A NOTCH1/LSD1/BMP2 co-regulatory network mediated by miR-137 negatively regulates osteogenesis of human adipose-derived stem cells

Abstract

Background MicroRNAs have been recognized as critical regulators for the osteoblastic lineage differentiation of human adipose-derived stem cells (hASCs). Previously, we have displayed that silencing of miR-137 enhances the osteoblastic differentiation potential of hASCs partly through the coordination of lysine-specific histone demethylase 1 (LSD1), bone morphogenetic protein 2 (BMP2), and mothers against decapentaplegic homolog 4 (SMAD4). However, still numerous molecules involved in the osteogenic regulation of miR-137 remain unknown. This study aimed to further elucidate the epigenetic mechanisms of miR-137 on the osteogenic differentiation of hASCs. Methods Dual-luciferase reporter assay was performed to validate the binding to the 3′ untranslated region (3′ UTR) of NOTCH1 by miR-137. To further identify the role of NOTCH1 in miR-137-modulated osteogenesis, tangeretin (an inhibitor of NOTCH1) was applied to treat hASCs which were transfected with miR-137 knockdown lentiviruses, then together with negative control (NC), miR-137 overexpression and miR-137 knockdown groups, the osteogenic capacity and possible downstream signals were examined. Interrelationships between signaling pathways of NOTCH1-hairy and enhancer of split 1 (HES1), LSD1 and BMP2-SMADs were thoroughly investigated with separate knockdown of NOTCH1, LSD1, BMP2, and HES1. Results We confirmed that miR-137 directly targeted the 3′ UTR of NOTCH1 while positively regulated HES1. Tangeretin reversed the effects of miR-137 knockdown on osteogenic promotion and downstream genes expression. After knocking down NOTCH1 or BMP2 individually, we found that these two signals formed a positive feedback loop as well as activated LSD1 and HES1. In addition, LSD1 knockdown induced NOTCH1 expression while suppressed HES1. Conclusions Collectively, we proposed a NOTCH1/LSD1/BMP2 co-regulatory signaling network to elucidate the modulation of miR-137 on the osteoblastic differentiation of hASCs, thus providing mechanism-based rationale for miRNA-targeted therapy of bone defect.

Microfibrillar-associated protein 5 regulates osteogenic differentiation by modulating the Wnt/β-catenin and AMPK signaling pathways

“…Of note, MFAP5 activated Notch 1 signaling in certain tumors, promoting tumor invasion and migration (Chen et al 2020 ; Li et al 2019 ). Notch 1 signaling is also involved in osteogenesis (Díaz-Tocados et al 2017 ; Fan et al 2016 ; Fan et al 2021 ). However, the Notch 1 levels of the MFAP5 knockdown and control groups did not differ significantly (see Additional file 2 ).…”

It takes two to tango: Widening our understanding of the onset of schizophrenia from a neuro-angiogenic perspective

“…It has been reported that Notch1 is a target of miR-137, which acts as a regulator of Notch signaling in neurons. Consistent observations in human adipose cells have shown that miR-137 binds directly to the NOTCH1 3′UTR region and ultimately positively regulates HES1 (Shi et al, 2017;Fan et al, 2021). Recent evidence provides insight into the role of miR-137 in response to oxidative damage and neuroprotection with a proposed neuroprotector and anti-inflammatory role against chemical agents and artery occlusion (Y.…”

MiR-137-mediated negative relationship between LGR4 and RANKL modulated osteogenic differentiation of human adipose-derived mesenchymal stem cells

“…The plasmids of negative control (NC), miR‐137 overexpression (miR‐137), miR‐137 knockdown (anti‐miR‐137), LGR4 shRNA (anti- LGR4 ) and RANKL shRNA (anti- RANKL ) were separately employed to generate the corresponding lentiviruses. The hASCs were transfected by respective lentiviruses as previously described ( Ma et al , 2020 ; Fan et al , 2021 ). After being immersed in the viral supernatant (multiplicity of infection = 100) with 5 mg/mL polybrene (Sigma-Aldrich) for 24 h, the cells were then selected by 1 μg/ml puromycin (Sigma-Aldrich) and cultured in fresh PM.…”

“…As an ideal stem cell source, human adipose-derived mesenchymal stem cells (hASCs) are widely used in bone tissue engineering due to their accessibility, multipotency and low immunogenicity ( Dinescu et al , 2021 ). MicroRNAs (miRNAs), a class of small non-coding RNAs ranging from 19-25 nucleotides in size, regulate protein-coding genes post-transcriptionally and play crucial roles in various biological activities, including the proliferation and osteogenic differentiation of hASCs ( Ranganathan and Sivasankar, 2014 ; Fan et al , 2021 ). It has been demonstrated that miR-137 acts as a key regulator in various kinds of malignancies ( Li et al , 2017 ; Liu et al , 2017 ; Qin et al , 2017 ; Luo et al , 2018 ; Zhang et al , 2018 ) and neural development ( Silber et al , 2008 ; Szulwach et al , 2010 ; Tarantino et al , 2010 ; Sun et al , 2011 ; Jiang et al ,2013 ).…”

“…It has been demonstrated that miR-137 acts as a key regulator in various kinds of malignancies ( Li et al , 2017 ; Liu et al , 2017 ; Qin et al , 2017 ; Luo et al , 2018 ; Zhang et al , 2018 ) and neural development ( Silber et al , 2008 ; Szulwach et al , 2010 ; Tarantino et al , 2010 ; Sun et al , 2011 ; Jiang et al ,2013 ). But up to present, only a few studies investigated the effects of miR-137 on osteogenesis ( Zheng et al , 2019 ; Kong et al , 2020 ; Ma et al , 2020 ; Yu et al , 2020 ; Fan et al , 2021 ), and the mechanisms of osteogenic modulation by miR-137 are still not fully understood, especially considering the diverse biological properties of different cell types.…”