Mechanism of HDAC3-Targeted Regulation of GATA-2 Affecting Osteogenic Differentiation of Dental Pulp Stem Cells

Qian OUYANG; Yanfei HUANG; Lingyun YIN; Lingpeng ZHANG; Jing LIANG; Hongbin YU

Article Contents

Article Navigation > Journal of Kunming Medical University > 2025 > Accepted Manuscript

Qian OUYANG, Yanfei HUANG, Lingyun YIN, Lingpeng ZHANG, Jing LIANG, Hongbin YU. Mechanism of HDAC3-Targeted Regulation of GATA-2 Affecting Osteogenic Differentiation of Dental Pulp Stem Cells[J]. Journal of Kunming Medical University.

Citation:

PDF( 3164 KB)

Mechanism of HDAC3-Targeted Regulation of GATA-2 Affecting Osteogenic Differentiation of Dental Pulp Stem Cells

Department of Stomatology，Yan’an Hospital Affiliated to Kunming Medical University，Kunming Yunnan 650500，China

Received Date: 2025-05-05

Abstract

Abstract

Objective To investigate the mechanism of the HDAC3/GATA-2 molecular axis in osteogenic differentiation of human dental pulp stem cells （hDPSCs）. Methods The hDPSCs cell line was cultured in osteogenic induction medium （OM）. Plasmids of oe-NC, oe-HDAC3 and oe-GATA-2 were constructed and transfected into the cells. Transfection efficiency was assessed by RT-qPCR and Western blot. Alkaline phosphatase （ALP） staining and alizarin red S （ARS） staining were used to evaluate ALP activity and mineralization nodules in cells. RT-qPCR and Western Blot were used to detect the expression levels of osteogenic key indicators COL1A1, ALP, BMP2, and RUNX2. GST-pull down and Co-Immunoprecipitation （CO-IP） were used to validate the interaction of HDAC3 and GATA-2 interaction, and immunofluorescence （IF） staining was performed to detect cellular expression. Results HDAC3 expression was significantly reduced during osteogenic differentiation of hDPSCs （P < 0.0001）. Overexpression of HDAC3 significantly attenuated ALP staining and activity （P = 0.0001）, as well as reduced the production of mineralized nodules in hDPSCs cells. The expression levels of COL1A1 （P = 0.0029）, ALP （P = 0.0001）, BMP2 （P = 0.0001） and RUNX2 （P = 0.0007） were inhibited after overexpression of HDAC3. HDAC3 interacted with GATA-2, and overexpression of HDAC3 decreased the expression of GATA-2 in cells （P = 0.0028）. Co-transfection of oe-GATA-2 reversed the inhibitory effects of HDAC3 overexpression, increasing ALP staining and activity （P = 0.0043） and mineralization nodules in cells. In the oe-HDAC3 + oe-GATA-2 group, the expression levels of COL1A1 （P = 0.0001）, ALP （P = 0.0423）, BMP2 （P < 0.0001）, and RUNX2 （P = 0.0005） were further elevated. Conclusion HDAC3 inhibited the osteogenic differentiation of hDPSCs by targeting and negatively regulating the expression of GATA-2.
- Human dental pulp stem cells,
- Osteogenic differentiation,
- HDAC3,
- GATA-2

FullText(HTML)

References(29)

