Sirt3 Rescues Porphyromonas gingivalis-Impaired Cementogenesis via SOD2 Deacetylation

Abstract

The keystone pathogen Porphyromonas gingivalis (P.g.) is responsible for cementum resorption in periodontitis; however, the mechanism involved in it remains unclear. Sirtuin 3 (Sirt3) is a NAD⁺-dependent protein deacetylase contributing to mitochondrial homeostasis and various cell functions. In this study, the expression of Sirt3 in cementoblasts was found to be increased during cementoblast mineralisation and cementum development, while it decreased gradually under P.g. infection in a multiplicity of infection-dependent manner. Compared with wild type mice, the Sirt3 knockout mice showed less cellular cementum and lower mineralisation capacity with decreased expression of Runx2 and OCN in cementoblasts. Sirt3 inhibition by 3-TYP or Sirt3 silencing by lentivirus infection both confirmed the impaired cementogenesis. Conversely, honokiol (HKL) was simulated to bind Sirt3 and was applied to activate Sirt3 in cementoblasts. HKL-mediated Sirt3 activation facilitated cementoblast mineralisation and rescued P.g.-suppressed cementoblast mineralisation markedly. Superoxide dismutase 2 (SOD2), the downstream molecule of Sirt3, showed a similar expression pattern to Sirt3 under different conditions. Silencing of SOD2 was demonstrated to restrain cementoblast mineralisation. The pan acetylation was detected to decrease under Sirt3-upregulating conditions and increase under Sirt3-downregulating conditions. The binding of Sirt3 and SOD2 in cementoblasts was also verified. Furthermore, SOD2 acetylation and specific SOD2-K68 acetylation were found to be upregulated under P.g. or Sirt3 silencing conditions and downregulated by HKL stimulation. Moreover, K68Q mutation simulating acetylation decreased cementoblast mineralisation, while K68R mutation simulating deacetylation increased it. Altogether, Sirt3 deacetylates SOD2 via K68 to orchestrate P.g.-perturbed cementogenesis, and HKL is a Sirt3-targeted treatment candidate.

Keywords

acetylation / cementogenesis / cementum / porphyromonas gingivalis / sirtuin3 / superoxide dismutase

References

[1] D. D. Bosshardt and K. A. Selvig, “Dental Cementum: The Dynamic Tissue Covering of the Root,” Periodontology 2000 13 (1997): 41-75.

[2] H. Arzate, M. Zeichner-David, and G. Mercado-Celis, “Cementum Proteins: Role in Cementogenesis, Biomineralization, Periodontium Formation and Regeneration,” Periodontology 2000 67, no. 1 (2015): 211-233.

[3] G.-p. Dang, Y. Wei, Q.-q. Wan, et al., “Regulatory Mechanisms and Regeneration Strategies of the Soft-Hard Tissue Interface in the Human Periodontium,” BMEMat 2, no. 3 (2024): e12069.

[4] J. Nuñez, F. Vignoletti, R. G. Caffesse, and M. Sanz, “Cellular Therapy in Periodontal Regeneration,” Periodontology 2000 79, no. 1 (2019): 107-116.

[5] G. Hajishengallis, R. P. Darveau, and M. A. Curtis, “The Keystone-Pathogen Hypothesis,” Nature Reviews. Microbiology 10, no. 10 (2012): 717-725.

[6] W. Wang, Z. Zhou, T. Ding, et al., “Capsaicin Attenuates Porphyromonas gingivalis-Suppressed Osteogenesis of Periodontal Ligament Stem Cells via Regulating Mitochondrial Function and Activating PI3K/AKT/mTOR Pathway,” Journal of Periodontal Research 59, no. 4 (2024): 798-811.

[7] X. Huang, Y. Deng, J. Xiao, H. Wang, Q. Yang, and Z. Cao, “Genetically Engineered M2-Like Macrophage-Derived Exosomes for P. gingivalis-Suppressed Cementum Regeneration: From Mechanism to Therapy,” Bioactive Materials 32 (2024): 473-487.

[8] X. Huang, L. Ma, X. Wang, et al., “Ckip-1 Mediates P. gingivalis-Suppressed Cementoblast Mineralization,” Journal of Dental Research 101, no. 5 (2022): 599-608.

