A common polymorphism in CD40 Kozak sequence (-1C/T) is associated with acute coronary syndrome

doi:10.1016/j.biopha.2009.09.021

Biomedicine & Pharmacotherapy

Volume 64, Issue 3, March 2010, Pages 191-194

https://doi.org/10.1016/j.biopha.2009.09.021 Get rights and content

Abstract

Objective

Evidence suggests the CD40-CD40L pathway as a key process in the development, progression, and outcome of acute coronary syndrome (ACS). We hypothesized that the -1C/T polymorphism of the CD40 gene would be associated with ACS and influence the CD40 expression.

Methods

The genotype distribution and allele frequency of CD40-1C/T polymorphism in 248 ACS patients and 206 controls were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Monocytes from 90 healthy volunteers were incubated with IFN-γ. CD40 expression was detected by flow cytometry.

Result

Patients with ACS showed a significant increase of CD40 expression compared with controls (P < 0.001). The frequency of the CC genotype in the ACS group was significantly higher than that of the controls (P < 0.001). Frequency of the C allele was higher among ACS patients compared with controls (P < 0.001). Case control association analysis of the CD40 -1C/T SNP showed an association between the C allele and ACS (OR = 1.991, 95%CI: 1.526 ∼ 2.596, P < 0.001). -1C/C carriers presented significantly higher CD40 expression levels than -1C/T and -1T/T subjects, both in ACS group and controls (P < 0.001). When stimulated by IFN-γ, CD40 expression levels on monocytes in individuals with CC, CT and TT genotypes were increased by 9.16, 3.83 and 1.53 fold, respectively, compared with the levels absent with IFN-γ.

Conclusions

These results suggest that the -1C allele of the CD40 (-1C/T) gene polymorphism is a genetic factor that may determine an individual's susceptibility by ACS in Chinese. The CD40 -1C/T polymorphism is a novel regulator of CD40 expression.

Introduction

Acute coronary syndrome (ACS) is currently considered the result of atherosclerotic plaques’ disruption. Several studies clearly demonstrate that CD40 is a major trigger, eliciting a proinflammatory reaction in the vasculature, and plays an important role in the formation of atherosclerotic lesion [1]. This can be evidenced by the finding that inhibition of CD40/CD40L interaction reduces atherosclerosis in mice [2], [3]. Recent studies strongly suggest that in the “shoulder” regions of lipid-rich plaques, which are known to contain dense inflammatory infiltrates, CD40 are positively expressed. Patients with ACS showed a significant increase of CD40 and CD40L coexpression on platelets compared with control and SA group [4]. These dates prove that the CD40-CD40L system plays a crucial role in the pathomechanism of ACS. It has been proved that a C/T polymorphism in the Kozak sequence of the CD40 gene enhances the efficiency of CD40 gene translation [5]. However, the relationship between the polymorphism and ACS and whether the C/T single nuclear polymorphism could influence the CD40 expression on B cells remain little reported. Therefore, we established a case-control study to analyse the association of ACS and C/T polymorphism in the CD40 gene, as well as the CD40 expression on B cells and monocytes.

Section snippets

Patients with ACS

Two hundred and forty eight patients with ACS were collected, which includes Q-wave myocardial infarction, non Q-wave myocardial infarction or unstable angina. Q-wave myocardial infarction was defined by the presence of (1) chest pain typical for myocardial ischemia lasting > 30 min, (2) elevation of total creatine kinase to at least twice the upper limit of normal with an elevated creatine kinase-MB fraction, and (3) development of diagnostic Q waves in ≥ 2 electrically contiguous leads.

Clinical and laboratory characteristics in the patients and controls

The characteristics of the patient and control groups are shown in Table 1. The ACS patients are found to be more likely to have hypertension and diabetes. Lipid profiles revealed that the ACS patient group have a significantly lower mean level of HDL- cholesterol and apolipoprotein AI (P < 0.01). SBP, DBP and Lp(a) are significantly higher in the ACS patients than that in the controls. Sex, age, BMI, FBG, total cholesterol, triglycerides, LDL- cholesterol, and apolipoprotein B levels are not

Discussion

CD40, a 50-KD integral membrane protein of the tumor necrosis factor receptor family, and CD40L are found positively expressed in the “shoulder” of atheromatous plaque by immunohistochemistry. Recently, increasing evidence shows that the CD40-CD40L system plays a critical role in atherosclerotic disease progression and plaque destabilization [8].

