Circulating miRNome profiling in Moyamoya disease-discordant monozygotic twins and endothelial microRNA expression analysis using iPS cell line

Abstract

BackgroundMoyamoya disease (MMD) is characterized by progressive stenosis of intracranial arteries in the circle of Willis with unknown etiology even after the identification of a Moyamoya susceptible gene, RNF213. Recently, differences in epigenetic regulations have been investigated by a case-control study in MMD. Here, we employed a disease discordant monozygotic twin-based study design to unmask potential confounders.MethodsCirculating genome-wide microRNA (miRNome) profiling was performed in MMD-discordant monozygotic twins, non-twin-MMD patients, and non-MMD healthy volunteers by microarray followed by qPCRvalidation, using blood samples. Differential plasma-microRNAs were further quantified in endothelial cells differentiated from iPS cell lines (iPSECs) derived from another independent non-twin cohort. Lastly, their target gene expression in the iPSECs was analyzed.ResultsMicroarray detected 309 plasma-microRNAs in MMD-discordant monozygotic twins that were also detected in the non-twin cohort. Principal component analysis of the plasma-microRNA expression level demonstrated distinct 2 groups separated by MMD and healthy control in the twin- and non-twin cohorts. Of these, differential upregulations of hsa-miR-6722-3p/− 328-3p were validated in the plasma of MMD (absolute log2 expression fold change (logFC) > 0.26 for the twin cohort; absolute logFC > 0.26, p < 0.05, and q < 0.15 for the non-twin cohort). In MMD derived iPSECs, hsa-miR-6722-3p/− 328-3p showed a trend of up-regulation with a 3.0- or higher expression fold change. Bioinformatics analysis revealed that 41 target genes of miR-6722-3p/− 328-3p were significantly down-regulated in MMD derived iPSECs and were involved in STAT3, IGF-1-, and PTEN-signaling, suggesting a potential microRNA-gene expression interaction between circulating plasma and endothelial cells.ConclusionsOur MMD-discordant monozygotic twin-based study confirmed a novel circulating microRNA signature in MMD as a potential diagnostic biomarker minimally confounded by genetic heterogeneity. The novel circulating microRNA signature can contribute for the future functional microRNA analysis to find new diagnostic and therapeutic target of MMD.Electronic supplementary materialThe online version of this article (10.1186/s12920-018-0385-3) contains supplementary material, which is available to authorized users.

Integrated analysis of DNA methylation and transcriptome profiling of polycystic ovary syndrome

“…Statistical P-values were obtained by Limma package and adjusted into the false discovery rate (FDR) using the Benjamini-Hochberg method (16). The thresholds for screening DEGs and DMGs were set as FDR<0.05 and |log 2 fold-change (FC)|>0.263 (17).…”

The Impact of Moyamoya Disease and RNF213 Mutations on the Spectrum of Plasma Protein and MicroRNA

“…Several MMD-associated miRNAs in whole plasma, EVs and EV-depleted plasma were identified. Interestingly, there are few overlapping miRNAs with prior studies on moyamoya-associated circulating miRNA changes [39,40]. This difference was probably caused by different sample types, measurement platforms and NGS library construction methods used in this study.…”

“…This difference was probably caused by different sample types, measurement platforms and NGS library construction methods used in this study. In a study characterizing MMD plasma samples by microarray and qPCR, the authors validated two up-regulated miRNAs, hsa-miR-6722-3p and hsa-miR-328-3p [40]. Our sequencing analysis did not detect miR-6722-3p, but miR-328-3p showed an increased concentration in EV-depleted plasma from MMD patients (Table 2).…”

Pathological Circulating Factors in Moyamoya Disease

“…Similarly, hsa-miR-6722-3p/-328-3p, interacting with endothelial cell by having an impact on STAT3 (signal transducer and activator of transcription 3,) IGF-1 (insulin Like Growth Factor 1), and PTEN (phosphatase and tensin homolog)-signaling pathways, were abnormally upregulated in an MMD-discordant, monozygotic twin cohort. [ 50 ] The EV (vesicle encapsulated) miRNA may also take part in the pathology of MMD by affecting the signaling pathway of angiogenesis [ 51 ]. Regulating endothelial function and angiogenesis, the specific circulating miRNA aberrant in MMD patients is not only a potential therapeutic target and diagnostic biomarker, but also a key to better understanding a thorough pathological mechanism in MMD.…”