The Histone Demethylase Inhibitor GSK-J4 Attenuates Periodontal Bone Loss and Inflammation in a Rat Model of Periodontitis

Abstract

Objective

To investigate the treatment effect of the histone demethylase inhibitor GSK-J4, a small molecule that inhibits the demethylase activity of Jumonji domain-containing protein 3 (JMJD3), in the treatment of periodontitis.

Methods

Gingival tissues from patients with moderate to severe chronic periodontitis and healthy controls were collected to evaluate JMJD3 expression via real-time quantitative reverse transcription PCR (RT-qPCR) and immunohistochemistry (IHC). Next, Sprague-Dawley (SD) rats were used to investigate the effect of GSK-J4 in vivo. The experimental periodontitis model was induced by upper first molar ligation and gingival sulcus injection of Porphyromonas gingivalis.The rats were divided into a healthy group, a periodontitis group, periodontitis plus GSK-J4 treatment groups (P + GSK-J4 15 mg/kg or 25 mg/kg), and a periodontitis plus dimethyl sulfoxide (DMSO) group (P + DMSO). After 4 weeks, maxillary molar segments were assessed via micro-computed tomography (CT) and hematoxylin and eosin (HE) staining. Serum tumor necrosis factor-α (TNF-α) levels were measured by enzyme-linked immunosorbent assay (ELISA).

Results

Higher expression of the Jmjd3gene and JMJD3 protein was detected in human inflamed gingiva than in healthy gingiva (P < 0.05). GSK-J4 administration reversed alveolar bone absorption [i.e., reduced alveolar bone crest (ABC)cementoenamel junction (CEJ) distance], reduced inflammatory cell accumulation at the crest of the alveolar bone, and alleviated serum TNF-α levels in rats with periodontitis. Moreover, the number of H3K27me3-positive nuclei was greater in model rats treated with GSK J4 than in model rats.

Conclusions

The histone demethylase inhibitor GSK-J4 attenuated periodontal bone loss and inflammation in a rat periodontitis model by targeting JMJD3.

Keywords

Periodontitis / Epigenetic modifications of histones / JMJD3 / Histone demethylase inhibitor / GSK-J4

© The Author(s), under exclusive licence to Huazhong University of Science and Technology 2025

Acknowledgements The authors thank Mr. Jian-wei Shang, Ms. Xinna Zhang and Ms. Wan-ying Qian for helping in the process of histological specimens’ preparation and staining.

Author Contributions Jian Kang: The first and corresponding author; drafting the manuscript, responsible for study design and carrying out the studies, analyzing data and completing interpretation; final approval of the version to be published. Huan Yu: carrying out the experiments, collecting the data, drafting and revising the manuscript for improvement of English language. Xu Xiang: participating in study design, data analysis and interpretation; drafting and revising the manuscript, final approval of the version to be published. Yong-qiang Ma, Le Zhang and Yuan Zhang: carrying out the experiments, performing the statistical analysis. Zhi-tao Wang and Jing Yang Zheng Zhang: participating in acquisition, analysis, or interpretation of data. Hui-ru Zou and Yue Wang: substantial contributions to study design, preparing the graphs in this article, revising the manuscript. All authors read and approved the final manuscript.

Funding This study was supported by the Tianjin Stomatological Hospital MD and PhD Key Program (No. 2019BSZD11), Periodontal Key Discipline Project of Tianjin Stomatological Hospital (2022P02), the Science and Technology Project of Tianjin Health Commission (No. ZC20039), and the High-level Talents in the Medical/Health Care Industry—Young Medical Elites (No. TJSQNYXXR-D2-114).

Data Availability All data generated or analysed during this study are included in this published article.

Declarations

Conflict of Interest The authors have no conflicts of interest to declare.

Ethical Approval and Consent to Participate This study using rats as research subjects is approved by Tianjin Stomatological Hospital, approval number (PA2019-B-006).

Consent for Publication All the authors approved the submission and publication of this manuscript.

Human Ethics This study protocol using human gingiva was reviewed and approved by Tianjin Stomatological Hospital, approval number PH2019-B-005. Informed consents were obtained from all the patients.

References

[1] Kassebaum NJ, Bernabe E, Dahiya M, et al. Global burden of severe periodontitis in 1990-2010: a systematic review and metaregression. J Dent Res. 2014; 93(11):1045-1053.

[2] Eke PI, Dye BA, Wei L, et al. Prevalence of periodontitis in adults in the United States: 2009 and 2010. J Dent Res. 2012; 91(10): 914-920.

