GREM1 Negatively Regulates Osteo-/Dentinogenic Differentiation of Dental Pulp Stem Cells via Association with YWHAH.

Shu Diao, Xiao Han, Wei Long Ye, Chen Zhang, Dong Mei Yang, Zhi Peng Fan, Song Lin Wang
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Abstract

Objective: To investigate the biological regulatory function of Gremlin1 (GREM1) and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein eta (YWHAH) in dental pulp stem cells (DPSCs), and determine the underlying molecular mechanism involved.

Methods: Alkaline phosphatase (ALP) activity, alizarin red staining, scratch migration assays and in vitro and in vivo osteo-/dentinogenic marker detection of bone-like tissue generation in nude mice were used to assess osteo-/dentinogenic differentiation. Coimmunoprecipitation and polypeptide microarray assays were employed to detect the molecular mechanisms involved.

Results: The data revealed that knockdown of GREM1 promoted ALP activity, mineralisation in vitro and the expression of osteo-/dentinogenic differentiation markers and enhanced osteo-/ dentinogenesis of DPSCs in vivo. GREM1 bound to YWHAH in DPSCs, and the binding site was also identified. Knockdown of YWHAH suppressed the osteo-/dentinogenesis of DPSCs in vitro, and overexpression of YWHAH promoted the osteo-/dentinogenesis of DPSCs in vitro and in vivo.

Conclusion: Taken together, the findings highlight the critical roles of GREM1-YWHAH in the osteo-/dentinogenesis of DPSCs.

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GREM1 通过与 YWHAH 的关联负性调控牙髓干细胞的骨质/牙本质分化
目的研究牙髓干细胞(DPSCs)中Gremlin1(GREM1)和酪氨酸3-单氧化酶/色氨酸5-单氧化酶活化蛋白eta(YWHAH)的生物调控功能,并确定其潜在的分子机制:方法:采用碱性磷酸酶(ALP)活性、茜素红染色、划痕迁移试验和裸鼠体内外骨/牙源性标志物检测来评估骨/牙源性分化。采用免疫共沉淀和多肽芯片检测相关分子机制:数据显示,敲除 GREM1 可促进 ALP 活性、体外矿化和成骨/成牙分化标记物的表达,并增强体内 DPSCs 的成骨/成牙作用。GREM1 与 DPSCs 中的 YWHAH 结合,并确定了结合位点。体外敲除YWHAH抑制了DPSCs的骨/牙质生成,而过表达YWHAH促进了DPSCs在体外和体内的骨/牙质生成:综上所述,研究结果凸显了GREM1-YWHAH在DPSCs骨/牙生成过程中的关键作用。
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