Regulation of the IGFBP-5 and MMP-13 genes by the microRNAs miR-140 and miR-27a in human osteoarthritic chondrocytes

Abstract

BackgroundMMP-13 and IGFBP-5 are important factors involved in osteoarthritis (OA). We investigated whether two highly predicted microRNAs (miRNAs), miR-140 and miR-27a, regulate these two genes in human OA chondrocytes.MethodsGene expression was determined by real-time PCR. The effect of each miRNA on IGFBP-5 and MMP-13 expression/production was evaluated by transiently transfecting their precursors (pre-miRNAs) and inhibitors (anti-miRNAs) into human OA chondrocytes. Modulation of IGFBP-5, miR-140 and miR-27a expression was determined upon treatment of OA chondrocytes with cytokines and growth factors.ResultsIGFBP-5 was expressed in human chondrocytes with its level significantly lower (p < 0.04) in OA. Five computational algorithms identified miR-140 and miR-27a as possible regulators of MMP-13 and IGFBP-5 expression. Data showed that both miRNAs were expressed in chondrocytes. There was a significant reduction (77%, p < 0.01) in miR-140 expression in OA compared to the normal chondrocytes, whereas miR-27a expression was only slightly decreased (23%). Transfection with pre-miR-140 significantly decreased (p = 0.0002) and with anti-miR-140 significantly increased (p = 0.05) IGFBP-5 expression at 24 hours, while pre-miR-27a did not affect either MMP-13 or IGFBP-5. Treatment with anti-miR-27a, but not with anti-miR-140, significantly increased the expression of both MMP-13 (p < 0.05) and IGFBP-5 (p < 0.01) after 72 hours of incubation. MMP-13 and IGFBP-5 protein production followed the same pattern as their expression profile. These data suggest that IGFBP-5 is a direct target of miR-140, whereas miR-27a down-regulates, likely indirectly, both MMP-13 and IGFBP-5.ConclusionThis study is the first to show the regulation of these miRNAs in human OA chondrocytes. Their effect on two genes involved in OA pathophysiology adds another level of complexity to gene regulation, which could open up novel avenues in OA therapeutic strategies.

MicroRNA Levels as Prognostic Markers for the Differentiation Potential of Human Mesenchymal Stromal Cell Donors

“…We next extended our analysis by analysis of a selected panel of miRNAs previously implemented in chondrogenic differentiation, including miR-210, miR-630, and miR-140 [40][41][42][43]. We confirmed positive regulation of miR-210 and miR-630 within 7 days after chondrogenic induction, with miR-630 being slightly higher expressed in good donors.…”

MicroRNAs and post-transcriptional regulation of skeletal development

“…Besides its known role in chondrocyte differentiation, miR-140 also plays a central role in OA (Araldi & Schipani 2010). In vivo, miR-140 and miR-146a target ADAMTS5 (Miyaki et al 2009, Tardif et al 2009, while miR-125b targets ADAMTS4 (Matsukawa et al 2013). Moreover, miR-101 directly inhibits the expression of collagen type II and aggrecan genes through the downregulation of SOX9 expression (Dai et al 2012).…”

“…Numerous miRNAs have been shown to be regulated by IL1b action (Miyaki et al 2009, Dai et al 2012, Matsukawa et al 2013. MMP13 is regulated directly by miR-27b and miR-127-5p (Akhtar et al 2010 or indirectly by miR-27a, miR-9, miR-488, or miR-22 (Iliopoulos et al 2008, Jones et al 2009, Tardif et al 2009, Song et al 2013b, among others.…”

MicroRNA‐199a‐3p, microRNA‐193b, and microRNA‐320c are correlated to aging and regulate human cartilage metabolism

“…2 Abnormalities in cartilage development have been observed in Dicer gene dysfunction models, and miRNA is known to play a key role in cartilage differentiation. 3 miRNA plays a key role in differentiation and growth in organs and tissues, 2 and miRNAs expressed specifically in cartilage [4][5][6][7][8] have been reported. Till date, miR21, 4 miR22, 9 miR27a, 6 miR140, 10 and miR146 8 have been reported as miRNAs that are associated with cartilage metabolism.…”