rs2569190A>G in CD14 is Independently Associated with Hypercholesterolemia: A Brief Report

Journal of Cardiovascular Development and Disease Brief Report rs2569190A>G in CD14 is Independently Associated with Hypercholesterolemia: A Brief Report Ali Salami 1 , Christy Costanian 2,3 and Said El Shamieh 4, * 1 2 3 4 * Rammal Hassan Rammal Research Laboratory, Physio-toxicity (PhyTox) Research Group, Lebanese University, Faculty of Sciences (V), Nabatieh 1700, Lebanon; Faculty of Medicine, University of Ottawa, Ottawa 61350, ON K1G 5Z3, Canada; School of Medicine, Lebanese American University, Beirut 1102 2801, Lebanon Department of Medical Laboratory Technology, Faculty of Health Sciences, Beirut Arab University, Beirut 115020, Lebanon Correspondence: ; Tel.: +961-1-300110 (ext. 2721) Academic Editor: Maurice Van den Hoff Received: 5 September 2019; Accepted: 28 October 2019; Published: 30 October 2019



 Abstract: Many studies have assessed the implication of cluster of differentiation 14 (CD14) molecules and its single nucleotide polymorphism rs2569190A>G with different complex diseases, such as diabetes and cardiovascular diseases (CVDs). In this study, we investigated the association of rs2569190A>G in CD14 with cardiovascular disease risk factors (hypercholesterolemia and hypertension) in 460 individuals from the general Lebanese population (Middle Eastern multiethnic population). Using a multiple logistic regression model adjusted for six covariates (under additive and recessive assumptions), we found that the G allele of rs2569190 in CD14 was associated with increased levels of total cholesterol (OR = 3.10, p = 0.009), low-density lipoprotein cholesterol (OR = 3.87, p = 0.003), and decreased levels of high-density lipoprotein cholesterol (OR = 0.38, p = 0.001). In contrast, no significant relationship was found with hypertension. Thus, we concluded that rs2569190G in CD14 is associated with a higher risk of developing hypercholesterolemia. Keywords: hypercholesterolemia; cluster of differentiation 14; rs2569190A>G; single nucleotide polymorphisms; association analysis 1. Introduction According to the World Health Organization, cardiovascular diseases (CVDs) represent the most significant cause of death in humans globally [1]. Around 18 million people died because of CVDs in 2016, representing 31% of all deaths worldwide, most of which occur in low- and middle-income countries [1]. In addition to modifiable risk factors such as high cholesterol and triglyceride levels, diabetes, and high blood pressure (BP) levels [2], several studies have revealed an important impact of the innate immune system on the development or the progression of many CVDs [3]. The innate immune system present in multicellular living organisms gives an immediate defense capability against foreign bodies and pathogenic organisms such as viruses, bacteria, and fungi at first exposure [4]. This system depends on the recognition of pathogens by several families of extracellular receptors, such as the cluster of differentiation 14 (CD14), which is responsible for triggering innate immune responses and was first identified as marker of monocytes, before being defined as a coreceptor of toll-like receptors [5]. Many studies have assessed the role of CD14, especially at the molecular level, and established a link between the gene product and its single nucleotide polymorphisms (SNPs) with different complex diseases such as diabetes [6] and CVDs [7]. For example, the SNP rs2569190 in the promoter region of CD14 has been found to be implicated in coronary artery disease through changing protein levels [8,9]. J. Cardiovasc. Dev. Dis. 2019, 6, 37; doi:10.3390/jcdd6040037 www.mdpi.com/journal/jcdd J. Cardiovasc. Dev. Dis. 2019, 6, 37 2 of 6 Furthermore, the A allele has been reported to be functional and enhances CD14 expression, and thus the host sensitivity, for exogenous or endogenous lipopolysaccharides [10]. More importantly, soluble CD14 levels have shown strong, independent correlations with traditional cardio-metabolic risk factors and with subclinical measures of vascular disease (carotid wall thickness, ankle-brachial index, and body mass index) [7]. Based on all of the above, we hypothesize that rs2569190 in CD14 could be associated with CVD risk factors such as hypercholesterolemia and hypertension (HTN). Therefore, the goal of our study was to investigate the association of rs2569190 in CD14 with CVD risk factors in individuals from the general Lebanese population. 2. Materials and Methods 2.1. Study Population The institutional review board of the Lebanese University approved the recruitment procedure and the genetic protocols (2182/28, on 16 December 2015). This cross-sectional study involved 460 unrelated Lebanese participants who were apparently healthy (free of chronic diseases; cardiovascular or cancer) individuals. 2.2. Clinical and Biological Data Collection All measurements, including demographic, clinical, and biochemical measurements, were assessed as described previously [11]. Nuclear DNA was extracted from whole-blood samples according to the manufacturer’s recommendations (QIAamp DNA blood mini kit, Qiagen, Hilden, Germany). Very briefly, 4 µg of total DNA from 200 µL of whole human blood was extracted through lysis and continuous spinning. A KASP genotyping assay (LGC group, Berlin, Germany) was used to genotype rs2569190A>G in CD14. Hypercholesterolemia was defined as an elevation of total cholesterol (Tchol) and/or low-density cholesterol (LDL-C) levels. Tchol and LDL-C were considered high if their values were ≥150 and ≥100 mg/dL, respectively. High-density cholesterol (HDL-C) levels were considered low if their values were ≤50 and ≤40 mg/dL in females and males, respectively. HTN was defined as systolic blood pressure ≥130 mmHg or diastolic ≥85 mmHg. 2.3. Statistical Analyses SPSS statistical software version 24.0 [12] was used to perform all our statistical analyses except the power analysis. GPower 3.1.9.4 software [13] was used for the power analysis. Continuous variables were presented as mean value ± standard deviation, and categorical ones were shown as numbers and percentages. A chi-squared goodness-of-fit test was performed to determine if the genotypes of rs2569190A>G in CD14 were in Hardy–Weinberg equilibrium (HWE). To study the association between rs2569190A>G in CD14 and hypercholesterolemia and HTN, a multivariate logistic regression model was used while correcting for different confounding factors (age, gender, body mass index, marital status, smoking, and physical activity). This analysis was performed under the assumption of additive (AA vs. GA vs. GG) and recessive models (AA and GA vs. GG). The sample size needed to reach a statistical power of at least 0.90 in a two-sided test with α = 0.05 and an effect size of 0.2 was 409 individuals. 3. Results 3.1. Characteristics of the Studied Participants The demographic characteristics of the study participants are shown in Table 1. The group of participants comprised 292 (...truncated)