Overexpression of miR-1306-5p, miR-3195, and miR-3914 Inhibits Ameloblast Differentiation through Suppression of Genes Associated with Human Amelogenesis Imperfecta

Abstract

Amelogenesis imperfecta is a congenital form of enamel hypoplasia. Although a number of genetic mutations have been reported in humans, the regulatory network of these genes remains mostly unclear. To identify signatures of biological pathways in amelogenesis imperfecta, we conducted bioinformatic analyses on genes associated with the condition in humans. Through an extensive search of the main biomedical databases, we found 56 genes in which mutations and/or association/linkage were reported in individuals with amelogenesis imperfecta. These candidate genes were further grouped by function, pathway, protein–protein interaction, and tissue-specific expression patterns using various bioinformatic tools. The bioinformatic analyses highlighted a group of genes essential for extracellular matrix formation. Furthermore, advanced bioinformatic analyses for microRNAs (miRNAs), which are short non-coding RNAs that suppress target genes at the post-transcriptional level, predicted 37 candidates that may be involved in amelogenesis imperfecta. To validate the miRNA–gene regulation association, we analyzed the target gene expression of the top seven candidate miRNAs: miR-3195, miR-382-5p, miR-1306-5p, miR-4683, miR-6716-3p, miR-3914, and miR-3935. Among them, miR-1306-5p, miR-3195, and miR-3914 were confirmed to regulate ameloblast differentiation through the regulation of genes associated with amelogenesis imperfecta in AM-1 cells, a human ameloblastoma cell line. Taken together, our study suggests a potential role for miRNAs in amelogenesis imperfecta.

MicroRNA-1306-5p Regulates the METTL14-Guided m6A Methylation to Repress Acute Myeloid Leukemia

“…Several miRNAs are also found to play crucial roles in AML by regulating various kinds of biological processes, such as miRNA-485-5p, miR-126, and miR-204 [12][13][14]. In addition, as reported previously, miR-1306-5p is involved in the progression of many diseases, such as amelogenesis imperfecta, cerebral ischemia/reperfusion injury, and sepsis [15][16][17]. However, the role of miR-1306-5p in AML is obscure.…”

“…miR-1306-5p was found to decrease cerebral ischemia/reperfusion injury in vitro by targeting BIK [ 16 ]. It was also confirmed to regulate ameloblast differentiation by regulating genes related to amelogenesis imperfecta [ 15 ] and inhibit the malignant behavior of osteosarcoma cells [ 32 ]. However, the specific biological roles of miR-1306-5p and its indirect interactions and regulation of m6A in AML remain poorly understood.…”

Epigenetics in developmental defects of enamel: A scoping review

“…MiR-exon4 can promote the expression of Runx2 and also results in the increase of the differentiation of ameloblasts (Le et al, 2016). By injecting miRNA mimics into human ameloblastoma AM-1 cells and then inducing amelogenesis differentiation, miR-1306-5, miR-3195 and miR-3914 were proved to regulate the genes related to enamel hypoplasia such as AMELX and KLK4, and thus affected amelogenesis especially in the secretion and mineralization processes (Yoshioka et al, 2021). Additionally, miR-16-5p and miR-27b-3p were confirmed to act similarly in another vitro experiment (Suzuki et al, 2022).…”

“…Through bioinformatic analysis, the genes related to enamel hypoplasia were searched by HirokiYoshioka et al and in in vitro experiments, miR-1306-5, miR-3195, and miR-3914 were proved to regulate the genes related to enamel hypoplasia and thus affected amelogenesis. By injecting miRNA mimics into human ameloblastoma AM-1 cells, they found that miR-1306-5p inhibited the expression of AMTN and KLK4 genes; miR-3195 inhibited AMELX, KLK4, MMP20; miR-3914 inhibited AMELX, KLK4(Yoshioka et al, 2021).Through a similar method, Akiko Suzuki et al discovered that miR-16-5p inhibited AMELX, miR-27b-3p inhibited AMELX and ENAM in mHAT9d cells, a mouse dental epithelial cell line(Suzuki et al, 2022).Cleidocranial dysplasia (CCD) is a congenital disease mainly caused by the Runx2 gene mutation on chromosome 6p21. Enamel hypoplasia is a common clinical manifestation in CCD patients.Runx2 may regulate the expression of miRNAs involved in bone and tooth development.…”