[9] J. Zhang, H. Xiang, J. Liu, Y. Chen, R.-R. He, and B. Liu, “Mitochondrial Sirtuin 3: New Emerging Biological Function and Therapeutic Target,” Theranostics 10, no. 18 (2020): 8315-8342.

[10] M. Cao, Q. Zhao, X. Sun, et al., “Sirtuin 3: Emerging Therapeutic Target for Cardiovascular Diseases,” Free Radical Biology and Medicine 180 (2022): 63-74.

[11] Z. D. Zhou and E. K. Tan, “Oxidized Nicotinamide Adenine Dinucleotide-Dependent Mitochondrial Deacetylase Sirtuin-3 as a Potential Therapeutic Target of Parkinson's Disease,” Ageing Research Reviews 62 (2020): 101107.

[12] Y. Wang, M. Barthez, and D. Chen, “Mitochondrial Regulation in Stem Cells,” Trends in Cell Biology 34 (2023): 685-694.

[13] J. Gao, Z. Feng, X. Wang, et al., “SIRT3/SOD2 Maintains Osteoblast Differentiation and Bone Formation by Regulating Mitochondrial Stress,” Cell Death and Differentiation 25, no. 2 (2018): 229-240.

[14] C. Lambona, C. Zwergel, S. Valente, and A. Mai, “SIRT3 Activation a Promise in Drug Development? New Insights Into SIRT3 Biology and Its Implications on the Drug Discovery Process,” Journal of Medicinal Chemistry 67, no. 3 (2024): 1662-1689.

[15] K. Xu, J. Li, R. Wen, B. Chang, Y. Cheng, and X. Yi, “Role of SIRT3 in Bone Homeostasis and Its Application in Preventing and Treating Bone Diseases,” Frontiers in Pharmacology 14 (2023): 1248507.

[16] G. Li, H. Qin, M. Zhou, et al., “Knockdown of SIRT3 Perturbs Protective Effects of Irisin Against Bone Loss in Diabetes and Periodontitis,” Free Radical Biology and Medicine 200 (2023): 11-25.

[17] J. Chen, Y. Zhang, J. Gao, T. Li, X. Gan, and H. Yu, “Sirtuin 3 Deficiency Exacerbates Age-Related Periodontal Disease,” Journal of Periodontal Research 56, no. 6 (2021): 1163-1173.

[18] C. R. Salmon, D. M. Tomazela, K. G. S. Ruiz, et al., “Proteomic Analysis of Human Dental Cementum and Alveolar Bone,” Journal of Proteomics 91 (2013): 544-555.

[19] I. N. Zelko, T. J. Mariani, and R. J. Folz, “Superoxide Dismutase Multigene Family: A Comparison of the CuZn-SOD (SOD1), Mn-SOD (SOD2), and EC-SOD (SOD3) Gene Structures, Evolution, and Expression,” Free Radical Biology and Medicine 33, no. 3 (2002): 337-349.

[20] T. Fukai and M. Ushio-Fukai, “Superoxide Dismutases: Role in Redox Signaling, Vascular Function, and Diseases,” Antioxidants and Redox Signaling 15, no. 6 (2011): 1583-1606.

[21] K. Tsesmelis, G. Maity-Kumar, D. Croner, et al., “Accelerated Aging in Mice With Astrocytic Redox Imbalance as a Consequence of SOD2 Deletion,” Aging Cell 22, no. 9 (2023): e13911.

[22] F.-Y. Deng, S.-F. Lei, X.-D. Chen, L.-J. Tan, X.-Z. Zhu, and H.-W. Deng, “An Integrative Study Ascertained SOD2 as a Susceptibility Gene for Osteoporosis in Chinese,” Journal of Bone and Mineral Research 26, no. 11 (2011): 2695-2701.

[23] H. Kim, Y. D. Lee, H. J. Kim, Z. H. Lee, and H.-H. Kim, “SOD2 and Sirt3 Control Osteoclastogenesis by Regulating Mitochondrial ROS,” Journal of Bone and Mineral Research 32, no. 2 (2017): 397-406.

[24] E. A. Abou Neel, A. Aljabo, A. Strange, et al., “Demineralization-Remineralization Dynamics in Teeth and Bone,” International Journal of Nanomedicine 11 (2016): 4743-4763, https://doi.org/10.2147/IJN.S107624.

