Elsevier

Archives of Oral Biology

Volume 65, May 2016, Pages 44-51
Archives of Oral Biology

Determination of NLRP3 (rs4612666) and IL-1B (rs1143634) genetic polymorphisms in periodontally diseased and healthy subjects

https://doi.org/10.1016/j.archoralbio.2016.01.013Get rights and content

Highlights

  • Genetic factors might play a role in the etiopathogenesis of CP.

  • NLRP3 (rs4612666) and IL-1B (rs1143634) SNPs were genotyped in a Colombian population.

  • Nor NLRP3 (rs4612666) neither IL-1B (rs1143634) SNPs were independently associated with CP.

  • Age stratum/smoking habit showed synergistic interactions with CT/CC genotypes of NLRP3 gene.

  • Synergistic interaction might play a significant role in the pathogenic pathways of CP.

Abstract

Objective

The focus of the current study was to identify if a possible association between NLRP3 (rs4612666) and IL-1B (rs1143634) single-nucleotide polymorphisms (SNPs) may be implicated in the etiopathogenesis of chronic periodontitis (CP) in a Colombian population.

Design

One hundred and twenty-four CP subjects and 81 periodontally healthy controls (HC) were recruited. Periodontal status was assessed by criteria based on probing depth, clinical attachment level, extent, and severity of periodontal breakdown. Human genomic DNA was obtained from saliva samples of the study subjects. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to identify the NLRP3 (rs4612666) and IL-1B (rs1143634) SNPs. The association of polymorphisms with CP was assessed individually and adjusted for confounding using a multivariate binary logistic regression model.

Results

Bivariate analysis showed a weak association between CT genotype of NLRP3 (rs4612666) SNP and CP, however after logistic regression analysis, neither NLRP3 (rs4612666) nor IL-1B (rs1143634) polymorphisms were strongly/independently associated with disease status. Even so, an interaction effect was significantly detected not only among CT/CC genotypes of NLRP3 gene regarding to the age stratum ≥48 years, but also between CC genotype of the same gene and smoking habit.

Conclusion

Although the present results do not support that IL-1B (rs1143634) SNP could be identified as a risk predictor for CP in the present population, the synergistic interaction of the CT/CC genotypes of NLRP3 (rs4612666) SNP with ageing and/or smoking habit potentially might play a significant role in the pathogenic pathways of periodontal disease.

Introduction

Chronic periodontitis (CP), an infection-induced inflammatory and progressive disease that leads to tooth-supporting tissues destruction, is one of the most prevalent diseases in the world, and it constitutes the foremost reason of tooth loss in the elderly (Tachi et al., 2003). It results from an imbalance between the levels of potentially pathogenic bacteria in the subgingival microbiota and the immunological potential of the host, which can be modified by several risk factors, including demographic, behavioral, environmental, and systemic aspects (Nunn, 2003). Furthermore, genetic factors may play a role in determining the host's immune response to infection and could account for significant variation in the distribution, severity, and extension of the disease (Nunn, 2003, Zhang, Sun et al., 2011; Laine, Crielaard, & Loos, 2012).

It has been recently demonstrated that one of the factors of innate immune response that might influence the activity of the CP is the activation of the nod-like receptor pyrin domain containing 3 (NLRP3)/apoptosis-associated speck-like protein containing a Caspase recruitment domain (ASC)/Caspase-1 multiprotein complex known as NLRP3 inflammasome in response to various bacterial, physical, and chemical agents (Bostanci et al., 2009, Belibasakis and Johansson, 2012; Belibasakis, Guggenheim, & Bostanci, 2013; Park et al., 2014). The formation of the inflammasome leads to the auto-proteolytic maturation of caspase-1, which subsequently results in maturation and extracellular release of the pro-inflammatory cytokines interleukin (IL)-1β and IL-18 (Dinarello, 2002, Sahdo et al., 2013), that have been associated with inflammatory periodontal tissue destruction (Orozco, Gemmell, Bickel, & Seymour, 2006; Tobón-Arroyave et al., 2008), possibly through a mechanism known as pyroptosis, a form of cell death induced by bacterial pathogens (Strowig, Henao-Mejia, Elinav, & Flavell, 2012), being more potent in its action IL-1β (Dinarello, 1998). Known biological effects of IL-1β include stimulation of T-lymphocytes and cytokine production, proliferation of B-lymphocytes and antibody production (Chiplunkar, Langhorne, & Kaufmann, 1986), fibroblast proliferation, stimulation of prostaglandin E2 (PGE2) release by monocytes and fibroblasts, and release of metalloproteinases that degrade extracellular matrix proteins (Dewhirst, Stashenko, Mole, & Tsurumachi, 1985). IL-1β also promotes osteoclast formation and bone resorption (Gowen & Mundy, 1986), and it affects neutrophil chemotaxis and activation (Westmacott, Wadsworth, & Bloxham, 1987) and endothelial cell function (Goutoudi, Diza, & Arvanitidou, 2004).

