The rs2066808 Polymorphism Located Near the IL-23A Gene Is Associated with Premature Coronary Artery Disease in Mexican Population (GEA Study)

Abstract

Interleukin-23 (IL-23) has been associated with atherosclerosis in both humans and animal models with contradictory results. This cytokine is conformed by an α p19 (encoded by IL-23A gene) and a β p40 subunit (encoded by IL-12B gene). The aim of this study was to evaluate the association of two polymorphisms located within (rs11171806) or near (rs2066808) of the IL-23A gene with the presence of premature coronary artery disease (CAD) and with cardiometabolic parameters. The rs2066808 and rs11171806 polymorphisms were determined in 2249 Mexican individuals (1160 with premature CAD and 1089 healthy controls). Under recessive and codominant 2 models, adjusted by confounding variables, the rs2066808 polymorphism could increase the genetic risk of premature CAD (odds ratio [OR] = 4.567, 95% confidence interval [CI]: 1.03–20.24, P_recessive = 0.046 and OR = 4.606, 95% CI: 1.039–20.43, P_codominant2 = 0.044). The association of the polymorphisms with cardiovascular risk factors was evaluated separately in premature CAD patients and healthy controls. In patients, the rs2066808 polymorphism could decrease the genetic risk of hyperinsulinemia, insulin resistance, and hypoalphalipoproteinemia, and increase the genetic risk of hyperuricemia, whereas the rs11171806 polymorphism could decrease the genetic risk of hyperinsulinemia and insulin resistance. In healthy controls, the rs11171806 polymorphism could decrease the genetic risk of hyperinsulinemia. These findings suggest that the rs2066808 polymorphism located near the IL-23A gene could increase the genetic risk of premature CAD and both studied polymorphisms could be associated with some cardiometabolic parameters in premature CAD patients and in healthy controls.

Introduction

Atherosclerosis is a chronic, progressive, and multifactorial disease modulated by genetic and environmental factors. An important complication of atherosclerosis is coronary artery disease (CAD). This disease is considered the most common cause of death in industrialized countries, accounting for up to 40% of all deaths (Lopez et al., 2006). An increasing number of observations have shown that inflammation plays an important role in the pathogenesis of atherosclerosis and its complications. An imbalance of anti- and proinflammatory cytokines in the development and progression of atherosclerosis has been described in animal models (Frostegård et al., 1999; Tedgui and Mallat, 2006).

Interleukin 23 (IL-23), a member of the interleukin 12 family, is a heterodimeric cytokine, conformed by an α p19 and a β p40 subunit (Oppmann et al., 2000). This cytokine is produced mainly by macrophages and dendritic cells after toll-like receptor activation (Aggarwal et al., 2003) and plays a central role in the inflammatory process. IL-23 stimulates the proliferation of T memory cells, the development of proinflammatory Th17 cells, and the differentiation of Th1 cells (Aggarwal et al., 2003). IL-23 has been associated with atherosclerosis in both humans and animal models with contradictory results. Lower IL-23 gene expression was observed in unstimulated peripheral blood lymphocytes of patients with CAD when compared with healthy controls (Khojasteh-Fard et al., 2012). However, in another study, high IL-23 levels were detected in patients with carotid atherosclerosis when compared with healthy controls, and these levels were associated with disease progression and increased mortality (Abbas et al., 2015). In contrast, in an IL-23p19−/− mice model with myocardial infarction, the IL-23 deficiency produces myocardial inflammation, lower cardiac fibroblast activation, ventricular rupture, and increased mortality (Savvatis et al., 2014).

Beta subunit p40 is encoded by IL-12B gene on chromosome 5. Information regarding the association of polymorphisms present in the IL-12B gene with CAD is scarce. A Japanese study in a small cohort failed to show an association between rs3212227 IL-12B polymorphism and the presence or severity of CAD diagnosed by angiography (Momiyama et al., 2005). These results agree with those reported by Mangino et al. (2008) in a British cohort. In contrast, the α subunit p19 is encoded by the IL-23A gene located on 12q13.3 region. Polymorphisms in this gene have been associated with the presence of several diseases (Catanoso et al., 2013; Costa et al., 2013; Li et al., 2016); however, no studies have evaluated the association of the IL23A polymorphisms with the development of cardiovascular disease. Considering this, the aim of this study was to evaluate the association of two polymorphisms located within (rs11171806) or near (rs2066808) of the IL-23A gene with the presence of CAD in a cohort of Mexican individuals.

