Gene expression regulatory network and molecular mechanism involved in odontogenic differentiation of human dental pulp stem cells-derived induced pluripotent stem cells

doi:10.12307/2026.421

Abstract

Abstract: BACKGROUND: Human dental pulp stem cell-derived induced pluripotent stem cells possess self-renewal and multidirectional differentiation potential, showing great promise in promoting dentin regeneration. However, the mechanism of their odontogenic differentiation remains unclear.
OBJECTIVE: To analyze the hub genes and potential molecular mechanisms during the odontogenic differentiation of human dental pulp stem cell-derived induced pluripotent stem cells.
METHODS: Total RNA of human dental pulp stem cell-derived induced pluripotent stem cells and their differentiated odontogenic cells were extracted for RNA-sequencing to analyze the transcriptome gene expression profile. Firstly, the differentially expressed mRNAs, miRNAs, long non-coding RNAs, and circular RNAs were screened to identify key genes. Gene Set Enrichment Analysis was performed to analyze the key signaling pathways. Subsequently, a competitive endogenous RNA (ceRNA) network and a transcription factor-mRNA-miRNA network were constructed to predict and reveal the regulatory mechanisms of key genes. Finally, real-time quantitative PCR was used to verify the expression of key genes.
RESULTS AND CONCLUSION: Six hub genes were identified during the odontogenic differentiation of human dental pulp stem cell-derived induced pluripotent stem cells: Smad family member 3, signal transducer and activator of transcription 3, fibronectin 1, estrogen receptor 1, epidermal growth factor receptor, and enhancer of zeste homolog 2. The most enriched pathways included Wnt and transforming growth factor β signaling pathways. CeRNA network analysis revealed that hsa-miR-302, hsa-miR-519, and hsa-miR-520 were the main regulatory factors of estrogen receptor 1, with multiple interaction pairs. Transcription factor-mRNA network analysis showed that interferon regulatory factor 3 was a regulatory factor for all six hub genes, while enhancer of zeste homolog 2 was regulated by four transcription factors simultaneously. Real-time quantitative PCR results showed that the expression of fibronectin 1 and enhancer of zeste homolog 2 was significantly increased in odontogenic cells differentiated from dental pulp stem cell-derived induced pluripotent stem cells, while the expression of signal transducer and activator of transcription 3 was significantly decreased.

Key words: human dental pulp stem cells, induced pluripotent stem cells, odontogenic differentiation, RNA sequencing, key genes, differentially expressed genes, gene regulatory network, competitive endogenous RNA network

CLC Number:

Rong Qiong, Guo Yu, Wang Tingting, Tan Xiaobing. Gene expression regulatory network and molecular mechanism involved in odontogenic differentiation of human dental pulp stem cells-derived induced pluripotent stem cells[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(31): 8122-8134.

Figures/Tables 8

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