Lack of effect of common single nucleotide polymorphisms in leukotriene pathway genes on platelet reactivity in patients with diabetes

Abstract

Abstract. The aim of the present study was to investigate the effect of genetic polymorphisms in candidate genes within the leukotriene (LT) pathway on platelet reactivity and the concentration of selected LTs in diabetic patients treated with acetylsalicylic acid (ASA). The study cohort consisted of 287 Caucasians with type 2 diabetes who had received treatment with ASA tablets (75 mg/day) for at least three months. Platelet reactivity analyses were performed using VerifyNow aspirin and PFA-100 assays. The measured LTs included leukotriene B 4 (LTB 4 ) and leukotriene E 4 (LTE 4 ). Genotyping for the selected 25 single nucleotide polymorphisms (SNPs) within six genes of the LT pathway was performed using a Sequenom iPLEX platform. No statistically significant association was observed between the investigated SNP genotypes, platelet reactivity and measured LTs in the patient cohort. The results of our study suggest that certain polymorphisms of the LT pathway are not associated with altered platelet reactivity and the measured LTs in diabetic patients treated with ASA.

Targeted deep resequencing of ALOX5 and ALOX5AP in patients with diabetes and association of rare variants with leukotriene pathways

“…However, the majority of previous studies have investigated the common polymorphic variants in the ALOX5 and ALOX5AP genes, as neither gene exhibits any significant common variants in their coding sequences. The results of a previous study by the present authors was unable to confirm whether the selected 14 common intronic variants in the ALOX5AP gene within the LT metabolism pathway contribute to platelet reactivity in a diabetic population treated with ASA (20).…”

“…(Table I) (19). The full characterization of the study population, including the inclusion and exclusion criteria were published previously (20,21). DNA samples were collected from 303 Caucasian subjects with T2DM that had been administered ASA tablets (75 mg per day) for ≥3 months at the time of enrollment for primary or secondary prevention of myocardial infarction (MI).…”

“…Genomic DNA (gDNA) was extracted from venous ethylenediaminetetraacetic acid (EDTA) antecubital vein-whole blood sample using a FujiFilm Life Sciences membrane ultrafiltration method (AutoGen, Holliston, MA, USA) (19)(20)(21), according to the manufacturer's recommendations. The selected number of gDNA samples were pooled in an equimolar manner and verified according to the previously described method from our laboratory (19).…”

“…501060 and 520111, respectively; Cayman Chemical Co.) following extraction and purification using solid phrase extraction (C18) columns (Waters Corporation, Milford, MA, USA). Data were normalized against urinary creatinine concentrations (20).…”

“…Genotyping for selected markers in individual DNA samples was performed at the Boston Children's Hospital (Boston, MA, USA) using a custom Sequenom iPLEX assay in conjunction with a MassARRAY platform (Sequenom Inc., La Jolla, CA, USA). Panels of SNP markers were designed using Sequenom Assay Design software, version 3.2 (Sequenom, Inc.) (20,21,32).…”