Materials and Methods

Subjects

The study included 2249 Mexican individuals (1160 with premature CAD and 1089 healthy controls) belonging to the Genetics of Atherosclerosis (GEA) Mexican study. The premature CAD patients were diagnosed by history of myocardial infarction, angioplasty, revascularization surgery, or coronary stenosis >50% on angiography. Premature CAD was considered when the diagnosis was made before age 55 in men and before age 65 in women. The individuals included in the control group were recruited from blood bank and through brochures posted in social service centers. All were apparently healthy asymptomatic individuals without family history of premature CAD. To obtain family medical history, demographic information, history of nutritional habits, physical activity, alcohol consumption, and pharmacological treatment in patients and healthy controls, standardized questionnaires were applied. Anthropometric, biochemical, metabolic, and cardiovascular risk factors were evaluated in both premature CAD cases and controls and defined as previously described (Posadas-Sánchez et al., 2014, 2017a; Medina-Urrutia et al., 2015). Considering that the Mexican population is an admixture of Caucasian, Amerindian, and African genes, these components were evaluated previously using 265 ancestry markers (Posadas-Sánchez et al., 2017b). These results show that global ancestry was not significantly different between premature CAD patients and healthy controls (55.8% vs. 54.0% Amerindian ancestry, 34.3% vs. 35.8% Caucasian, and 9.8% vs. 10.1% African mean ancestry for patients and controls, respectively, p > 0.05). This suggests that population stratification was not a bias or confounding factor in this study.

The GEA study was approved by the Bioethics Committee of the Instituto Nacional de Cardiología Ignacio Chávez (INCICH, number 19-1104), and aligned to the Helsinki Declaration. All participants provided informed consent.

Computed axial tomography study

Coronary artery calcification (CAC) score (Mautner et al., 1994), total, subcutaneous, and visceral abdominal fat areas (Kvist et al., 1988), and hepatic to splenic attenuation ratio (Longo et al., 1993) were determined after a computed tomography of the chest and abdomen performed using a 64-channel multidetector helical computed tomography system (Somatom Sensation, Siemens). The GEA study originally included 1500 healthy controls; yet, after the computed tomography, the CAC score of 411 subjects was >0. They were thus considered as individuals with subclinical atherosclerosis and were not included in the analysis. The final healthy control group included 1089 individuals with a CAC score of 0.

Genetic analysis

The rs2066808 (C__12055277_10) and rs11171806 (C__25985467_10) polymorphisms were determined using 5′ exonuclease TaqMan assays. The determinations were made with an ABI Prism 7900HT Fast Real-Time PCR system, according to manufacturer's instructions (Applied Biosystems, Foster City, CA). To corroborate the adequate assignment of the genotypes in the TaqMan assays, we randomly select and repeat 10% of the samples. These samples were 100% concordant in two independent assays.

Statistical analysis

The analysis was made using the SPSS version 15.0 statistical package (SPSS, Chicago, IL). Means, medians, interquartile ranges, and frequencies were calculated as the case may be. Continuous and categorical variables were analyzed by Student's t test, Mann–Whitney U-test, Kruskal–Wallis, and chi-square as appropriate. The association of the polymorphisms with premature CAD, metabolic parameters, and cardiovascular risk factors was analyzed by logistic regression under the following inheritance models: additive (major allele homozygotes vs. heterozygotes vs. minor allele homozygotes), codominant 1 (major allele homozygotes vs. heterozygotes), codominant 2 (major allele homozygotes vs. minor allele homozygotes), dominant (major allele homozygotes vs. heterozygotes+minor allele homozygotes), overdominant (heterozygotes vs. major allele homozygotes+minor allele homozygotes), and recessive (major allele homozygotes+heterozygotes vs. minor allele homozygotes). In the analysis of association with premature CAD, the models were adjusted by age, gender, body mass index (BMI), current smoking, low- and high-density lipoprotein cholesterol (LDL-C and HDL-C), and homeostasis model assessment-insulin resistance (HOMA-IR), whereas in the associations with metabolic parameters and cardiovascular risk factors, the models were adjusted by age, gender and BMI. p < 0.05 values were considered statistically significant. Genotype frequencies did not deviate from Hardy–Weinberg equilibrium in any case (p > 0.05). The haplotypes were constructed and analyzed using the Haploview version 4:1 (Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA).

Results

Demographic, biochemical, and cardiovascular risk factors

Demographic, biochemical, and cardiovascular risk factors of the studied groups are given in Table 1. All the variables were increased in the patient group except the total cholesterol <200 mg/dL, the LDL-C ≥130 mg/dL, non-HDL-C >160 mg/dL, smoking habit, and high sensitivity C-reactive protein ≥3 mg/L that were decreased in this group compared with healthy controls. The decrease of these variables in patients after diagnosis could be due to patients received statin treatment and in addition modified their life styles by medical recommendation.

