MiR-455-3p inhibits the degenerate process of chondrogenic differentiation through modification of DNA methylation

Abstract

The aim of this work was to determine whether miR-455-3p regulates DNA methylation during chondrogenic differentiation of hMSCs. The expression of miR-455-3p and de novo methyltransferase DNMT3A was assessed in micromass culture of hBMSCs, which induced chondrogenic differentiation in vitro, and in E16.5 mice in vivo. A luciferase reporter assay was used to confirm whether miR-455-3p directly targets DNMT3A by interaction with the 3′-UTR. Using an Illumina Infinium Methylation EPIC microarray, genome-wide DNA methylation of hBMSCs with or without overexpressed miR-455-3p was examined for 28 days during induced chondrogenic differentiation. Here, we showed that miR-455-3p was more expressed during the middle stage of hBMSC chondrogenic differentiation, and less expressed in the late stage. DNMT3A was less expressed in the middle stage and more expressed in the late stage, and was also more expressed in the palms of miR-455-3p deletion mice compared to those of wild-type mice. The luciferase reporter assay demonstrated that miR-455-3p directly targets DNMT3A 3′-UTR. miR-455-3p overexpression inhibits the degenerate process during chondrogenic differentiation, while deletion of miR-455-3p in mice accelerated cartilage degeneration. Genome-wide DNA methylation analysis showed miR-455-3p overexpression regulates DNA methylation of cartilage-specific genes. GO analysis revealed PI3K-Akt signaling pathway was most hypomethylated. Our data show that miR-455-3p can regulate hMSC chondrogenic differentiation by affecting DNA methylation. Overexpression of miR-455-3p and DNA methylation inhibitors can thus potentially be utilized to optimize chondrogenic differentiation.

Substantive molecular and histological changes within the meniscus with tears

“…Recent advances in epigenetic research have shed light on the importance of microRNA (miRNA) in the regulation of gene expression at multiple levels related to the pathogenesis of OA [18]. In a previous study, we reported significant up-regulation of the miRNAs, miR-381-3p, miR-455-3p, miR-193b-3p, and miR-92a-3p during differentiation of human mesenchymal stem cells into chondrocytes, and provided evidence that these four miRNAs may regulate early chondrogenesis and cartilage degeneration [19][20][21][22][23][24]. However, the expression profile of the miRNAs in the menisci with tears is unknown.…”

Long Non-coding RNA HOTTIP Promotes CCL3 Expression and Induces Cartilage Degradation by Sponging miR-455-3p

“…MiRNAs (∼22-nt) are robust regulators of post-transcriptional gene expression with a high affinity for the 3′ untranslated region, ultimately carrying out their function through recruitment of protein complexes that block ribosomal translation and contribute to degradation of the poly-adenylated tail (Bartel, 2004). Previously, we reported a 2.973-fold upregulation of miR-455-3p upon differentiation of human adipose-derived stem cells into chondrocytes (Zhang et al, 2012) and provided evidence that miR-455-3p is an important regulator of chondrogenesis and cartilage degeneration via the inhibition of RUNX2/HDAC2/HDAC8/DNMT3A (Zhang et al, 2015; Chen et al, 2016; Sun H. et al, 2018). Although studies of miRNAs have dominated the field of RNA biology in recent years, accumulating evidence shows that lncRNAs (>200-nt) are central regulators of biological processes, including those involved in the pathology of OA (Liu et al, 2014; Xing et al, 2014; Pearson et al, 2016); they regulate gene expression at both the transcriptional and post-transcriptional levels (Yang et al, 2014).…”

DNA methylation of noncoding RNAs: new insights into osteogenesis and common bone diseases

“…A new regulatory mechanism has been identified that ncRNAs, such as lncRNAs and miR-NAs, can be potential triggers in the decision of cell fates by methylated modification. For example, lncRNA Plnc1 was reported to mediate the differentiation of bone marrow stromal cells (BMSCs) into adipocytes by DNA methylation [32], and miRNA-455-3p changed the methylation status of chondrogenic-specific genes during the differentiation of human bone marrow mesenchymal stem cells (hBMSCs) towards chondrocytes [33], which would be discussed in more detail in later sections.…”

“…It was reported that miR-455-3p directly targeted the 3′-UTR of DNMT3A and regulated the process of chondrogenic differentiation in hBMSCs by altering the methylation levels of genes associated with cartilage development. Most of these genes, including FOXO3A, SMAD3, COL11A1, and SOX6, were hypomethylated and involved in the P13K-Akt signaling pathway [33], revealing the hypomethylated signaling pathway was a crucial regulator of chondrogenic differentiation.…”