Increasing evidence suggests that gain-of-function mutations in the NLRP3 gene lead to a constitutive activation of NLRP3 protein, resulting in an uncontrolled production of IL-1β (Sahdo et al., 2013). Alternatively, it has been postulated that the NLRP3 transcriptional activity can be modulated by single-nucleotide polymorphisms (SNPs) of the NLRP3 gene located on chromosome 1q44. A single base substitution variant (rs4612666) located at intron 7 where a change T > C occurs, is significantly associated with susceptibility to several inflammatory diseases including Crohn disease, psoriatic juvenile idiopathic arthritis, Behçet's syndrome, and rheumatoid arthritis (Day et al., 2008, Villani et al., 2009, Zhang, Zeng et al., 2011, Yüksel et al., 2014, Mathews et al., 2014). However, the association of NLRP3 (rs4612666) SNP with CP has not been described. In addition to the former, the production of IL-1β might be also synchronously modulated by the presence of a functional SNP at the position +3954 of the IL-1B gene (rs1143634) located in the exon 5 of the chromosome 2q13, where a transition between C and T occurs (Pociot, Mølvig, Wogensen, Worsaae, & Nerup, 1992; Nothwang et al., 1997). Nevertheless, the results of association of IL-1B +3954 SNP regarding chronic periodontitis have been conflictive, because while some researchers have found no evidence for association between IL-1B +3954 SNP and CP (Trevilatto, de Souza Pardo, Scarel-Caminaga, Alvim-Pereira, & Alvim-Pereira, 2011; Boukortt et al., 2015, Yücel et al., 2013), others authors have showed a strong association of the CT genotype with disease status in different populations (Shete, Joseph, Vijayan, Srinivas, & Banerjee, 2010; Masamatti, Kumar, Baron, Mehta, & Bhat, 2012; Amirisetty et al., 2015). Therefore, this study aimed to identify if a possible association between NLRP3 (rs4612666) and IL-1B (rs1143634) SNPs may be implicated in the etiopathogenesis of CP in a Colombian population.

Section snippets

Study design, subject recruitment, and inclusion/exclusion criteria

This cross-sectional, observational, analytic study was conducted with a convenience non-probabilistic sample of 205 volunteers from the population of individuals that sought treatment or consultation at the Graduate Periodontics as well as Adult Dental Health Clinics of the University of Antioquia in Medellín (Colombia). The study protocol conformed to the ethical guidelines of the Helsinki Declaration and followed the ethical policy of the Institutional Research Ethics Board. Subsequently,

Post-hoc genetic power calculation and reproducibility of findings

Genetic power calculations suggested that the sample of 124 patients with untreated CP and 81 periodontally HC would provide >93% power to reject the null hypothesis of no association at P < 0.01 when a genotypic relative risk was ≥1.8. Intra-observer reproducibility was excellent for PD (ICC = 0.981 and 0.938 respectively, P < 0.010), and CAL (ICC = 0.978 and 0.999 respectively, P < 0.001) scores in each series of measures recorded per subject by the same examiner. Likewise, inter-observer

Discussion

Over recent decades, immunogenetic studies have provided insight into the individual differences in the pathogenesis of CP and several recent publications have showed that susceptibility to dysbiotic microbial communities with potential for destructive inflammation is based on host genetic factors that may predispose to or protect from disease (Laine et al., 2012, Divaris et al., 2013; Stabholz, Soskolne, & Shapira, 2010; Hajishengallis, 2015). Hence, dysbiosis alone may not necessarily

Acknowledgment

This study has been fully supported by a grant of the Research Development Committee of the University of Antioquia (CODI-Code 2014-481).

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