Table 1.

Demographic, Biochemical, and Cardiovascular Risk Factors Prevalence of the Studied Population

	Controls (n = 1089)	Premature CAD (n = 1160)	p
Age (years)	51 ± 9	54 ± 8	<0.001
Gender (% male)	41.0	81.0	<0.001
BMI (kg/m²)	27.9 [25.4–30.8]	28.3 [25.9–31.1]	0.004
Waist circumference (cm)	94 ± 11	98 ± 10	<0.001
Systolic blood pressure (mmHg)	112 [104–123]	116 [106–127]	<0.001
Diastolic blood pressure (mmHg)	70.5 [65.0–76.0]	71.0 [65.5–77.5]	0.011
Visceral adipose fat (cm²)	139 [105–181]	168 [129–215]	<0.001
Alanine aminotransferase (IU/L)	24[18–34]	26 [19–36]	0.025
Aspartate aminotransferase (IU/L)	25 [21–30]	26 [22–31]	0.001
Total cholesterol <200 mg/dL (%)	36.4	20.3	<0.001
LDL-C ≥130 mg/dL (%)	29.6	16.1	<0.001
Hipoalphalipoproteinemia (%)	52.1	67.1	<0.001
Non-HDL-C >160 mg/dL (%)	27.9	19.5	<0.001
Hypertriglyceridemia (%)	47.3	56.2	<0.001
Obesity (%)	30.3	35.1	0.009
Abdominal obesity (%)	81.2	83.7	0.067
Type 2 diabetes mellitus (%)	9.6	35.5	<0.001
Hyperinsulinemia (%)	52.2	66.1	<0.001
Insulin resistance (%)	54.2	72.8	<0.001
Hypertension (%)	18.9	68.1	<0.001
High visceral abdominal fat (%)	54.7	64.7	<0.001
Current smoking (%)	22.6	11.6	<0.001
Hypoadiponectinemia (%)	42.9	57.6	<0.001
High sensitivity CRP ≥3 mg/L (%)	26.7	20.5	0.002
Hyperuricemia (%)	20.5	36.1	<0.001

Data are shown as mean ± standard deviation, median [interquartile range], or percentage. Comparisons were made using Student's t test or Mann–Whitney U-test, as appropriate, for continuous variables, and by chi-square analysis for categorical variables.

BMI, body mass index; CAD, coronary artery disease; CRP, C-reactive protein; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol.

Association of the polymorphisms with premature CAD

The distribution of the studied polymorphisms in premature CAD patients and healthy controls is given in Table 2. The distribution of the rs11171806 polymorphism was similar in patients and healthy controls. In contrast, under the recessive and codominant 2 models, the rs2066808 polymorphism could increase the genetic risk of premature CAD (odds ratio [OR] = 4.567, 95% confidence interval [CI]: 1.03–20.24, P_recessive = 0.046 and OR = 4.606, 95% CI: 1.039–20.43, P_codominant2 = 0.044). The models were adjusted by age, gender, BMI, current smoking, LDL-C, HDL-C, and HOMA-IR. The statistical power to detect association of the rs2066808 polymorphism with premature CAD was >90% as estimated with QUANTO software [biostats.usc.edu/Quanto.html].

Table 2.

Association Between Interleukin-23A Polymorphisms and Premature Coronary Disease

Polymorphism	Genotype frequency			MAF	Model	OR [95% CI]	p
rs2066808	AA	AG	GG		Additive	1.179 [0.906–1.535]	0.221
Control (n = 1089)	0.851	0.146	0.003	0.076	Dominant	1.125 [0.848–1.491]	0.414
					Recessive	4.567 [1.030–20.24]	0.046
CAD (n = 1160)	0.848	0.142	0.009	0.081	Overdominant	1.049 [0.787–1.397]	0.787
					Codominant 1	1.060 [0.796–1.413]	0.689
					Codominant 2	4.606 [1.039–20.43]	0.044
rs11171806	GG	GA	AA		Additive	1.163 [0.848–1.595]	0.349
Control (n = 1089)	0.896	0.102	0.002	0.053	Dominant	1.176 [0.846–1.635]	0.334
					Recessive	1.070 [0.175–6.539]	0.942
CAD (n = 1160)	0.896	0.101	0.003	0.053	Overdominant	1.179 [0.844–1.645]	0.334
					Codominant 1	1.179 [0.845–1.646]	0.333
					Codominant 2	1.089 [0.178–6.666]	0.926

Models were adjusted for age, gender, BMI, current smoking, LDL-C and HDL-C, and HOMA-IR.