The CD40-CD40L system has been accepted to be involved in the formation, development and outcome of ACS. Engagement of the endothelial CD40 receptor by

Conflicts of interest

None.

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The CD40 receptor and its ligand CD154 are widely expressed in various immune-competent cells. Interaction of CD154 with CD40 is essential for B-cell growth, differentiation, and immunoglobulin class switching. Many other immune-competent cells involved in innate and adaptive immunity communicate through this co-stimulatory ligand-receptor dyad. CD40-CD154 interaction is involved in the pathogenesis of numerous inflammatory and autoimmune diseases. While CD40 and CD154 are membrane-bound proteins, their soluble counterparts are generated by proteolytic cleavage or alternative splicing.
This review summarises current knowledge about the impact of single nucleotide polymorphisms in the human CD40 gene and compensatory changes in the plasma level of the soluble CD40 receptor (sCD40) isoform in related pro-inflammatory diseases. It discusses regulation patterns of the disintegrin metalloprotease ADAM17 function leading to ectodomain shedding of transmembrane proteins, such as pro-inflammatory adhesion molecules or CD40. The role of sCD40 as a potential biomarker for chronic inflammatory diseases will also be discussed.
Lack of association between CD40 polymorphisms and acute rejection in German liver transplant recipients
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One study reported for the SNP rs1883832 a difference in allele frequency of 20% resulting in association for Graves’ disease intractability [26]. Further studies described for the selected SNPs different kind of associations based on allele differences near 19% [19,27,28]. Regarding those data, our study appears sufficiently powered to replicate the above reported associations.
CD40 and its ligand, CD154, are major costimulatory molecules whose interactions are important in alloreactive transplant rejection. The aim of this study was to examine the association of CD40 polymorphisms with the susceptibility to acute rejection episodes in liver transplantation. In total, 112 liver transplant recipients with biopsy proven acute rejections (BPAR), 97 without BPAR (WBPAR), and 112 healthy control individuals were enrolled in the study. Two single nucleotide polymorphisms (SNPs) of CD40 gene (rs1883832 and rs4810485) were genotyped by polymerase chain reaction-allele specific restriction enzyme analysis (PCR-ASRA). Both SNPs has been tested for a recessive and a dominant model. No significant differences were found in the genotype and allele frequencies of the SNPs rs1883832 and rs4810485 between BPAR liver recipients and WBPAR recipients. Our results do not suggest an important role of tested CD40 SNPs in the susceptibility to acute liver transplant rejection in a Caucasian population.
Evidence for involvement of the CD40/CD40L system in post-stroke epilepsy
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Post-stroke epilepsy (PSE) has a negative effect on stroke prognosis and quality of life. The CD40/CD40L system is reported to be involved in the progression of multiple disease states. We investigated the association between functional polymorphism of CD40 and PSE susceptibility, and we also explored the role of the CD40/CD40L system in PSE. A case-control study was performed in 410 ischemic stroke (IS) patients and in 389 PSE patients. Genotyping was performed by polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP). The CD40 mRNA and protein levels were determined by real-time PCR and western blotting, respectively. The plasma sCD40L level was detected using an ELISA kit. The frequency of the T allele in PSE patients was significantly higher than in IS patients (P < 0.05). The plasma sCD40L level was significantly higher in the PSE patients than in the healthy controls and IS patients (P < 0.01, P < 0.05, respectively). The peripheral blood mononuclear cells (PBMCs) from PSE patients showed significantly higher CD40 mRNA and protein expression than the healthy controls and IS patients (P < 0.01, P < 0.05, respectively). In the PSE patients, the T-allele carriers showed increased plasma sCD40L levels and increased CD40 mRNA expression. Our study suggested that the T allele of the CD40 −1C/T polymorphism may be associated with PSE susceptibility. The CD40/CD40L system is involved in the process of PSE.
Single nucleotide polymorphism of CD40 in the 5'-untranslated region is associated with ischemic stroke
2013, Gene
Citation Excerpt :
There are about 45 SNPs for the CD40 gene in the Chinese Han popula-tion (http://genome.ucsc.edu/cgi-bin/hgTracks). CD40 SNPs associated with infection- and autoimmunity-associated diseases were widely studied, e.g. acute coronary syndrome, Graves' disease (Burdon et al., 2006; Kurylowicz et al., 2005; Tian et al., 2010). However, only one known SNP affected CD40 protein levels, which was located at the -1 position in the 5′-untranslated region (5′-UTR) nearby the start codon, and coincided with the Kozak consensus sequence(CD40 -1C/T; refSNP ID: rs1883832) (Jacobson et al., 2005; Kozak, 1987; Tomer et al., 2002).
