Damage-regulated autophagy modulator 1 in oral inflammation and infection
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Damage-regulated autophagy modulator (DRAM) 1 is a p53 target gene with possible involvement in oral inflammation and infection. This study sought to examine the presence and regulation of DRAM1 in periodontal diseases.
Material and methods
In vitro, human periodontal ligament fibroblasts were exposed to interleukin (IL)-1β and Fusobacterium nucleatum for up to 2 days. The DRAM1 synthesis and its regulation were analyzed by real-time PCR, immunocytochemistry, and ELISA. Expressions of other autophagy-associated genes were also studied by real-time PCR. In vivo, synthesis of DRAM1 in gingival biopsies from rats and patients with and without periodontal disease was examined by real-time PCR and immunohistochemistry. For statistics, ANOVA and post-hoc tests were applied (p < 0.05).
In vitro, DRAM1 was significantly upregulated by IL-1β and F. nucleatum over 2 days and a wide range of concentrations. Additionally, increased DRAM1 protein levels in response to both stimulants were observed. Autophagy-associated genes ATG3, BAK1, HDAC6, and IRGM were also upregulated under inflammatory or infectious conditions. In vivo, the DRAM1 gene expression was significantly enhanced in rat gingival biopsies with induced periodontitis as compared to control. Significantly increased DRAM1 levels were also detected in human gingival biopsies from sites of periodontitis as compared to healthy sites.
Our data provide novel evidence that DRAM1 is increased under inflammatory and infectious conditions in periodontal cells and tissues, suggesting a pivotal role of DRAM1 in oral inflammation and infection.
DRAM1 might be a promising target in future diagnostic and treatment strategies for periodontitis.
KeywordsDamage-regulated autophagy modulator Autophagy Fusobacterium nucleatum Interleukin-1β Periodontal ligament Periodontitis
The authors would like to thank Ms. Ramona Menden, Ms. Silke van Dyck, Ms. Inka Bay and Prof. Heiko Spallek for their valuable support.
This study was supported by the Medical Faculty of the University of Bonn, the University of Sydney, the German Orthodontic Society (DGKFO), the German Research Foundation (DFG, ME 4798/1–1) and by the German Federal Ministry of Education and Research (grant 01EK1603A-Neuro2D3, to AT).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Approval of the Ethics Committee of the University of Bonn was obtained (#117/15 #043/11) and of the Ethical Committee on Animal Experimentation (protocol number: 23/2012) from the School of Dentistry at Araraquara. Animal experiments were carried out following the recommendations of the ARRIVE guidelines.
All donors of the PDL cells or their parents gave written informed consent.
- 10.Bullon P, Cordero MD, Quiles JL, Ramirez-Tortosa Mdel C, Gonzalez-Alonso A, Alfonsi S, García-Marín R, de Miguel M, Battino M (2012) Autophagy in periodontitis patients and gingival fibroblasts: unraveling the link between chronic diseases and inflammation. BMC Med 10:122. https://doi.org/10.1186/1741-7015-10-122 CrossRefPubMedPubMedCentralGoogle Scholar
- 24.Nokhbehsaim M, Keser S, Nogueira AV, Cirelli JA, Jepsen S, Jäger A, Eick S, Deschner J (2014) Beneficial effects of adiponectin on periodontal ligament cells under normal and regenerative conditions. J Diabetes Res 2014:796565. https://doi.org/10.1155/2014/796565 CrossRefPubMedPubMedCentralGoogle Scholar
- 26.Memmert S, Damanaki A, Nogueira AVB, Eick S, Nokhbehsaim M, Papadopoulou AK, Till A, Rath B, Jepsen S, Götz W, Piperi C, Basdra EK, Cirelli JA, Jäger A, Deschner J (2017) Role of Cathepsin S in periodontal inflammation and infection. Mediat Inflamm 2017:4786170. https://doi.org/10.1155/2017/4786170 CrossRefGoogle Scholar
- 29.Memmert S, Gölz L, Pütz P, Jäger A, Deschner J, Appel T, Baumgarten G, Rath-Deschner B, Frede S, Götz W (2015) Regulation of p53 under hypoxic and inflammatory conditions in periodontium. Clin Oral Investig 20:1781–1789. https://doi.org/10.1007/s00784-015-1679-x CrossRefPubMedPubMedCentralGoogle Scholar
- 34.Saito A, Inagaki S, Kimizuka R, Okuda K, Hosaka Y, Nakagawa T, Ishihara K (2008) Fusobacterium nucleatum enhances invasion of human gingival epithelial and aortic endothelial cells by Porphyromonas gingivalis. FEMS Immunol Med Microbiol 54:349–355. https://doi.org/10.1111/j.1574-695X.2008.00481.x CrossRefPubMedPubMedCentralGoogle Scholar
- 35.He J, Huang W, Pan Z, Cui H, Qi G, Zhou X, Chen H (2012) Quantitative analysis of microbiota in saliva, supragingival, and subgingival plaque of Chinese adults with chronic periodontitis. Clin Oral Investig 16:1579–1588. https://doi.org/10.1007/s00784-011-0654-4 CrossRefPubMedPubMedCentralGoogle Scholar
- 36.Signat B, Roques C, Poulet P, Duffaut D (2011) Fusobacterium nucleatum in periodontal health and disease. Curr Issues Mol Biol 13:25–36Google Scholar
- 46.Van der Vaart M, Korbee CJ, Lamers GE, Tengeler AC, Hosseini R, Haks MC, Ottenhoff TH, Spaink HP, Meijer AH (2014) The DNA damage-regulated autophagy modulator DRAM1 links mycobacterial recognition via TLR-MYD88 to autophagic defense [corrected]. Cell Host Microbe 15:753–767. https://doi.org/10.1016/j.chom.2014 CrossRefPubMedPubMedCentralGoogle Scholar