Bold numbers indicate significant associations.

CI, confidence interval; HOMA-IR, homeostasis model assessment-insulin resistance; MAF, minor allele frequency; OR, odds ratio.

Association of the polymorphisms with cardiovascular risk factors

The association of the polymorphisms with cardiovascular risk factors was evaluated separately in premature CAD patients and healthy controls. In patients (Table 3), the rs2066808 polymorphism could decrease the genetic risk of hyperinsulinemia (OR = 0.271, P_recessive = 0.048; OR = 0.268, P_codominant2 = 0.046), insulin resistance (OR = 0.276, P_recessive = 0.043; OR = 0.270, P_codominant2 = 0.040), and hypoalphalipoproteinemia (OR = 0.269, P_recessive = 0.042; OR = 0.274, P_codominant2 = 0.045), and increase the risk of hyperuricemia (OR = 1.459, P_additive = 0.015; OR = 1.486, P_dominant = 0.021; OR = 1.412, P_overdominant = 0.045; OR = 1.436, P_codominant1 = 0.039). In the same way, the rs11171806 polymorphism could decrease the genetic risk of developing hyperinsulinemia (OR = 0.677, P_additive = 0.043; OR = 0.645, P_dominant = 0.031; OR = 0.632, P_{over-dominant} = 0.026; OR 0.632, P_codominant1 = 0.026) and insulin resistance (OR = 0.659, P_additive = 0.037; OR = 0.632, P_dominant = 0.029; OR = 0.625, P_overdominant = 0.027; OR = 0.625, P_codominant1 = 0.027). In contrast, in healthy controls (Table 4), the rs11171806 polymorphism could decrease the risk of hyperinsulinemia (OR = 0.645, P_additive = 0.049).

Table 3.

Association Between Interleukin-23A Polymorphisms and Cardiometabolic Abnormalities in Coronary Artery Disease Patients

	Genotype frequency			MAF	Model	OR [95% CI]	P
rs2066808	AA	AG	GG
Hyperinsulinemia
No (n = 393)	0.835	0.147	0.018	0.092	Recessive	0.271 [0.075–0.987]	0.048
Yes (n = 767)	0.856	0.139	0.005	0.075	Codominant 2	0.268 [0.074–0.976]	0.046
Insulin resistance
No (n = 316)	0.827	0.154	0.019	0.097	Recessive	0.276 [0.079–0.963]	0.043
Yes (n = 844)	0.857	0.137	0.006	0.075	Codominant 2	0.270 [0.077–0.942]	0.040
Hypoalphalipoproteinemia
No (n = 382)	0.851	0.131	0.018	0.084	Recessive	0.269 [0.076–0.951]	0.042
Yes (n = 778)	0.847	0.148	0.005	0.079	Codominant 2	0.274 [0.077–0.971]	0.045
Hyperuricemia
No (n = 741)	0.867	0.126	0.007	0.070	Additive	1.459 [1.076–1.979]	0.015
Yes (n = 419)	0.817	0.168	0.014	0.099	Dominant	1.482 [1.061–2.070]	0.021
					Overdominant	1.412 [1.008–2.003]	0.045
					Codominant 1	1.436 [1.018–2.025]	0.039
rs11171806	GG	GA	AA
Hyperinsulinemia
No (n = 393)	0.869	0.129	0.003	0.066	Additive	0.677 [0.464–0.988]	0.043
Yes (n = 767)	0.909	0.087	0.004	0.048	Dominant	0.645 [0.433–0.960]	0.031
					Overdominant	0.632 [0.422–0.946]	0.026
					Codominant 1	0.632 [0.422–0.947]	0.026
Insulin resistance
No (n = 316)	0.862	0.135	0.003	0.071	Additive	0.659 [0.446–0.974]	0.037
Yes (n = 844)	0.908	0.089	0.004	0.048	Dominant	0.632 [0.419–0.954]	0.029
					Overdominant	0.625 [0.412–0.949]	0.027
					Codominant 1	0.625 [0.412–0.948]	0.027

Models were adjusted for age, gender, and BMI.

All significant models are shown.

Table 4.

Association Between Interleukin-23A Polymorphisms and Cardiometabolic Abnormalities in Control Subjects

	Genotype frequency			MAF	Model	OR [CI 95%]	P
Hyperinsulinemia
rs11171806	GG	GA	AA
No (n = 519)	0.882	0.114	0.004	0.061	Additive	0.645 [0.418–0.997]	0.049
Yes (n = 570)	0.908	0.092	0.000	0.046

Models were adjusted for age, gender, and BMI.

