Serum microRNA profiles in children with autism

Mahesh Mundalil Vasu, Ayyappan Anitha, Ismail Thanseem, Katsuaki Suzuki, Kohei Yamada, Taro Takahashi, Tomoyasu Wakuda, Keiko Iwata, Masatsugu Tsujii, Toshirou Sugiyama, Norio Mori, Mahesh Mundalil Vasu, Ayyappan Anitha, Ismail Thanseem, Katsuaki Suzuki, Kohei Yamada, Taro Takahashi, Tomoyasu Wakuda, Keiko Iwata, Masatsugu Tsujii, Toshirou Sugiyama, Norio Mori

Abstract

Background: As regulators of gene expression, microRNAs (miRNAs) play a key role in the transcriptional networks of the developing human brain. Circulating miRNAs in the serum and plasma are remarkably stable and are suggested to have promise as noninvasive biomarkers for neurological and neurodevelopmental disorders. We examined the serum expression profiles of neurologically relevant miRNAs in autism spectrum disorder (ASD), a complex neurodevelopmental disorder characterized by multiple deficits in communication, social interaction and behavior.

Methods: Total RNA, including miRNA, was extracted from the serum samples of 55 individuals with ASD and 55 age- and sex-matched control subjects, and the mature miRNAs were selectively converted into cDNA. Initially, the expression of 125 mature miRNAs was compared between pooled control and ASD samples. The differential expression of 14 miRNAs was further validated by SYBR Green quantitative PCR of individual samples. Receiver-operating characteristic (ROC) analysis was used to evaluate the sensitivity and specificity of miRNAs. The target genes and pathways of miRNAs were predicted using DIANA mirPath software.

Results: Thirteen miRNAs were differentially expressed in ASD individuals compared to the controls. MiR-151a-3p, miR-181b-5p, miR-320a, miR-328, miR-433, miR-489, miR-572, and miR-663a were downregulated, while miR-101-3p, miR-106b-5p, miR-130a-3p, miR-195-5p, and miR-19b-3p were upregulated. Five miRNAs showed good predictive power for distinguishing individuals with ASD. The target genes of these miRNAs were enriched in several crucial neurological pathways.

Conclusions: This is the first study of serum miRNAs in ASD individuals. The results suggest that a set of serum miRNAs might serve as a possible noninvasive biomarker for ASD.

Keywords: Autism spectrum disorder; complementary DNA; microRNA; microarray; quantitative PCR.

Figures

Figure 1
Figure 1
Results from preliminary screening experiments. The Ct values obtained were normalized and calculated by three different strategies. miR-151a-3p, miR-181b-5p, miR-320a, miR-328, miR-433, miR-489, miR-572 and miR-663a were downregulated while miR-101-3p, miR-106b-5p, miR-19b-3p, miR-195-5p, miR-130a-3p and miR-27a-3p were upregulated. Bars represent the fold change in subjects with autism as compared to controls.
Figure 2
Figure 2
Scatter plot showing the differential expression of 13 microRNAs in autistic subjects.
Figure 3
Figure 3
Receiver operating characteristic (ROC) curve of 13 microRNAs. AUC, area under the ROC curve; SEN, sensitivity; SPE, specificity.
Figure 4
Figure 4
Neurologically relevant pathways (predicted by DIANA mirPath) of the target genes of differentially expressed microRNAs.

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Source: PubMed

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