Abstract
Objective To investigate the function of NAT10 in mesenchymal stem cell (MSC) osteogenic differentiation and study the mechanism by which NAT10 affects MSC osteogenesis by mediating Gremlin 1 N4-acetylcytidine (ac4C) modification. Methods Osteogenic differentiation of MSCs was induced, and the osteogenic ability was evaluated with alizarin red S (ARS) and alkaline phosphatase (ALP) assays. The NAT10 expression level during MSC osteogenesis was measured by western blot (WB). MSCs were transfected with lentiviruses to inhibit (Sh-NAT10) or overexpress NAT10 (Over-NAT10), and the osteogenic differentiation ability was assessed by ARS, ALP, and osteogenic gene marker assays. β-Catenin, Akt, and Smad signaling pathway component activation levels were assessed, and the expression levels of key Smad signaling pathway molecules were determined by PCR and WB. The Gremlin 1 mRNA ac4C levels were analyzed using RIP-PCR, and the Gremlin 1 mRNA degradation rate was determined. Sh-Gremlin 1 was transfected to further investigate the role of NAT10 and Gremlin 1 in MSC osteogenesis. Results During MSC osteogenesis, NAT10 expression, ARS staining, and the ALP level gradually increased. Decreasing NAT10 expression inhibited, and increasing NAT10 expression promoted MSC osteogenic differentiation. NAT10 affected the BMP/Smad rather than the Akt and β-Catenin signaling pathway activation by regulating Gremlin 1 expression. The Gremlin 1 mRNA ac4C level was positively regulated by NAT10, which accelerated Gremlin 1 degradation. Sh-Gremlin 1 abolished the promotive effect of NAT10 on MSC osteogenic differentiation. Conclusion NAT10 positively regulated MSC osteogenic differentiation through accelerating the Gremlin 1 mRNA degradation by increasing its ac4C level. These results may provide new mechanistic insight into MSC osteogenesis and bone metabolism in vivo.
Highlights
Mesenchymal stem cells (MSCs), first characterized in 1976, are stem cells with multipotency in vivo [1]
Our results showed that N-Acetyltransferase 10 (NAT10) increased the ac4C modification of Gremlin 1 Messenger RNAs (mRNAs) and accelerated its degradation rate, which activated the bone morphogenetic protein (BMP)/Smad signaling pathway, positively regulating the osteogenic differentiation ability of MSCs
The protein level of NAT10 increased in MSCs undergoing osteogenic differentiation, and its expression pattern was similar to that demonstrated by the alizarin red S (ARS) and alkaline phosphatase (ALP) assays (Figure 1(e))
Summary
Mesenchymal stem cells (MSCs), first characterized in 1976, are stem cells with multipotency in vivo [1]. In addition to their strong regulatory ability as components of the immune system, MSCs possess the capacity for trilineage differentiation, namely, osteogenic differentiation, chondrogenic differentiation, and adipogenic differentiation [2, 3]. The functions of MSCs are controlled by various kinds of RNA modifications [6]. N4-acetylcytidine (ac4C) modification was found to be widely distributed on RNA in human cells [7]. Messenger RNAs (mRNAs) modified by ac4C exhibit differences in their stability and degradation rate, affecting the expression of genes and the subsequent functions of cells [8]. N-Acetyltransferase 10 (NAT10) is a critical RNA acetyltransferase mediating ac4C
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