Haplotype analysis

The polymorphisms were in high linkage disequilibrium (D′ = 0.98, r ² = 0.64); however, a similar distribution of the haplotypes constructed was observed in patients and healthy controls (data not shown).

Discussion

In a group of 2249 Mexican individuals (1160 with premature CAD and 1089 controls), we describe that the rs2066808 polymorphism located near the IL-23A gene could increase the genetic risk of developing premature CAD. In premature CAD patients, the rs2066808 polymorphism could decrease the risk of hyperinsulinemia, insulin resistance, and hypoalphalipoproteinemia, and increase the risk of hyperuricemia, whereas the rs11171806 could decrease the risk of hyperinsulinemia and insulin resistance. In contrast, in controls, the rs2066808 could decrease the risk of hyperinsulinemia. As can be seen, the association of the polymorphisms with cardiometabolic parameters was different in premature CAD patients and healthy controls, with the most important effect in patients. In this group, both polymorphisms could decrease the risk of hyperinsulinemia and insulin resistance, two related characteristics. Thus, patients with the rs2066808G and rs11171806A alleles have low insulin resistance, and in consequence they do not need the insulin resistance/compensatory hyperinsulinemia, having low risk for this condition.

IL-23 is a heterodimeric cytokine with important effects in the immune system through its participation in the inflammatory process. IL-23 has been associated with the development of several inflammatory disorders such as rheumatoid arthritis (Rong et al., 2012), systemic lupus erythematosus (Xia et al., 2015), psoriasis (Mudigonda et al., 2012), ulcerative colitis (Mohammadi et al., 2011), and inflammatory bowel disease (Yen et al., 2006). Its role in the development of atherosclerosis has been studied in animal models and humans with contradictories results (Khojasteh-Fard et al., 2012; Savvatis et al., 2014; Abbas et al., 2015; Posadas-Sánchez and Vargas-Alarcón, 2018).

It is well know that IL-23 secretion is limited by the tissue and cell-restricted expression of p19 subunit (Oppmann et al., 2000), encoded by the IL-23A gene. Thus, functional genetic variants within the IL-23A gene could have important impact on immune response and could be candidate gene for association studies (Tindall and Hayes, 2010). Association studies of the IL-23A polymorphisms with some diseases have been reported (Catanoso et al., 2013; Costa et al., 2013; Li et al., 2016). The IL-23A rs2066808, rs2371494, and rs11575248 polymorphisms were associated with susceptibility to multiple sclerosis in Chinese Han population (Li et al., 2016), whereas in Brazilians, the GG haplotype of rs11171806 and rs2066808 polymorphisms decreases the risk of type 1A diabetes mellitus (Costa et al., 2013). In contrast, in German and Chinese cohorts, the rs2066808 polymorphism was associated with susceptibility to psoriasis (Nair et al., 2009; Chen et al., 2011), whereas in Spaniards, this polymorphism was associated with psoriatic arthritis (Eiris et al., 2012). Two independent cohorts of Romanians and Caucasians (British and Ireland) confirm these results (Bowes et al., 2011; Popa et al., 2013). Unfortunately, at the present time, there are no studies that have evaluated the association of polymorphism within or near the IL23A gene with the development of cardiovascular disease. In our study, we confirm the important role of the rs2066808 polymorphism now in a cardiovascular disease, the premature CAD.

IL-23 is a cytokine that is included in the IL-12 family together with IL-12, IL-27, and IL-35 cytokines. This family consists of a single group of α/β heterodimeric cytokines composed of one of three alpha chains (p19 encode by IL-23A, p28 encode by IL-27p28, or p35 encode by IL-12A) and one of two beta chains [p40 encoded by IL-12B gene or Epstein–Barr virus-induced gene 3 (EBI3) encoded by EBI3] (Vignali et al., 2012). Previously, in the same cohort reported in this study, we analyzed the association of the polymorphisms of the IL-27p28, IL-12A, and EBI3 genes with premature CAD (Posadas-Sánchez et al., 2017b, 2017c). With the analysis of IL-23A polymorphisms, we complete the study of polymorphisms located in genes that encode the subunits that constitute the IL-12 family cytokine in our cohort of patients with premature CAD.

