAs could be a novel class of biomarkers for prognosis or diagnosis of pathological disorders including ASD. In the current research, we investigated whether the expression patterns of circulating miRNAs showed dysregulation in subjects diagnosed with ASD. Expression levels of serum miR-328-3p and miR-3135a were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) method of subjects diagnosed with ASD in comparison with healthy control subjects. Our data showed that miR-328-3p and miR-3135a were substantially down-regulated in ASD patients than in those of healthy control subjects. Moreover, target gene analysis of altered serum miRNAs displayed that these molecules targeted 162 genes denoted as unique validated targets in the miRWalk database, 71 of which appear to participate in biological pathways involved in synaptic pathways and neurodegenerative condition such as Alzheimer, Huntington and Parkinson diseases. Finally, the results strongly suggested that dys-regulated serum miRNAs might be involved in molecular pathways associated with ASD and miR-328-3p and miR-3135a have the potential to be promising novel biomarkers for ASD.

Circulating miRNAs, Small but Promising Biomarkers for Autism Spectrum Disorder

“…Although the research on ASD is progressively and exponentially growing, only 8 papers have explored the use of circulating miRNAs (ci-miRNAs) in biofluids (Table 1) such as in serum (Vasu et al, 2014; Cirnigliaro et al, 2017; Kichukova et al, 2017; Popov et al, 2018; Yu et al, 2018) and in saliva (Hicks et al, 2016, 2018, 2019; Figure 3). Moreover, from the few investigations, only the three studies performed using saliva specimens incorporated a high-throughput approach (RNA-seq).…”

“…The study of Popov et al (2018), showed differential expression changes of miR-3135a and miR-328-3p in the serum of ASD individuals using qRT-PCR technology. Remarkably these miRNAs were previously described to play a role in the etiopathogenesis of several neurological disorders (Provost, 2010; De Felice et al, 2012).…”

“…Their study showed that miR-3135a and miR-328-3p could discriminate between ASD individuals and controls as previously demonstrated by Kichukova et al (2017) being downregulated in their cohort. Using KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis, some of the target genes of miR-3135a and miR-328-3p are involved in neurodegenerative diseases, but their involvement in ASD is yet to be clarified (Popov et al, 2018).…”

Transcriptomic Analyses for Identification and Prioritization of Genes Associated With Alzheimer’s Disease in Humans

“…Moreover, this miRNA has been confirmed to dysregulate expression across the different brain tissue regions in AD patients (Puthiyedth et al, 2016). Hsa-mir-328 has been reported to be associated with a variety of neurologic disorders, such as autism spectrum disorder, Huntington's, Parkinson's, and Alzheimer's, and the biological functions of its targeting genes APP and BACE1 were validated experimentally in mouse brain tissues (Boissonneault et al, 2009;Provost, 2010;Nt et al, 2018). High-throughput experimental data from starBase were used to verify the results, and similar conclusions were obtained (see Supplementary Text S1 and Supplementary Table S7).…”

Enrichment and Correlation Analysis of Serum miRNAs in Comorbidity Between Arnold-Chiari and Tourette Syndrome Contribute to Clarify Their Molecular Bases

“…Numerous studies have shown that miRNAs affect cellular pathways and biological processes (Kim and Nam, 2006;Ragusa et al, 2017;O'Brien et al, 2018;Treiber et al, 2019), also playing an essential role in many aspects of neural development (e.g., neurogenesis, neuronal differentiation, maturation, synaptogenesis, neuronal plasticity, neuronal apoptosis) (Bian and Sun, 2011). Furthermore, miRNAs show specific expression and spatiotemporal patterns in mammalian central nervous systems (CNS) (Kosik, 2006;Rao et al, 2013); alterations of miRNA expression have been associated with various brain anomalies and neuropsychiatric disorders (Xu et al, 2010(Xu et al, , 2012Mundalil Vasu et al, 2014;Wu et al, 2015;Hicks et al, , 2019Alural et al, 2017;Cirnigliaro et al, 2017;Kichukova et al, 2017;Nt et al, 2018;Zadehbagheri et al, 2019), delineating a discriminative molecular signature (Salta and De Strooper, 2012). Furthermore, the evidence that extracellular RNA can be detected in all mammalian body fluids, including peripheral blood (Kosaka et al, 2010;Turchinovich et al, 2011Turchinovich et al, , 2013, opened the way to an important source of biomarkers (Rao et al, 2013;Kichukova et al, 2015;Rizzo et al, 2015): this finding revealed the possibility to analyze the expression of circulating miRNAs (cmiRNAs) through liquid biopsies, in a minimally invasive way (Rao et al, 2013;Nevel et al, 2018).…”