[25] J. Song, L. Li, L. Fang, et al., “Advanced Strategies of Scaffolds Design for Bone Regeneration,” BMEMat 1, no. 4 (2023): e12046.

[26] N. Zhao, B. L. Foster, and L. F. Bonewald, “The Cementocyte-An Osteocyte Relative?,” Journal of Dental Research 95, no. 7 (2016): 734-741.

[27] Y. Ding, H. Yang, Y. Wang, J. Chen, Z. Ji, and H. Sun, “Sirtuin 3 Is Required for Osteogenic Differentiation Through Maintenance of PGC-1ɑ-SOD2-Mediated Regulation of Mitochondrial Function,” International Journal of Biological Sciences 13, no. 2 (2017): 254-264.

[28] F. Liu, L. Yuan, L. Li, et al., “S-Sulfhydration of SIRT3 Combats BMSC Senescence and Ameliorates Osteoporosis via Stabilizing Heterochromatic and Mitochondrial Homeostasis,” Pharmacological Research 192 (2023): 106788.

[29] J.-E. Huh, J. H. Shin, E. S. Jang, et al., “Sirtuin 3 (SIRT3) Maintains Bone Homeostasis by Regulating AMPK-PGC-1β Axis in Mice,” Scientific Reports 6 (2016): 22511.

[30] J. Mysak, S. Podzimek, P. Sommerova, et al., “Porphyromonas gingivalis: Major Periodontopathic Pathogen Overview,” Journal of Immunology Research 2014 (2014): 476068.

[31] Z. Deng, M. He, H. Hu, et al., “Melatonin Attenuates Sepsis-Induced Acute Kidney Injury by Promoting Mitophagy Through SIRT3-Mediated TFAM Deacetylation,” Autophagy 20, no. 1 (2024): 151-165.

[32] Y. Zhang, Y. Liu, M. Hou, et al., “Reprogramming of Mitochondrial Respiratory Chain Complex by Targeting SIRT3-COX4I2 Axis Attenuates Osteoarthritis Progression,” Advanced Science 10, no. 10 (2023): e2206144.

[33] Z. Wang, R. Sun, G. Wang, et al., “SIRT3-Mediated Deacetylation of PRDX3 Alleviates Mitochondrial Oxidative Damage and Apoptosis Induced by Intestinal Ischemia/Reperfusion Injury,” Redox Biology 28 (2020): 101343.

[34] X. Liu, D. Li, W. Pi, et al., “LCZ696 Protects Against Doxorubicin-Induced Cardiotoxicity by Inhibiting Ferroptosis via AKT/SIRT3/SOD2 Signaling Pathway Activation,” International Immunopharmacology 113 (2022): 109379.

[35] J. He, X. Liu, C. Su, et al., “Inhibition of Mitochondrial Oxidative Damage Improves Reendothelialization Capacity of Endothelial Progenitor Cells via SIRT3 (Sirtuin 3)-Enhanced SOD2 (Superoxide Dismutase 2) Deacetylation in Hypertension,” Arteriosclerosis, Thrombosis, and Vascular Biology 39, no. 8 (2019): 1682-1698.

[36] L. Ning, X. Rui, L. Guorui, et al., “A Novel Mechanism for the Protection Against Acute Lung Injury by Melatonin: Mitochondrial Quality Control of Lung Epithelial Cells Is Preserved Through SIRT3-Dependent Deacetylation of SOD2,” Cellular and Molecular Life Sciences 79, no. 12 (2022): 610.

[37] J. Wang, J. Li, Y. He, et al., “The SIRT3 Activator Ganoderic Acid D Regulates Airway Mucin MUC5AC Expression via the NRF2/GPX4 Pathway,” Pulmonary Pharmacology and Therapeutics 83 (2023): 102262.

[38] X. Wang, T. Shen, J. Lian, et al., “Resveratrol Reduces ROS-Induced Ferroptosis by Activating SIRT3 and Compensating the GSH/GPX4 Pathway,” Molecular Medicine 29, no. 1 (2023): 137.

[39] A. Sharma, S. Sinha, H. Keswani, and N. Shrivastava, “Kaempferol and Apigenin Suppresses the Stemness Properties of TNBC Cells by Modulating Sirtuins,” Molecular Diversity 26, no. 6 (2022): 3225-3240.

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2025 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.