[3] Mosaddad SA, Hussain A, Tebyaniyan H. Green alternatives as antimicrobial agents in mitigating periodontal diseases: a narrative review. Microorganisms. 2023; 11(5):1269.

[4] Kwon T, Lamster IB, Levin L. Current concepts in the management of periodontitis. Int Dent J. 2021; 71(6):462-476.

[5] Khayatan D, Bagherzadeh Oskouei A, Alam M, et al. Cross talk between cells and the current bioceramics in bone regeneration: a comprehensive review. Cell Transplant. 2024; 33:9636897241236030.

[6] Jurdzinski KT, Potempa J, Grabiec AM. Epigenetic regulation of inflammation in periodontitis: cellular mechanisms and therapeutic potential. Clin Epigenetics. 2020; 12(1):186.

[7] Hussain A, Tebyaniyan H, Khayatan D. The role of epigenetic in dental and oral regenerative medicine by different types of dental stem cells: a comprehensive overview. Stem Cells Int. 2022; 2022:5304860.

[8] Larsson L, Castilho RM, Giannobile WV. Epigenetics and its role in periodontal diseases: a state-of-the-art review. J Periodontol. 2015; 86(4):556-568.

[9] Liaw A, Liu C, Ivanovski S, et al. The relevance of DNA methylation and histone modification in periodontitis: a scoping review. Cells. 2022; 11(20):3211.

[10] Barros SP, Offenbacher S. Modifiable risk factors in periodontal disease: epigenetic regulation of gene expression in the inflammatory response. Periodontology 2000. 2014; 64(1):95-110.

[11] Martins MD, Jiao Y, Larsson L, et al. Epigenetic modifications of histones in periodontal disease. J Dent Res. 2016; 95(2):215-222.

[12] Zhang X, Liu L, Yuan X, et al. JMJD3 in the regulation of human diseases. Protein Cell. 2019; 10(12):864-882.

[13] Yasui T, Hirose J, Tsutsumi S, et al. Epigenetic regulation of osteoclast differentiation: possible involvement of Jmjd3 in the histone demethylation of Nfatc1. J Bone Miner Res. 2011; 26(11):2665-2671.

[14] Shan J, Fu L, Balasubramanian MN, et al. ATF4-dependent regulation of the JMJD3 gene during amino acid deprivation can be rescued in Atf4-deficient cells by inhibition of deacetylation. J Biol Chem. 2012; 287(43):36393-36403.

[15] Lee HY, Choi K, Oh H, et al. HIF-1-dependent induction of Jumonji domain-containing protein (JMJD) 3 under hypoxic conditions. Mol Cells. 2014, 37(1):43-50.

[16] Hui T, A P, Zhao Y, et al. EZH2, a potential regulator of dental pulp inflammation and regeneration. J Endod. 2014; 40(8):1132-1138.

[17] Wang P, Yue J, Xu W, et al. Jumonji domain-containing protein 3 regulates the early inflammatory response epigenetically in human periodontal ligament cells. Arch Oral Biol. 2018; 93:87-94.

[18] Jin Y, Liu Z, Li Z, et al. Histone demethylase JMJD3 downregulation protects against aberrant force-induced osteoarthritis through epigenetic control of NR4A1. Int J Oral Sci. 2022; 14(1):34.

[19] Chen YH, Chen CH, Chien CY, et al. JMJD3 suppresses tumor progression in oral tongue squamous cell carcinoma patients receiving surgical resection. PeerJ. 2022; 10:e13759.

[20] An C, Jiao B, Du H, et al. Jumonji domain-containing protein-3 (JMJD3) promotes myeloid fibroblast activation and macrophage polarization in kidney fibrosis. Br J Pharmacol. 2023; 180(17):2250-2265.

[21] Gao Y, Yu W, Song J, et al. JMJD3 ablation in myeloid cells confers renoprotection in mice with DOCA/salt-induced hypertension. Hypertens Res. 2023; 46(8):1934-1948.

[22] Donas C, Carrasco M, Fritz M, et al. The histone demethylase inhibitor GSK-J4 limits inflammation through the induction of a tolerogenic phenotype on DCs. J Autoimmun. 2016; 75:105-117.

[23] Chen D, Cai B, Zhu Y, et al. Targeting histone demethylases JMJD3 and UTX: selenium as a potential therapeutic agent for cervical cancer. Clin Epigenetics. 2024; 16(1):51.

[24] Dalpatraj N, Naik A, Thakur N. GSK-J4: an H3K27 histone demethylase inhibitor, as a potential anti-cancer agent. Int J Cancer. 2023; 153(6):1130-1138.

