title: Next-generation re-sequencing of genes involved in increased platelet reactivity in diabetic patients on acetylsalicylic acid.
abstract: The objective of this study was to investigate whether rare missense genetic variants in several genes related to platelet functions and acetylsalicylic acid (ASA) response are associated with the platelet reactivity in patients with diabetes type 2 (T2D) on ASA therapy. Fifty eight exons and corresponding introns of eight selected genes, including PTGS1, PTGS2, TXBAS1, PTGIS, ADRA2A, ADRA2B, TXBA2R, and P2RY1 were re-sequenced in 230 DNA samples from T2D patients by using a pooled PCR amplification and next-generation sequencing by Illumina HiSeq2000. The observed non-synonymous variants were confirmed by individual genotyping of 384 DNA samples comprising of the individuals from the original discovery pools and additional verification cohort of 154 ASA-treated T2DM patients. The association between investigated phenotypes (ASA induced changes in platelets reactivity by PFA-100, VerifyNow and serum thromboxane B2 level [sTxB2]), and accumulation of rare missense variants (genetic burden) in investigated genes was tested using statistical collapsing tests. We identified a total of 35 exonic variants, including 3 common missense variants, 15 rare missense variants, and 17 synonymous variants in 8 investigated genes. The rare missense variants exhibited statistically significant difference in the accumulation pattern between a group of patients with increased and normal platelet reactivity based on PFA-100 assay. Our study suggests that genetic burden of the rare functional variants in eight genes may contribute to differences in the platelet reactivity measured with the PFA-100 assay in the T2DM patients treated with ASA.
authors.affiliation: a Department of Experimental and Clinical Pharmacology , Medical University of Warsaw, Center for Preclinical Research and Technology CEPT , Warsaw , Poland.|b Perioperative Genomics Laboratory , Penn State College of Medicine , Hershey , PA , USA.
authors.forename: Marek
authors.initials: M
authors.lastname: Postula
authors.affiliation: b Perioperative Genomics Laboratory , Penn State College of Medicine , Hershey , PA , USA.
authors.forename: Piotr K
authors.initials: PK
authors.lastname: Janicki
authors.affiliation: a Department of Experimental and Clinical Pharmacology , Medical University of Warsaw, Center for Preclinical Research and Technology CEPT , Warsaw , Poland.
authors.forename: Ceren
authors.initials: C
authors.lastname: Eyileten
authors.affiliation: a Department of Experimental and Clinical Pharmacology , Medical University of Warsaw, Center for Preclinical Research and Technology CEPT , Warsaw , Poland.|c Department of Cardiology and Hypertension , Central Clinical Hospital, The Ministry of the Interior , Warsaw , Poland.
authors.forename: Marek
authors.initials: M
authors.lastname: Rosiak
authors.affiliation: d Department of Cardiology , Medical University of Warsaw , Warsaw , Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Kaplon-Cieslicka
authors.affiliation: b Perioperative Genomics Laboratory , Penn State College of Medicine , Hershey , PA , USA.
authors.forename: Shigekazu
authors.initials: S
authors.lastname: Sugino
authors.affiliation: e Department of Cardiac Surgery , Medical University of Warsaw , Warsaw , Poland.
authors.forename: Radosław
authors.initials: R
authors.lastname: Wilimski
authors.affiliation: c Department of Cardiology and Hypertension , Central Clinical Hospital, The Ministry of the Interior , Warsaw , Poland.|f Department of Applied Physiology , Mossakowski Medical Research Centre, Polish Academy of Sciences , Warsaw , Poland.
authors.forename: Dariusz A
authors.initials: DA
authors.lastname: Kosior
authors.affiliation: d Department of Cardiology , Medical University of Warsaw , Warsaw , Poland.
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Opolski
authors.affiliation: d Department of Cardiology , Medical University of Warsaw , Warsaw , Poland.
authors.forename: Krzysztof J
authors.initials: KJ
authors.lastname: Filipiak
authors.affiliation: a Department of Experimental and Clinical Pharmacology , Medical University of Warsaw, Center for Preclinical Research and Technology CEPT , Warsaw , Poland.
authors.forename: Dagmara
authors.initials: D
authors.lastname: Mirowska-Guzel
mesh_terms: D000368:Aged; D000483:Alleles; D001241:Aspirin / Q000494:pharmacology*; D015415:Biomarkers; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism*; D003924:Diabetes Mellitus, Type 2 / Q000097:blood* / Q000188:drug therapy / Q000235:genetics*; D005091:Exons; D005260:Female; D005787:Gene Frequency; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense; D015539:Platelet Activation / Q000187:drug effects* / Q000235:genetics*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D020641:Polymorphism, Single Nucleotide; D012307:Risk Factors
chemical_list: D015415:Biomarkers; D010975:Platelet Aggregation Inhibitors; D001241:Aspirin
keywords: Acetylsalicylic acid; diabetes mellitus; genetic polymorphism; next-generation sequencing; platelet reactivity