Epigenetics, DNA Methylation Patterns and Periodontal Disease (SZU)
The overall goal of this research is (1) to identify changes in gene expression and DNA methylation status in subjects who exhibit advanced chronic periodontal inflammation and (2) to identify microRNAs (miRNAs) and the interactive pathways associated with obesity as a modifier of periodontal infection pathogenesis.
|Study Design:||Observational Model: Case Control
Time Perspective: Cross-Sectional
|Official Title:||Host DNA Methylation Patterns in Association With Periodontal Disease : MiRNA Changes in Obesity|
- DNA methylation patterns of selected candidate genes [ Time Frame: Visit 1 ] [ Designated as safety issue: No ]
- Identification of miRNA species [ Time Frame: Visit 1 ] [ Designated as safety issue: No ]
Biospecimen Retention: Samples With DNA
- Gingiva biopsy samples (epithelium, connective tissue)
- Gingival crevicular fluid
- Plaque samples
|Study Start Date:||October 2007|
|Study Completion Date:||February 2011|
|Primary Completion Date:||February 2011 (Final data collection date for primary outcome measure)|
Obese group (+ periodontitis group) Non-obese group (+ periodontitis group)
DNA methylation study
Periodontitis group Healthy periodontium group
Aim 1. To determine whether diseased periodontal tissues biopsied from patients with periodontal disease will have altered DNA methylation patterns of selected genes as compared to the DNA methylation status biopsies obtained from adjacent, non-diseased periodontal sites, as well as periodontal biopsy samples obtained from non-diseased, healthy subjects.
Aim 2. To determine the role of tissue DNA Methylation on mRNA expression:
- 2a. To determine whether alterations in tissue mRNA expression (as determined by quantitative PCR) are associated with an aberrant DNA methylation status of respective gene promoter CpG islands in tissue biopsy samples obtained from diseased and non-diseased tissues from patients with periodontal disease and from non-diseased healthy subjects.
- 2b. By performing laser capture microdissection of biopsy samples from diseased patients [(epithelium, connective tissue and inflammatory infiltrate)] we seek to determine cellular patterns of mRNA expression and DNA methylation of targeted genes comparing inflamed to non-inflamed sites within diseased patients.
- 2c. We will perform in vitro studies to analyze whether gingival fibroblasts from patients with periodontal disease will show a differential methylation pattern and whether these cells will display a differential inflammatory response when compared to control fibroblasts isolated from healthy subjects.
Aim 3. The aim of this pilot investigation was to determine if miRNA expression differed in the presence or absence of obesity, comparing gingival biopsies obtained from patients with or without periodontal disease.
- 3a. To determine whether diseased periodontal tissues biopsied from patients with periodontal disease will influence the level of miRNA expression as compared to the biopsies obtained from non-diseased, healthy subjects.
- 3b. To determine whether BMI [nonobese subjects BMI <30kg/m2, obese subjects >30kg/m2] will influence the level of miRNA expression in biopsies from diseased periodontal tissues as compared to periodontal biopsy samples obtained from non-diseased, healthy subjects.
Gingival samples, gingival crevicular fluid, saliva and plaque samples will be collected and a periodontal examination will be performed. The periodontal assessment will record pocket depth, clinical attachment level and percent bleeding on probing. Blood pressure, height and weight (Body Mass Index assessment) will be obtained on participants as well.
Tissue gene expression by quantitative PCR and DNA methylation sequence analysis, proteomic analyses of gingival crevicular fluid, Differential Methylation Hybridization and, microRNA expression profile by Human miRNA Microarray and Real-time PCR quantification will all be performed.
|United States, North Carolina|
|Center for Oral and Systemic Diseases|
|Chapel Hill, North Carolina, United States, 27599|
|Principal Investigator:||Steven Offenbacher, DDS, MS, PhD||University of North Carolina, Chapel Hill|