ETV2 promotes osteogenic differentiation of human dental pulp stem cells through the ERK/MAPK and PI3K-Akt signaling pathways

Abstract

Background The repair of cranio-maxillofacial bone defects remains a formidable clinical challenge. The Ets variant 2 (ETV2) transcription factor, which belongs to the E26 transformation-specific (ETS) family, has been reported to play a key role in neovascularization. However, the role of ETV2 in the osteogenesis of human dental pulp stem cells (hDPSCs) remains unexplored. Methods Transgenic overexpression of ETV2 was achieved using a lentiviral vector, based on a Dox-inducible system. The effects of Dox-induced overexpression of ETV2 on the osteogenesis of hDPSCs were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR), western blot, immunofluorescence staining, alkaline phosphatase (ALP) staining, and Alizarin Red S (ARS) staining. Additionally, RNA-sequencing (RNA-Seq) analysis was performed to analyze the underlying mechanisms of ETV2-induced osteogenesis. Additionally, the role of ETV2 overexpression in bone formation in vivo was validated by animal studies with a rat calvarial defect model and a nude mice model. Results Our results demonstrated that ETV2 overexpression significantly upregulated the mRNA and protein expression levels of osteogenic markers, markedly enhanced ALP activity, and promoted matrix mineralization of hDPSCs. Moreover, the results of RNA-Seq analysis and western blot showed that the ERK/MAPK and PI3K-Akt signaling pathways were activated upon transgenic overexpression of ETV2. The enhanced osteogenic differentiation of hDPSCs due to ETV2 overexpression was partially reversed by treatment with inhibitors of ERK/MAPK or PI3K-AKT signaling. Furthermore, the results of in vivo studies demonstrated that ETV2 overexpression improved bone healing in a rat calvarial defect model and increased ectopic bone formation in nude mice. Conclusions Collectively, our results indicated that ETV2 overexpression exerted positive effects on the osteogenesis of hDPSCs, at least partially via the ERK/MAPK and PI3K/AKT signaling pathways.

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“…PI3K-AKT is an important intracellular pathway that can regulate cell survival, proliferation, protein synthesis, and other cell activities. The importance of PI3K-AKT pathway on osteogenesis has been proved by some studies ( 59 , 60 ), and the expression of PI3K and AKT protein was up-regulated under ES as proved by Western blot analysis ( Fig. 5L ).…”

Multifunctional Prosthesis Surface: Modification of Titanium with Cinnamaldehyde‐Loaded Hierarchical Titanium Dioxide Nanotubes

“…Simultaneously, we observed effective activation of the MAPK signaling pathway, which is also a classical pathway involved in bone formation. [ 68,69 ] This was manifested by a significant increase in the expression levels of phosphorylated ERK and JNK in the NT/CIN group compared to those in the PT group (Figure 6B,E, Supporting Information). In conclusion, these findings highlight the efficient promotion of osteogenic capability through NT/CIN surface modification and shed light on the potential cellular signaling pathways involving Wnt/β‐catenin and MAPK (Figure 6C).…”

ETV2 regulating PHD2-HIF-1α axis controls metabolism reprogramming promotes vascularized bone regeneration

“…Our previous research has established that ETV2 promotes ERK1/2 phosphorylation, which may facilitate the formation of the HIF-1α and HIF-1β heterodimer and translocate into the nucleus for transcriptional activity [ 15 , 28 ]. Through the separation of cytoplasmic and nucleoprotein, we observed that ETV2 substantially increased intracellular phosphorylated ERK1/2 (pERK1/2) and intranuclear HIF-1α ( Fig.…”