GATA4 inhibits odontoblastic differentiation of dental pulp stem cells through targeting IGFBP3

Yan Zhang; Weiwei Qiao; Yaoting Ji; Liuyan Meng

doi:10.1016/j.archoralbio.2023.105756

GATA4 inhibits odontoblastic differentiation of dental pulp stem cells through targeting IGFBP3

Arch Oral Biol. 2023 Oct:154:105756. doi: 10.1016/j.archoralbio.2023.105756. Epub 2023 Jun 30.

Authors

Yan Zhang¹, Weiwei Qiao¹, Yaoting Ji², Liuyan Meng³

Affiliations

¹ State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan, China.
² State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan, China. Electronic address: yaotingji@whu.edu.cn.
³ State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan, China. Electronic address: mengliuyan@whu.edu.cn.

PMID: 37451139
DOI: 10.1016/j.archoralbio.2023.105756

Abstract

Objective: The odontogenic differentiation of human dental pulp stem cells (HDPSCs) is associated with reparative dentinogenesis. Transcription factor GATA binding protein 4 (GATA4) is proved to be essential for osteoblast differentiation and bone remodeling. This study clarified the function of GATA4 in HDPSCs odontoblast differentiation.

Methods: The change in GATA4 expression during reparative dentin formation was detected by immunohistochemistry staining. The expression of GATA4 during HDPSCs odontoblastic differentiation was detected by western blot and quantitative polymerase chain reaction. The effect of GATA4 on odontoblast differentiation was investigated following overexpression lentivirus transfection. RNA sequencing, dual luciferase assay and chromatin immunoprecipitation (CHIP) were conducted to verify downstream targets of GATA4. GATA4 overexpression lentivirus and small interference RNA targeting IGFBP3 were co-transfected to investigate the regulatory mechanism of GATA4.

Results: Upregulated GATA4 was observed during reparative dentin formation in vivo and the odontoblastic differentiation of HDPSCs in vitro. GATA4 overexpression suppressed the odontoblastic potential of HDPSCs, demonstrated by decreased alkaline phosphatase activity (p < 0.0001), mineralized nodules formation (p < 0.01), and odonto/osteogenic differentiation markers levels (p < 0.05). RNA sequencing revealed IGFBP3 was a potential target of GATA4. CHIP and dual luciferase assays identified GATA4 could activate IGFBP3 transcription. Additionally, IGFBP3 knockdown recovered the odontoblastic differentiation defect caused by GATA4 overexpression (p < 0.05).

Conclusions: GATA4 inhibited odontoblastic differentiation of HDPSCs via activating the transcriptional activity of IGFBP3, identifying its promising role in regulating HDPSCs odontoblast differentiation and reparative dentinogenesis.

Keywords: GATA4; Human dental pulp stem cells; IGFBP3; Odontoblastic differentiation; Reparative dentin; Transcription factor.

MeSH terms

Cell Differentiation / physiology
Cells, Cultured
Dental Pulp*
GATA4 Transcription Factor / genetics
GATA4 Transcription Factor / metabolism
Humans
Insulin-Like Growth Factor Binding Protein 3 / genetics
Insulin-Like Growth Factor Binding Protein 3 / metabolism
Odontoblasts
Osteogenesis*
Stem Cells

Substances

GATA4 protein, human
GATA4 Transcription Factor
IGFBP3 protein, human
Insulin-Like Growth Factor Binding Protein 3