MicroRNA-27b targets CBFB to inhibit differentiation of human bone marrow mesenchymal stem cells into hypertrophic chondrocytes

Abstract

Background Human bone marrow-derived mesenchymal stem cells (hBMSCs) have chondrocyte differentiation potential and are considered to be a cell source for cell-transplantation-mediated repair of cartilage defects, including those associated with osteoarthritis (OA). However, chondrocyte hypertrophic differentiation is a major obstacle for the application of hBMSCs in articular cartilage defect treatment. We have previously shown that microRNA-27b (miR-27b) inhibits hypertrophy of chondrocytes from rat knee cartilage. In this study, we investigated the role of miR-27b in chondrocyte hypertrophic differentiation of hBMSCs. Methods Chondrogenic marker and microRNA expression in hBMSC chondrogenic pellets were evaluated using RT-qPCR and immunohistochemistry. The hBMSCs were transfected with miR-27b before inducing differentiation. Gene and protein expression levels were analyzed using RT-qPCR and western blot. Coimmunoprecipitation was used to confirm interaction between CBFB and RUNX2. Luciferase reporter assays were used to demonstrate that CBFB is a miR-27b target. Chondrogenic differentiation was evaluated in hBMSCs treated with shRNA targeting CBFB. Chondrogenic hBMSC pellets overexpressing miR-27b were implanted into cartilage lesions in model rats; therapeutic effects were assessed based on histology and immunohistochemistry. Results The hBMSCs showed typical MSC differentiation potentials. During chondrogenic differentiation, collagen 2 and 10 (COL2 and COL10), SOX9, and RUNX2 expression was upregulated. Expression of miR-140, miR-143, and miR-181a increased over time, whereas miR-27b and miR-221 were downregulated. Cartilage derived from hBMSC and overexpressing miR-27b exhibited higher expression of COL2 and SOX9, but lower expression of COL10, RUNX2, and CBFB than did the control cartilage. CBFB and RUNX2 formed a complex, and CBFB was identified as a novel miR-27b target. CBFB knockdown by shRNA during hBMSC chondrogenic differentiation led to significantly increased COL2 and SOX9 expression and decreased COL10 expression. Finally, miR-27b-overexpressing hBMSC chondrogenic pellets had better hyaline cartilage morphology and reduced expression of hypertrophic markers and tend to increase repair efficacy in vivo. Conclusion MiR-27b plays an important role in preventing hypertrophic chondrogenesis of hBMSCs by targeting CBFB and is essential for maintaining a hyaline cartilage state. This study provides new insights into the mechanism of hBMSC chondrocyte differentiation and will aid in the development of strategies for treating cartilage injury based on hBMSC transplantation.

miR-23a/b clusters are not essential for the pathogenesis of osteoarthritis in mouse aging and post-traumatic models

“…The expression of miR-23a, miR-24 and miR-27a was significantly decreased in OA chondrocytes compared with normal chondrocytes consistent with prior findings about miR-140 ( Figure 1C ). Thus, previous inconsistent reports ( Akhtar et al, 2010 ; Philipot et al, 2014 ; Hu et al, 2017 ; Zhou et al, 2017 ; Xu et al, 2018 ; Lv et al, 2020 ; Xu et al, 2021 ) and our results prompted us to examine the role of miR-23a/b clusters in cartilage development and OA pathogenesis using miR-23a/b clusters-deficient mice. We expected that 23a/b clusters-deficient mice would exhibit abnormal skeletal development and altered onset or severity of OA.…”

“…In the present study, we determined the role of miR-23a/b clusters, which are highly expressed in cartilage, in the pathogenesis of OA using two OA models in miR-23a/b clusters KO mice. Among miR-23a/b clusters, miR-23a, miR-23b and miR-27b have been suggested to have a role in Frontiers in Cell and Developmental Biology frontiersin.org maintaining articular cartilage state and anti-inflammation by in vitro studies (Akhtar et al, 2010;Hu et al, 2017;Zhou et al, 2017;Xu et al, 2018;Lv et al, 2020;Zhou et al, 2021). On the other hand, several reports suggest that miR-23a and miR-23b contribute to OA progression (Kang et al, 2016;Guo et al, 2018;Zhao et al, 2019).…”

“…Each individual miRNA (miR-23a, miR-23b, miR-24, miR-27a, and miR-27b) is finally processed from each cluster for the display of their function as matured miRNAs. Previous reports indicate that among miR-23a/b clusters, miR-23a, miR-24, and miR-27b have a role in maintaining articular cartilage integrity ( Akhtar et al, 2010 ; Philipot et al, 2014 ; Hu et al, 2017 ; Zhou et al, 2017 ; Xu et al, 2018 ; Lv et al, 2020 ; Xu et al, 2021 ). MiR-27, similar to miR-140, is downregulated in patients with OA ( Akhtar et al, 2010 ; Zhou et al, 2017 ; Xu et al, 2018 ).…”

MiR-27b-3p Targeting BDNF Inhibits TrkB / CREB Signaling Pathway and Improves IL-1 β Induced Chondrocytic Inflammation

“…The role of IL-1β+BDNF was inhibited miR-27b-3p mimics. Our data showed miR-27b-3p could target BDNF to inhibit TrkB/CREB pathway activation and chondrocyte apoptosis (36).…”

Integrated Analysis of the Transcriptome Profile Reveals the Potential Roles Played by Long Noncoding RNAs in Immunotherapy for Sarcoma

“…Recently, immunotherapy has been approved for different cancers, such as melanoma, prostate cancer, lymphoma, renal cell carcinoma, and breast cancer (39)(40)(41)(42)(43)(44)(45)(46)(47)(48)(49)(50)(51)(52)(53). The most successful of these strategies involve immune checkpoint inhibitors.…”