Elsevier

Experimental Gerontology

Volume 83, October 2016, Pages 56-62
Experimental Gerontology

TNFA gene variants related to the inflammatory status and its association with cellular aging: From the CORDIOPREV study

https://doi.org/10.1016/j.exger.2016.07.015Get rights and content

Highlights

  • TNFA gene variants and cellular aging are linked by oxidative stress-mediated telomere-attrition.

  • TNFA gene variants modulate the inflammatory status and its association with cellular aging.

  • Results support the implication of inflammation-related genes in cellular aging.

  • TNFA gene variants influence telomere length and plasma levels of hs-CRP.

Abstract

Background

Several single nucleotide polymorphisms have been proposed as potential predictors of the development of age-related diseases.

Objective

To explore whether Tumor Necrosis Factor Alpha (TNFA) gene variants were associated with inflammatory status, thus facilitating the rate of telomere shortening and its relation to cellular aging in a population with established cardiovascular disease from the CORDIOPREV study (NCT00924937).

Materials and methods

SNPs (rs1800629 and rs1799964) located at the TNFA gene were genotyped by OpenArray platform in 840 subjects with established cardiovascular disease. Relative telomere length was determined by real time PCR and plasma levels of C-reactive protein by ELISA. In a subgroup of 90 subjects, the gene expression profiles of TNFA, IKKβ, p47phox, p40phox, p22phox and gp91phox were determined by qRT-PCR.

Results

GG subjects for the SNP rs1800629 at the TNFA gene showed shorter relative telomere length and higher plasma levels of hs-CRP than A-allele subjects (p < 0.05). Consistent with these findings, the expression of pro-inflammatory (TNFA) and pro-oxidant (p47phox and the gp91phox) genes was higher in GG subjects than A allele subjects (p < 0.05).

Conclusion

Subjects carrying the GG genotype for the SNP rs1800629 at the TNFA gene show a greater activation of the proinflammatory status than A-allele carriers, which is related to ROS formation. These ROS could induce DNA damage especially in the telomeric sequence, by decreasing the telomere length and inducing cellular aging. This effect may also increase the risk of the development of age-related diseases.

Introduction

Aging is associated with cellular senescence, which is characterized by irreversible cell cycle arrest and dramatic changes in cell morphology and functionality (Colavitti and Finkel, 2005, Collado et al., 2007, Correia-Melo et al., 2014). One of the main contributors to cell cycle arrest is mediated by autocrine signaling involving the secretion of growth factors, inflammatory and immune-modulatory cytokines and chemokines (Coppe et al., 2008). Among the proinflammatory cytokines, overexpression of Tumor Necrosis Factor Alpha (TNFA) has been implicated in a number of pathological conditions related to chronic inflammation and aging. In particular, TNFA induces prolonged growth arrest, decreased telomerase activity and telomeric disruptions (shortening, losses and fusions) (Beyne-Rauzy et al., 2004). Moreover, chronic inflammation exerts its effects through mechanisms that include excessive production of free radicals and depletion of antioxidants, thus leading to excess oxidative stress. The latter has been implicated in the pathogenesis of several age-related diseases such as diabetes and cardiovascular disease (CVD) (Ceriello and Motz, 2004, Durackova, 2010). Reactive oxygen species (ROS) are likely to be involved in both the induction and stabilization of cellular senescence and numerous reports point to links between oxidative damage and the aging process (Hamilton et al., 2001). ROS is also associated with a gradual loss of DNA at the ends of chromosomes and eventual telomere dysfunction (Erusalimsky and Kurz, 2005) that contributes to cellular senescence (Khan et al., 2012). The shortening rate can be accelerated by several factors, including age (Armanios, 2013), phenotype (Masi et al., 2014), oxidative stress (von Zglinicki, 2000) and endothelial dysfunction (Gonzalez-Guardia et al., n.d.). For instance, when cells are exposed to high levels of oxidative stress, the amount of telomere attrition per cell division significantly increases (Jena, 2012). Its study is therefore crucial for understanding mechanisms associated with the development of age-related diseases (Armanios, 2013).