In contrast, the association of the rs2066808 polymorphism located near the IL-23A gene with premature CAD reported in this study is in accordance with the proinflammatory effects of this cytokine. This polymorphism is located in an intronic region of the STAT2 (signal transducer and activator of transcription) gene and could alter RNA splicing by disrupting splice site enhancers or silencers, resulting in an isoform of this molecule with altered function. STAT2 is a transcription activator related with the signal transduction pathway of type 1 interferon (Platanias, 2005) and with immunosuppressive function (Yi et al., 2012). The potential role of STAT2 in atherosclerosis has been previously described in an animal model (Lagor et al., 2014). In this study, it was reported that ApoF-deficient mice under Western diet are protected from atherosclerosis and is suggested that this protection is related with a reduced expression of STAT2 gene. STAT together with IRF (interferon regulatory factor) plays an important role in fundamental cellular process such as development, apoptosis, cell growth and differentiation, immune response, and inflammation (Levy and Darnell, 2002), all of them process involved in the developing atherosclerosis.

It is important to consider that our control group included only individuals without CAC evaluated by tomography. The subjects with calcium score >0 were considered as individuals with subclinical atherosclerosis and were not included as control individuals. The proportion of Caucasian, Native American, and African ancestries was similar in patients and controls, so population stratification was ruled out as a potential confounding factor. An important number of clinical, biochemical, and tomographic variables were collected in the studied groups that allowed adjustment of the analyses for a large number of potential confounders. Despite all of these strengths of our study, some limitations should also be considered. First, we only determined two polymorphisms that were previously associated with inflammatory diseases in other populations. Second, due to the transversal character of the study, conclusions on causality cannot be made. Third, the functional effect of the polymorphisms was not defined experimentally. Finally, the statistical power to detect association of the rs11171806 polymorphism with premature CAD was < 40%, so, the result reported in our study of this polymorphism must be taken with care.

In summary, our data suggest that the rs2066808 polymorphism located near the IL-23A gene could increase the genetic risk of premature CAD. Both polymorphisms studied were associated with some cardiometabolic parameters principally in the patient group. Associations with hyperinsulinemia, insulin resistance, hypoalphalipoproteinemia, and hyperuricemia were reported.

Footnotes

Acknowledgments

This study was supported by a grant from the Consejo Nacional de Ciencia y Tecnología (Fronteras de la Ciencia No. 1958), Mexico City, Mexico. C.V.-V. is a doctoral student from Programa de Doctorado en Ciencias Biomédicas, Universidad Nacional Autónoma de México (UNAM). The authors are grateful to the study participants. Institutional Review Board approval was obtained for all sample collections. The authors are grateful to Marva Arellano-Gonzalez and Silvestre Ramírez-Fuentes for their technical assistance.

Authors' Contributions

C.V.-V. and G.V.-A. were responsible for the conception and design of the study. R.P.-S. and N.P.-H. contributed to the analysis and interpretation of data. J.M.F., J.M.R.-P., and G.C.-S. participated in the collection of samples and carried out the experiments. Drafting and revision of the article was handled by G.V.-A., C.V.-V., and R.P.-S.

Disclosure Statement

No competing financial interests exist.

References

Abbas

, Gregersen

, Holm

, Daissormont

, Bjerkeli

, Krohg-Sørensen

, et al. (2015). Interleukin 23 levels are increased in carotid atherosclerosis. Possible role for the interleukin 23/interleukin 17 axis. Stroke, 46, 793–799.

Aggarwal

, Ghilardi

, Xie

M.-H.

, de Sauvage

F.J.

, and Gurney

A.L.

(2003). Interleukin-23 promotes a distinct CD4 T cell activation state characterized by the production of interleukin-17. J Biol Chem, 278, 1910–1914.

Bowes

, Orozco

, Flynn

, Ho

, Brier

, Marzo-Ortega

, et al. (2011). Confirmation of TNIP1 and IL23A as susceptibility loci for psoriatic arthritis. Ann Rheum Dis, 70, 1641–1644.

Catanoso

M.G.

, Boiardi

, Macchioni

, Garagnani

, Sazzini

, De Fanti

, et al. (2013). IL-23A, IL-23R, IL-17A and IL-17R polymorphisms in different psoriatic arthritis clinical manifestations in the northern Italian population. Rheumatol Int, 33, 1165–1176.

Chen

, Poon

, Yeung

, Helms

, Pons

, Bowcock

A.M.

, et al. (2011). A genetic risk score combining ten psoriasis risk loci improves disease prediction. PLoS One 6, e19454.

Costa

V.S.

, Santos

A.S.

, Fukui

R.T.

, Mattana

T.C.C.

, Matioli

S.R.

, and Silva

M.E.R.

(2013). Protective effect of interleukin-23A (IL23A) haplotype variants on type 1A diabetes mellitus in a Brazilian population. Cytokine, 62, 327–333.

