Background: We have previously demonstrated a Toll-like receptor (TLR)-mediated hyper-responsive phenotype in our cohort of localized aggressive periodontitis (LAP) individuals. However, mechanisms related to this phenotype are still not clear in the literature. The objective of this cross-sectional study is to examine the role of epigenetic regulation, specifically DNA methylation status of genes in the TLR pathway in this cohort. Peripheral blood was collected from 20 LAP patients and 20 healthy unrelated controls. Whole blood was stimulated with 1 μl (100 ng/μl) of purified Escherichia coli lipopolysaccharide (LPS) for 24 h and cyto/chemokines in the supernatants analyzed by Luminex multiplex assays. Genomic DNA extracted from buffy coats prepared from a second tube of whole blood was used for DNA methylation analysis by pyrosequencing of seven TLR signaling genes (FADD, MAP3K7, MYD88, IL6R, PPARA, IRAK1BP1, RIPK2). Results: Significant differences in the methylation status were observed at specific CpG positions in LAP patients compared to healthy controls and interestingly also between severe and moderate LAP. Specifically, subjects with moderate LAP presented hypermethylation of both the upregulating (MAP3K7, MYD88, IL6R, and RIPK2) and downregulating (FADD, IRAK, and PPARA) genes, while severe LAP presented hypomethylation of these genes. Further analysis on CpG sites with significant differences in methylation status correlates with an increased pro-inflammatory cytokine profile for LAP patients. Conclusions: Our findings suggest that epigenetic modifications of genes in the TLR pathway may orchestrate the thresholds for balancing induction and prevention of tissue destruction during the course of disease, and thus differ significantly at different stages of the disease, where moderate LAP shows hypermethylation and severe LAP shows hypomethylation of several genes. Trial registration:https://clinicaltrials.gov
|Published - Sep 2 2017
Bibliographical noteFunding Information:
This work was supported by the National Institutes of Health, NIH/NIDCR, under grant nos. R01DE019456 and R90DE22530-4.
The authors would like to thank the financial support for this research provided by NIH/NIDCR R01DE019456, as well as the doctors and staff at the Leon, Duval, and Jackson County Health Department dental centers for their assistance in coordinating our visits to their clinics, patient referrals, and dental care. This study is registered in Clinicaltrials.gov (#NCT01330719).
© 2017 The Author(s).
- Aggressive periodontitis
- Toll-like receptors
ASJC Scopus subject areas
- Molecular Biology
- Developmental Biology