Mechanisms of miR‑128‑3p in inhibiting osteoblast differentiation from bone marrow‑derived mesenchymal stromal cells

Abstract

The authors' previous study demonstrated that mir-128 may exert an inhibitory effect on the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BM-MScs), but its downstream mechanisms remain to be elucidated. The aim of the present study was to investigate the microRNA (miRNA/miR) and mRNA profiles of differentiated and undifferentiated BM-MScs and explore new downstream targets for mir-128. The sequencing datasets of GSe107279 (mirna) and GSe112318 (mrna) were downloaded from the Gene expression omnibus database. The differentially expressed mirnas (deMs) and genes (deGs) were identified using the DESeq2 method. The target genes of deMs were predicted by the mirwalk 2.0 database. The hub target genes of mir-128 were screened by constructing the protein-protein interaction (PPi) network and module analysis. The expression levels of mir-128 and crucial target genes were validated by reverse transcription-quantitative (rT-q) Pcr before or after transfection of mir-128 mimics to BM-MSCs. The miRNA expression profile analysis identified miR-128 as one of the significantly downregulated DEMs (total 338) in differentiated BM-MScs compared with the undifferentiated control. a total of 103 predicted target genes of mir-128-3p were overlapped with upregulated deGs. By calculating the topological properties of each protein in the PPi network, 6 upregulated genes (KIT, NTRK2, YWHAB, GAB1, AXIN1 and RUNX1; fold change was the highest for NTRK2) were considered to be hub genes. of these, 4 were enriched in module 4 (RUNX1, KIT, GAB1 and AXIN1; RUNX1 was particularly crucial as it can interact with the others), while one was enriched in module 7 (YWHAB). The expression levels of mir-128 and these 6 target genes during the osteogenic differentiation were experimentally confirmed by rT-qPcr. in addition, the expression levels of these 6 genes were significantly reversed after transfection of miR-128-3p mimics into rat BM-MScs compared with the mir-control group. These findings indicated that miR-128-3p may inhibit the osteoblast differentiation of BM-MScs by downregulation of these 6 genes, particularly RUNX1, YWHAB and NTRK2.

LncRNA USP2-AS1 facilitates the osteogenic differentiation of bone marrow mesenchymal stem cells by targeting KDM3A/ETS1/USP2 to activate the Wnt/β-catenin signaling pathway

“…Firstly, we applied hTFtarget ( http://bioinfo.life.hust.edu.cn/hTFtarget#!/ ) to predict putative transcription factors of USP2 in bone marrow tissues. Since the role of ETS1 [ 23 ;] RUNX1 [ 24 ;] CTCF [ 25 ;] E2F4 [ 26 ] and MYC [ 27 , 28 ] in osteogenic differentiation have been reported, we selected them as the research candidates ( Figure 3A ). Subsequently, we knocked down these transcription factors and detected the interference efficiency (Fig.…”

Identification of a global gene expression signature associated with the genetic risk of catastrophic fracture in iPSC-derived osteoblasts from Thoroughbred horses

“…osteoclasts) also contribute to fracture risk. Human iPSCs have successfully been differentiated into osteoclasts (Chen, 2020), but to date this not yet been reported for equine iPSCs and it would be of benefit to measure gene expression profiles in other cell types in the future. Similarly, we did not have access to bone tissue from fracture cases and controls and so were unable to confirm if these differences in expression also occur in vivo at either the gene or protein level.…”

“…For example, SOSTDC1 has been shown to be involved in bone metabolism, the maintenance of bone density and fracture repair (Tong et al, 2022, Collette et al, 2016). NTRK2 is involved in osteoblast differentiation of MSCs (Zhang et al, 2020) and binding of NTRK2 to its receptor results in the phosphorylation of ERK1/2 which drives the expression of the bone-promoting transcription factors RUNX2 and SP7 (Osterix) (Liu et al, 2018). EPHA3 is a member of the ephrin receptor family and ephrin signalling is involved in bone development and homeostasis (Arthur and Gronthos, 2021).…”

miR-452-3p inhibited osteoblast differentiation by targeting Smad4

“…Several miRNAs that participate in the differentiation of osteoblasts have been identified. For example, miR-497-5p (Zhao et al, 2020), microRNA-150-3p (Qiu et al, 2020), miR-708-5p (Wang et al, 2020), and microRNA-497-5p (Liu et al, 2020a) have been shown to promote this process, while miR-30 (Zhang et al, 2020a), miR-129-5p (Yin et al, 2020), miR-128-3p (Zhang et al, 2020b), and miR-17-5p have suppressive effects. In various cell types, miR-425-3p is differentially expressed (Li et al, 2020;Ma et al, 2019), but its function in osteogenesis has not been documented.…”