References

[1]	Han Y, You X, Xing W, et al. Paracrine and endocrine actions of bone-the functions of secretory proteins from osteoblasts, osteocytes, and osteoclasts[J]. Bone Res, 2018, 6: 16. doi: 10.1038/s41413-018-0019-6
[2]	Fernandes G, Yang S. Application of platelet-rich plasma with stem cells in bone and periodontal tissue engineering[J]. Bone Res, 2016, 4: 16036. doi: 10.1038/boneres.2016.36
[3]	Ambrosi T H, Scialdone A, Graja A, et al. Adipocyte accumulation in the bone marrow during obesity and aging impairs stem cell-based hematopoietic and bone regeneration[J]. Cell Stem Cell, 2017, 20(6): 771-784. e6.
[4]	Chen X Y, Xu S Z, Wang X W, et al. Systematic comparison of biologically active foreign ions-codoped calcium phosphate microparticles on osteogenic differentiation in rat osteoporotic and normal mesenchymal stem cells[J]. Oncotarget, 2017, 8(22): 36578-36590. doi: 10.18632/oncotarget.16618
[5]	Camargo W A, de Vries R, van Luijk J, et al. Diabetes mellitus and bone regeneration: A systematic review and meta-analysis of animal studies[J]. Tissue Eng Part B Rev, 2017, 23(5): 471-479. doi: 10.1089/ten.teb.2016.0370
[6]	Yamada Y, Ito K, Nakamura S, et al. Promising cell-based therapy for bone regeneration using stem cells from deciduous teeth, dental pulp, and bone marrow[J]. Cell Transplant, 2011, 20(7): 1003-1013. doi: 10.3727/096368910X539128
[7]	Ledesma-Martínez E, Mendoza-Núñez V M, Santiago-Osorio E. Mesenchymal stem cells derived from dental pulp: A review[J]. Stem Cells Int, 2016, 2016: 4709572. doi: 10.1155/2016/4709572
[8]	Fu X, Jin L, Ma P, et al. Allogeneic stem cells from deciduous teeth in treatment for periodontitis in miniature swine[J]. J Periodontol, 2014, 85(6): 845-851. doi: 10.1902/jop.2013.130254
[9]	Ge X, Li Z, Jing S, et al. Parathyroid hormone enhances the osteo/odontogenic differentiation of dental pulp stem cells via ERK and P38 MAPK pathways[J]. J Cell Physiol, 2020, 235(2): 1209-1221. doi: 10.1002/jcp.29034
[10]	Edderkaoui M, Xu S, Chheda C, et al. HDAC3 mediates smoking-induced pancreatic cancer[J]. Oncotarget, 2016, 7(7): 7747-7760. doi: 10.18632/oncotarget.6820
[11]	Cantley M D, Bartold P M, Marino V, et al. Histone deacetylase inhibitors and periodontal bone loss[J]. J Periodontal Res, 2011, 46(6): 697-703. doi: 10.1111/j.1600-0765.2011.01392.x
[12]	Schroeder T M, Kahler R A, Li X, et al. Histone deacetylase 3 interacts with runx2 to repress the osteocalcin promoter and regulate osteoblast differentiation[J]. J Biol Chem, 2004, 279(40): 41998-42007. doi: 10.1074/jbc.M403702200
[13]	Ling K W, Ottersbach K, van Hamburg J P, et al. GATA-2 plays two functionally distinct roles during the ontogeny of hematopoietic stem cells[J]. J Exp Med, 2004, 200(7): 871-882. doi: 10.1084/jem.20031556
[14]	Rodrigues N P, Tipping A J, Wang Z, et al. GATA-2 mediated regulation of normal hematopoietic stem/progenitor cell function, myelodysplasia and myeloid leukemia[J]. Int J Biochem Cell Biol, 2012, 44(3): 457-460. doi: 10.1016/j.biocel.2011.12.004
[15]	Kamata M, Okitsu Y, Fujiwara T, et al. GATA2 regulates differentiation of bone marrow-derived mesenchymal stem cells[J]. Haematologica, 2014, 99(11): 1686-1696. doi: 10.3324/haematol.2014.105692
[16]	Lee E C, Kim Y M, Lim H M, et al. The histone deacetylase inhibitor (MS-275) promotes differentiation of human dental pulp stem cells into odontoblast-like cells independent of the MAPK signaling system[J]. Int J Mol Sci, 2020, 21(16): 57-71.
[17]	任小华. LncRNA LOC101928855调控炎性环境下牙髓干细胞成骨分化的机制研究[D]. 成都: 电子科技大学, 2022.
[18]	Cakouros D, Gronthos S. Epigenetic regulation of bone marrow stem cell aging: Revealing epigenetic signatures associated with hematopoietic and mesenchymal stem cell aging[J]. Aging Dis, 2019, 10(1): 174-189. doi: 10.14336/AD.2017.1213
[19]	Xu S, De Veirman K, Evans H, et al. Effect of the HDAC inhibitor vorinostat on the osteogenic differentiation of mesenchymal stem cells in vitro and bone formation in vivo[J]. Acta Pharmacol Sin, 2013, 34(5): 699-709. doi: 10.1038/aps.2012.182
[20]	Paino F, La Noce M, Tirino V, et al. Histone deacetylase inhibition with valproic acid downregulates osteocalcin gene expression in human dental pulp stem cells and osteoblasts: Evidence for HDAC2 involvement[J]. Stem Cells, 2014, 32(1): 279-289. doi: 10.1002/stem.1544
[21]	Li S, Li M, Liu X, et al. Genetic and chemical screenings identify HDAC3 as a key regulator in hepatic differentiation of human pluripotent stem cells[J]. Stem Cell Reports, 2018, 11(1): 22-31. doi: 10.1016/j.stemcr.2018.05.001
[22]	Bao Y, Chen H, Cai Z, et al. Advanced glycation end products inhibit neural stem cell differentiation via upregulation of HDAC3 expression[J]. Brain Res Bull, 2020, 159: 1-8. doi: 10.1016/j.brainresbull.2020.03.001
[23]	Jonason J H, Xiao G, Zhang M, et al. Post-translational regulation of Runx2 in bone and cartilage[J]. J Dent Res, 2009, 88(8): 693-703. doi: 10.1177/0022034509341629
[24]	Man K, Lawlor L, Jiang L H, et al. The selective histone deacetylase inhibitor MI192 enhances the osteogenic differentiation efficacy of human dental pulp stromal cells[J]. Int J Mol Sci, 2021, 22(10): 5224. doi: 10.3390/ijms22105224
[25]	Ozawa Y, Towatari M, Tsuzuki S, et al. Histone deacetylase 3 associates with and represses the transcription factor GATA-2[J]. Blood, 2001, 98(7): 2116-2123. doi: 10.1182/blood.V98.7.2116
[26]	Janardhan H P, Milstone Z J, Shin M, et al. Hdac3 regulates lymphovenous and lymphatic valve formation[J]. J Clin Invest, 2017, 127(11): 4193-4206. doi: 10.1172/JCI92852
[27]	Xu Y, Takahashi Y, Wang Y, et al. Downregulation of GATA-2 and overexpression of adipogenic gene-PPARgamma in mesenchymal stem cells from patients with aplastic Anemia[J]. Exp Hematol, 2009, 37(12): 1393-1399. doi: 10.1016/j.exphem.2009.09.005
[28]	Yamane T, Kunisada T, Yamazaki H, et al. Sequential requirements for SCL/tal-1, GATA-2, macrophage colony-stimulating factor, and osteoclast differentiation factor/osteoprotegerin ligand in osteoclast development[J]. Exp Hematol, 2000, 28(7): 833-840. doi: 10.1016/S0301-472X(00)00175-2
[29]	任明诗, 丁羽, 李子涵, 等. 成骨细胞与破骨细胞相互调节作用的研究进展[J]. 中国药理学通报, 2022, 38(6): 822-827.