Eiris

, Santos-Juanes

, Coto-Segura

, Gómez

, Alvarez

, Morales

, et al. (2012). Resequencing of the IL12B gene in psoriasis patients with the rs6887695/rs3212227 risk genotypes. Cytokine, 60, 27–29.

Frostegård

, Ulfgren

A.K.

, Nyberg

, Hedin

, Swedenborg

, Andersson

, et al. (1999). Cytokine expression in advanced human atherosclerotic plaques: dominance of pro-inflammatory (Th1) and macrophage-stimulating cytokines. Atherosclerosis, 145, 33–43.

Khojasteh-Fard

, Abolhalaj

, Amiri

, Zaki

, Taheri

, Qorbani

, et al. (2012). IL-23 gene expression in PBMCs of patients with coronary artery disease. Dis Markers, 33, 289–293.

10.

Kvist

, Chowdhury

, Grangård

, Tylén

, and Sjöström

(1988). Total and visceral adipose-tissue volumes derived from measurements with computed tomography in adult men and women: predictive equations. Am J Clin Nutr, 48, 1351–1361.

11.

Lagor

W.R.

, Fields

D.W.

, Bauer

R.C.

, Crawford

, Abt

M.C.

, Artis

, et al. (2014). Genetic manipulation of the ApoF/Stat2 locus supports an important role for type I interferon signaling in atherosclerosis. Atherosclerosis, 233, 234–241.

12.

Levy

D.E.

, and Darnell

J.E.

(2002). STATs: transcriptional control and biological impact. Nat Rev Mol Cell Biol, 3, 651–662.

13.

F.-F.

, Zhu

X.-D.

, Yan

, Jin

M.-H.

, Yue

, Zhang

, et al. (2016). Characterization of variations in IL23A and IL23R genes: possible roles in multiple sclerosis and other neuroinflammatory demyelinating diseases. Aging (Albany NY), 8, 2734–2746.

14.

Longo

, Ricci

, Masutti

, Vidimari

, Crocé

L.S.

, Bercich

, et al. (1993). Fatty infiltration of the liver. Quantification by 1H localized magnetic resonance spectroscopy and comparison with computed tomography. Invest Radiol, 28, 297–302.

15.

Lopez

A.D.

, Mathers

C.D.

, Ezzati

, Jamison

D.T.

, and Murray

C.J.

(2006). Global and regional burden of disease and risk factors, 2001: systematic analysis of population health data. Lancet, 367, 1747–1757.

16.

Mangino

, Braund

, Singh

, Steeds

, Stevens

, Channer

K.S.

, et al. (2008). Association analysis of IL-12B and IL-23R polymorphisms in myocardial infarction. J Mol Med, 86, 99–103.

17.

Mautner

G.C.

, Mautner

S.L.

, Froehlich

, Feuerstein

I.M.

, Proschan

M.A.

, Roberts

W.C.

, et al. (1994). Coronary artery calcification: assessment with electron beam CT and histomorphometric correlation. Radiology, 192, 619–623.

18.

Medina-Urrutia

, Posadas-Romero

, Posadas-Sánchez

, Jorge-Galarza

, Villarreal-Molina

, González-Salazar

M.D.C.

, et al. (2015). Role of adiponectin and free fatty acids on the association between abdominal visceral fat and insulin resistance. Cardiovasc Diabetol 14, 20.

19.

Mohammadi

, Hayatbakhsh

M.M.

, Zahedi

M.J.

, Jalalpour

M.R.

, and Pakgohar

(2011). Serum interleukin-23 levels in patients with ulcerative colitis. Iran J Immunol, 8, 183–188.

20.

Momiyama

, Ohmori

, Nagano

, Kato

, Taniguchi

, Egashira

, et al. (2005). Polymorphism of the 3′-untranslated region of interleukin-12 p40 gene is not associated with the presence or severity of coronary artery disease. Circ J, 69, 793–797.

21.

Mudigonda

, Mudigonda

, Feneran

A.N.

, Alamdari

H.S.

, Sandoval

, and Feldman

S.R.

(2012). Interleukin-23 and interleukin-17: importance in pathogenesis and therapy of psoriasis. Dermatol Online J 18, 1.

22.

Nair

R.P.

, Duffin

K.C.

, Helms

, Ding

, Stuart

P.E.

, Goldgar

, et al. (2009). Genome-wide scan reveals association of psoriasis with IL-23 and NF-kappaB pathways. Nat Genet, 41, 199–204.

23.

Oppmann

, Lesley

, Blom

, Timans

J.C.

