Analysis of -5p and -3p Strands of miR-145 and miR-140 During Mesenchymal Stem Cell Chondrogenic Differentiation

Abstract

The chondrogenic differentiation of mesenchymal stem cells (MSCs) is mediated by transcription factors and small noncoding RNAs such as microRNAs (miRNAs). Each miRNA is initially transcribed as a long transcript, which matures to produce -5p and -3p strands. It is widely believed that the mature and functional miRNA from any given pre-miRNA, usually the -5p strand, is functional, while the opposing -3p strand is degraded. However, recent cartilage literature started to show functional -3p strands for a few miRNAs. This study aimed at examining both -5p and -3p strands of two key miRNAs miR-140 and miR-145, known to be involved in the chondrogenic differentiation of MSCs. The level (copy number) of both -5p and -3p strands of miR-145 and miR-140 along the time line of MSC chondrogenic differentiation was determined by polymerase chain reaction. The gene expression profiles of several genes related to MSC chondrogenesis were compared with these miRNA profiles along the same timeline. While miR-145-3p is declining in step with miR-145-5p in pellet cultures during the process, the -3p strand is only 1-2% of the total miR-145 products. In contrast, the mature -3p and -5p products of miR-140 are found to increase with near-equal molar expression throughout chondrogenic differentiation. Numerous genes are expressed by cartilage progenitor cells during development. One such target gene, Sox9, is a regulatory target of the dominant miR-145-5p, consistent with the data. Further experimental validations are warranted to confirm that ACAN, FOXO1, and RUNX3 as direct targets of miR-145-5p in the context of MSC chondrogenesis. Similarly, TRSP1 and ACAN are worth further validation as direct targets of miR-145-3p. For miR-140, SOX4 shall be further validated as a direct target of miR-140-5p, while KLF4, PTHLH, and WNT5A can be validated as direct targets of miR-140-3p.

Long Non-Coding RNA (lncRNA) Small Nucleolar RNA Host Gene 15 (SNHG15) Alleviates Osteoarthritis Progression by Regulation of Extracellular Matrix Homeostasis

“…We predicted 164 miRNAs targeting KLF4 using the ENCORI database ( http://starbase.sysu.edu.cn/ ). Among them, 8 KLF4-binding miRNAs were verified in previous studies: hsa-miR-346 [ 21 ], hsa-miR-140-3p [ 22 ], hsa-miR-18a [ 23 ], hsa-miR-15a [ 24 ], hsa-miR-103a [ 25 ], hsa-miR-7 [ 26 ], hsa-miR-135 [ 27 ], and hsa-miR-29a [ 28 ]. Furthermore, targeted miRNAs of downregulated lncRNAs were predicted using the InCeDb database ( http://gyanxet-beta.com/lncedb/ ).…”

Irisin enhances chondrogenic differentiation of human mesenchymal stem cells via Rap1/PI3K/AKT axis.

“…Accumulated evidences have shown that post-transcriptional regulation by microRNAs (miRNAs) was involved in the osteogenic and chondrogenic differentiation of MSCs [27,41] . It was a practical approach to mediate the chondrogenic differentiation of MSCs by manipulating the expression of speci c miRNAs [42][43][44] . A variety of miRNAs participated in the processes of irisin on osteogenic differentiation of MSCs, browning of white adipocytes, and anti-in ammation [19,[45][46] .…”

The Strand Specific Regulation of miR-155-3p in Response to Lipopolysaccharide and Interleukin-10 Stimulation Requires FUBP1 Protein

“…KSRP processes and controls the expression of miRNAs (miR-145, miR-150, let-7a) [29,42,43] other than miR-155, and the proposed mechanism involves recruitment of proteins such as Drosha, DGCR8, and Dicer [29]. Interestingly, all the miRNAs controlled by KSRP have the 5p strand as the dominantly expressed strand (guide strand) and the 3p strand as the passenger strand [44][45][46]. Thus, KSRP may be participating in strand selection by bringing specific proteins into the Dicer complex that enhances the thermodynamic stability [47] of the 5p strand binding to the Dicer protein.…”