Both aging and age-related diseases have a significant genetic component (Johnson et al., 2015, Richardson and Schadt, 2014), and several studies have demonstrated associations between single nucleotide polymorphism (SNPs) and aging-associated diseases including CVD (Angelakopoulou et al., n.d, Corsetti et al., n.d, Vasto et al., 2007). Some of these studies have pointed out the significant relationship between the TNFA locus and CVD (Cui et al., 2012, Mellick, 2007), in particular, it has been demonstrated that two SNPs transition substitution-type rs1800629, NM_000594.3:c, − 488A > G, also known as − 308G > A, and rs1799964, NM_000595.3:c, − 838C > T, located in the TNFA gene, are associated with CVD development (Chu et al., 2012, Hernandez-Diaz et al., 2015, Rodriguez-Rodriguez et al., 2011, Sandoval-Pinto et al., 2015). Moreover, the development and progression of CVD is related to cellular senescence, and proinflammatory cytokines may be the nexus for both processes. However, the mechanisms are still unclear and genetic tools may be the key to unravelling the mechanisms driving the aging process and its inter-individual differences observed in the population.

Based on this concept and the existing evidence, we explored whether genetic variants at the TNFA gene were associated with the inflammatory status, thus mediating telomere length and its relation to cellular aging in a population with established CVD.

Section snippets

Population

The current work was conducted within the framework of the CORDIOPREV study. The CORDIOPREV study is an ongoing prospective, randomized, opened, controlled trial including 1002 patients with coronary heart disease (CHD), who had their last coronary event more than six months before enrolment. Patients were randomized in two different dietary models (Mediterranean and low-fat) over a period of five years, in addition to conventional treatment for CHD. Patients were recruited from November 2009

Characteristics of the study participants classified according the TNFA SNPs genotype

The demographic, anthropometric and biochemical characteristics according to the TNFA SNPs in each genotype (rs1800629 and rs1799964) are presented in Table 1. The genotype distributions did not deviate from the Hardy-Weinberg expectations. We compared the allelic frequencies observed for both TNFA SNPs with the 1000genomes database. The minor allele frequency for the SNP rs1800629 was A = 0.106 (1000genomes: A = 0.144) and for the SNP rs1799964 was C = 0.271 (1000genomes: C = 0.177). The genotypic

Discussion

Our results show that carriers of the GG genotype for the SNP rs1800629 at the TNFA gen have lower RTL and higher hs-CRP plasma levels than A allele carriers. Moreover, an association between telomere length and inflammation was also observed based on the increased expression levels of TNFA and IKKβ genes in GG subjects. These results support the implication of inflammation-related genes in the rate of cellular aging.

Inflammation plays an important role in the pathogenesis of atherosclerosis

Authors' contributions

The author's responsibilities were as follows: OARZ and PP-M conceived and designed the experiments; AC, FG-D, JFA-D, JD-L, JL-M, and PP-M participated in the recruitment and carried out the clinical and nutritional control of the volunteers; OARZ, AC, BL-P, and CC-T, performed the experiments and collected the data; OARZ, CA, BL-P, RJ-L, CH-M, EMY-S and PP-M analyzed and interpreted the data; OARZ and PP-M drafted the manuscript; FR-C was responsible for the management of samples and

Conflict of interest and funding disclosure

None of the authors has any conflict of interest that could affect the performance of the work or the interpretation of the data.

Acknowledgements

We would like to thank the EASP (Escuela Andaluza de Salud Publica), Granada, Spain, which performed the randomization process for this study. The CORIOPREV study is supported by the Fundación Patrimonio Comunal Olivarero, Junta de Andalucía (Consejería de Salud, Consejería de Agricultura y Pesca, Consejería de Innovación, Ciencia y Empresa), Diputaciones de Jaén y Córdoba, Centro de Excelencia en Investigación sobre Aceite de Oliva y Salud and Ministerio de Medio Ambiente, Medio Rural y

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