Relative Articles(20)

Relative Articles

[1]	Hongyan CUI, Shuyang ZHANG, Wenwen ZHOU. Application of Serum STAT3，HDAC2，and Del-1 Levels in the Evaluation of Disease Severity and Prognosis of Children with Respiratory Syncytial Virus Pneumonia. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20251116
[2]	Yanfei HUANG, Hongbin YU, Lingyun YIN, Jing LIANG, Changquan LI, Dehong LI, Jinyuan WANG, Qian OUYANG. Regulatory Mechanism of Keap1/Nfe2L2 on Osteogenic Differentiation in Periodontitis. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20250704
[3]	Kun ZHOU, Yali LIU, Ziliang LI, Liping QIAN, Liquan RAN, Yalan REN. Effect of miR-34a on Proliferation and Osteogenic Differentiation of Human Periodontal Stem Cells. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20250403
[4]	Jingxiao XU, Jia LIU, Shu YAO, Xi ZHANG, Jiang LI, Guiqin CUI, Xiaoling YI, Dongyun LI. Effects and Mechanism of Quercetin on Osteogenic Differentiation of BMSCs. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20250504
[5]	Long PU, Xuanran ZHOU, Chenrong JIANG, Yunxuan LI, Yong YUAN. Effect of Inhibiting PPARγ Expression on Osteogenic Differentiation of BMSCs. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20240903
[6]	Shibing QIAN, Huiping SHI, Yanqiu LI, Rongyu YANG, Kaiwen DUAN. Research Progress on Osteogenic Differentiation of Apical Papilla Stem Cells. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20240926
[7]	Shibing QIAN, Lingpeng ZHANG, Lingyun YIN, Changquan LI, Hu LI, Hongbin YU. Study on Osteogenic Ability of Dental Pulp Stem Cells at Different Concentrations. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20230214
[8]	Qian XU, Yumei CUI, Siming MA, Yunhong LIN, Yijing XIONG, Zijun SONG, Xudong LI. miR-148a-3p Targeting SMURF2 in Regulating Osteogenic Differentiation and Enamel Development during In Vitro Tooth Organogenesis. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20231103
[9]	Yuxiao LI, Ming HE, Tianrong WANG, Yonggang YAO, Yu WANG. In Vitro Regulation of Th1/Th2 Cell of PBMCs in AR Mice by si-GATA-3. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20220802
[10]	Jing ZHOU, Ping XIONG, Chao-feng LIU, Li-qiong CHEN, Yong-hui ZHANG, Ke SHI, Yan NIE, Yan LIU. Periodontal Ligament Stem Cell BMP-2-PSH Composite Membrane in Repairing Alveolar Bone Defect in New Zealand Rabbits. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20210503
[11]	Liu Chong , Cao Hui , Yang Cai Cai , Wang Zhen . . Journal of Kunming Medical University,
[12]	Ji Qin Long , Kong Xiang Dong , Qi Shan Hong , Li Chao , Fan Jun , Sha Yong , Wang Shao Feng . The Experimental Study of Cyasterone on the Treatment of Osteoporosis through the Bidirectional Effect of Inhibiting Osteoclast Differetiation and Promoting Osteoblast Differentiation. Journal of Kunming Medical University,
[13]	Hu Zheng Xiong . Effect of TGF-β2 and geneX on the Proliferation and Osteogenic Differentiation of BrdU-labeled Bone Mesenchymal Stem Cells. Journal of Kunming Medical University,
[14]	Han Qun Chao . . Journal of Kunming Medical University,
[15]	Yao Han Xi . . Journal of Kunming Medical University,
[16]	Qi Zong Ze . . Journal of Kunming Medical University,
[17]	Wu Bin . . Journal of Kunming Medical University,
[18]	. Inducing Differentiation of Pig's Bone Mesenchymal Stem Cell(BMSC) to Urothelial Epithelia in Vitro. Journal of Kunming Medical University,
[19]	Mao Xi Hong . Pathological Study of PNS in Regulating Cathepsin B Expression in Rabbits with Pulmonary Fibrosis of Pulmonary Heart Disease. Journal of Kunming Medical University,
[20]	Li Wei Guo . . Journal of Kunming Medical University,