[25] Kruidenier L, Chung CW, Cheng Z, et al. A selective jumonji H3K27 demethylase inhibitor modulates the proinflammatory macrophage response. Nature. 2012; 488(7411):404-408.

[26] Jia W, Wu W, Yang D, et al. Histone demethylase JMJD3 regulates fibroblast-like synoviocyte-mediated proliferation and joint destruction in rheumatoid arthritis. FASEB J. 2018; 32(7):4031-4042.

[27] Jun Z, Xinmeng J, Yue L, et al. Jumonji domain containing-3 (JMJD3) inhibition attenuates IL-1β-induced chondrocytes damage in vitro and protects osteoarthritis cartilage in vivo. Inflamm Res. 2020; 69(7):657-666.

[28] Kirkpatrick JE, Kirkwood KL, Woster PM. Inhibition of the histone demethylase KDM4B leads to activation of KDM1A, attenuates bacterial-induced pro-inflammatory cytokine release, and reduces osteoclastogenesis. Epigenetics. 2018; 13(5):557-572.

[29] Yang H, Wen Q, Xue J, et al. Alveolar bone regeneration potential of a traditional Chinese medicine, Bu-Shen-GuChi-Wan, in experimental periodontitis. J Periodontal Res. 2014; 49(3):382-389.

[30] Cantley MD, Bartold PM, Marino V, et al. Histone deacetylase inhibitors and periodontal bone loss. J Periodontal Res. 2011; 46(6):697-703.

[31] Chen Y, Liu Z, Pan T, et al. JMJD 3 is involved in neutrophil membrane proteinase 3 overexpression during the hyperinflammatory response in early sepsis. Int Immunopharmacol. 2018; 59:40-46.

[32] Chen Y, Liu W, Xu X, et al. The role of H3K27me3-mediated Th 17 differentiation in ankylosing spondylitis. 2024; 47(5):1685-1698.

[33] Park CS, Lee JY, Choi HY, et al. Gallic acid attenuates bloodspinal cord barrier disruption by inhibiting Jmjd3 expression and activation after spinal cord injury. Neurobiol Dis. 2020; 145:105077.

[34] Davis FM, Tsoi LC, Melvin WJ, et al. Inhibition of macrophage histone demethylase JMJD3 protects against abdominal aortic aneurysms. J Exp Med. 2021; 218(6):e20201839.

[35] Yapp C, Carr AJ, Price A, et al. H3K27me3 demethylases regulate in vitro chondrogenesis and chondrocyte activity in osteoarthritis. Arthritis Res Ther. 2016; 18(1):158.

[36] Huang D, Zhang C, Wang P, et al. JMJD 3 promotes Porphyromonas gingivalis lipopolysaccharide-induced Th17-cell differentiation by modulating the STAT3-RORc signaling pathway. DNA Cell Biol. 2022; 41(8):778-787.

[37] de Camargo Pereira G, Guimaraes GN, Planello AC, et al. Porphyromonas gingivalis LPS stimulation downregulates DNMT1, DNMT3a, and JMJD 3 gene expression levels in human HaCaT keratinocytes. Clin Oral Investig. 2013; 17(4):1279-1285.

[38] Yin L, Chung WO. Epigenetic regulation of human beta-defensin 2 and CC chemokine ligand 20 expression in gingival epithelial cells in response to oral bacteria. Mucosal Immunol. 2011; 4(4):409-419.

[39] Jimi E, Fei H, Nakatomi C. NF-κB signaling regulates physiological and pathological chondrogenesis. Int J Mol Sci. 2019; 20(24):6275.

[40] Madureira DF, et al.Lucas De Abreu Lima I, Costa GC, Tumor necrosis factor-alpha in gingival crevicular fluid as a diagnostic marker for periodontal diseases: a systematic review. J Evid Based Dent Pract. 2018; 18(4):315-331.

[41] Górska R, Gregorek H, Kowalski J, et al. Relationship between clinical parameters and cytokine profiles in inflamed gingival tissue and serum samples from patients with chronic periodontitis. J Clin Periodontol. 2003; 30(12):1046-1052.

[42] Kawai T, Matsuyama T, Hosokawa Y, et al. B and T lymphocytes are the primary sources of RANKL in the bone resorptive lesion of periodontal disease. Am J Pathol. 2006; 169(3):987-998.

[43] Preshaw PM. Host modulation therapy with anti-inflammatory agents. Periodontology 2000. 2018; 76(1):131-149.

[44] Xuan D, Han Q, Tu Q, et al. Epigenetic modulation in periodontitis: interaction of adiponectin and JMJD3-IRF4 axis in macrophages. J Cell Physiol. 2016; 231(5):1090-1096.