, Xu

, Hunte

, et al. (2000). Novel p19 protein engages IL-12p40 to form a cytokine, IL-23, with biological activities similar as well as distinct from IL-12. Immunity, 13, 715–725.

24.

Platanias

L.C.

(2005). Mechanisms of type-I- and type-II-interferon-mediated signalling. Nat Rev Immunol, 5, 375–386.

25.

Popa

O.M.

, Kriegova

, Popa

, Schneiderova

, Dutescu

M.I.

, Bojinca

, et al. (2013). Association study in Romanians confirms IL23A gene haplotype block rs2066808/rs11171806 as conferring risk to psoriatic arthritis. Cytokine, 63, 67–73.

26.

Posadas-Sánchez

, Angélica

, Arcos

, René, Á., Uribe

, González-salazar

M.C.

, et al. (2014). Asociación del ácido úrico con factores de riesgo cardiovascular y aterosclerosis subclínica en adultos mexicanos. [Association of uric acid with cardiovascular risk factors and subclinical atherosclerosis in Mexican adults]. [in Spanish]. Rev Mex Endocrinol Metab Nutr, 1, 14–21.

27.

Posadas-Sánchez

, López-Uribe

Á.R.Á.R.

, Posadas-Romero

, Pérez-Hernández

, Rodríguez-Pérez

J.M.J

.M.J.M., Ocampo-Arcos

W.A.W

.A., et al. (2017a). Association of the I148M/PNPLA3 (rs738409) polymorphism with premature coronary artery disease, fatty liver, and insulin resistance in type 2 diabetic patients and healthy controls. The GEA study. Immunobiology, 222, 960–966.

28.

Posadas-Sánchez

, Pérez-Hernández

, Angeles-Martínez

, López-Bautista

, Villarreal-Molina

, Rodríguez-Pérez

J.M.

, et al. (2017b). Interleukin 35 polymorphisms are associated with decreased risk of premature coronary artery disease, metabolic parameters, and IL-35 levels: the genetics of atherosclerotic disease (GEA) study. Mediators Inflamm 2017, 6012795.

29.

Posadas-Sánchez

, Pérez-Hernández

, Rodríguez-Pérez

J.M.

, Coral-Vázquez

R.M.

, Roque-Ramírez

, Llorente

, et al. (2017c). Interleukin-27 polymorphisms are associated with premature coronary artery disease and metabolic parameters in the Mexican population: the genetics of atherosclerotic disease (GEA) Mexican study. Oncotarget, 8, 64459–64470.

30.

Posadas-Sánchez

, and Vargas-Alarcón

(2018). Innate immunity in coronary disease. The role of interleukin-12 cytokine family in atherosclerosis. Rev Investig Clin, 70, 5–17.

31.

Rong

, Hu

, Wu

, Cao

, and Chen

(2012). Interleukin-23 as a potential therapeutic target for rheumatoid arthritis. Mol Cell Biochem, 361, 243–248.

32.

Savvatis

, Pappritz

, Becher

P.M.

, Lindner

, Zietsch

, Volk

H.D.

, et al. (2014). Interleukin-23 deficiency leads to impaired wound healing and adverse prognosis after myocardial infarction. Circ Hear Fail, 7, 161–171.

33.

Tedgui

, and Mallat

(2006). Cytokines in atherosclerosis: pathogenic and regulatory pathways. Physiol Rev, 86, 515–581.

34.

Tindall

E.A.

, and Hayes

V.M.

(2010). Comprehensive sequence analysis of the human IL23A gene defines new variation content and high rate of evolutionary conservation. DNA Res, 17, 117–122.

35.

Vignali

D.A.A.

, and Kuchroo

V.K.

(2012). IL-12 family cytokines: immunological playmakers. Nat Immunol, 13, 722–728.

36.

Xia

L.P.

, Li

B.F.

, Shen

, and Lu

(2015). Interleukin-27 and interleukin-23 in patients with systemic lupus erythematosus: possible role in lupus nephritis. Scand. J Rheumatol, 44, 200–205.

37.

Yen

, Cheung

, Scheerens

, Poulet

, McClanahan

, McKenzie

, et al. (2006). IL-23 is essential for T cell-mediated colitis and promotes inflammation via IL-17 and IL-6. J Clin Invest, 116, 1310–1316.

38.

, Lee

D.-S.

, Jeon

M.-S.

, Kwon

S.W.

, and Song

S.U.

(2012). Gene expression profile reveals that STAT2 is involved in the immunosuppressive function of human bone marrow-derived mesenchymal stem cells. Gene, 497, 131–139.