Ischemic stroke is influenced by both environmental and genetic factors. The CD40/CD40L system is related to proinflammatory and prothrombogenic responses, which are involved in the pathophysiology of ischemic stroke. The aim of this study was to evaluate association between the CD40 -1C/T single nucleotide polymorphism (SNP) and ischemic stroke in a Chinese population.
We conducted a case–control study including 286 ischemic stroke patients and 336 controls. CD40 -1C/T SNP was genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing methods, and evaluated its relevance to ischemic stroke susceptibility.
Significantly increased ischemic stroke risk was found to be associated with the T allele of CD40 -1C/T (OR = 1.273, 95% CI = 1.016–1.594). The frequencies of CT and TT/CT genotypes of CD40 -1C/T in ischemic stroke patients were significantly higher than those of controls, respectively (for CT: OR = 2.350, 95% CI = 1.601–3.449; for TT/CT: OR = 2.148, 95% CI = 1.479–3.119). And, similar results were obtained after adjusting non-matched variables. We found that the frequency of carried T genotypes (TT and TT/CT) was significantly increased in patients with history of stroke compared with patients without (for TT: OR = 6.538, 95%CI = 1.655–25.833; for TT/CT: OR = 3.469, 95%CI = 1.031–11.670), respectively.
The findings suggested that the CD40 -1C/T polymorphism might contribute to the susceptibility to ischemic stroke in the Chinese population, and might be associated with history of previous stroke.
Association of CD40 - 1C/T polymorphism with cerebral infarction susceptibility and its effect on sCD40L in Chinese population
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This study aims to determine whether functional polymorphism of CD40 is associated with the cerebral infarction (CI) susceptibility, and to investigate the effect of CD40 gene polymorphism on CD40 mRNA expression in PBMCs and plasma sCD40L concentration. A case–control study was performed in 402 CI patients and 693 controls. Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The expressions of CD40 mRNA and plasma sCD40L concentration were determined. The distribution of TT genotype and the frequency of T allele in CI patients were significantly higher than those in the controls (P < 0.05). The frequency of T allele was also significantly higher in the male subjects and the elder subjects (P < 0.05) when stratified analysis was carried out. The PBMCs from CI patients showed significantly higher CD40 mRNA expression than controls (P < 0.01), the CD40 mRNA expression from TT genotype was higher than other genotypes (P < 0.05). TT genotype subjects also showed the highest plasma sCD40L concentration in the male CI patients (P < 0.01). CD40 − 1C/T polymorphism may interfere CI susceptibility, and the T allele may be associated with increased risk of CI. The CD40 − 1C/T polymorphism is also a regulator of CD40 expression and plasma sCD40L concentration.
From Atherosclerosis to Acute Coronary Syndromes: The Role of Soluble CD40 Ligand
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Citation Excerpt :
Recently, two studies in Chinese populations of patients with ACS revealed that the −1C/T polymorphism of the CD40 gene was associated with ACS. The C/T in the 5′ untranslated region of CD40 located at the −1 position (rs1883832) has been associated with CD40 protein expression in previous studies (Tian et al. 2010; Jacobson et al. 2005). It has been reported that patients with ACS exhibited a significant up-regulation of CD40L expression compared with controls.
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Original articleA common polymorphism in CD40 Kozak sequence (-1C/T) is associated with acute coronary syndrome

Abstract

Objective

Methods

Result

Conclusions

Introduction

Section snippets

Patients with ACS

Clinical and laboratory characteristics in the patients and controls

Discussion

Conflicts of interest

Chin Med J

Am J Pathol

Gene

CD40 and vascular inflammation

Can J Cardiol

Reduction of atherosclerosis in mice by inhibition of CD40 signalling

Nature

Inhibition of CD40 signaling limits evolution of established atherosclerosis in mice

Proc Natl Acad Sci U S A

Relation between upregulation of CD40 system and complex stenosis morphology in patients with acute coronary syndrome

Acta Pharmacol Sin

A Graves’ disease-associated Kozak sequence single-nucleotide polymorphism enhances the efficiency of CD40 gene translation: a case for translational pathophysiology

Endocrinology

Original article
A common polymorphism in CD40 Kozak sequence (-1C/T) is associated with acute coronary syndrome