title: Association of frequent genetic variants in platelet activation pathway genes with large-vessel ischemic stroke in Polish population.
abstract: Platelets are critically involved in the development of cerebral ischemia. Our study aimed to establish an association between frequent (minor allele frequency (MAF) > 5%) genetic polymorphisms in 84 candidate genetic loci previously linked to platelet reactivity by the use of next-generation sequencing of exons from pooled DNA samples in Polish patients with a history of large-vessel ischemic stroke. Genetic analysis was performed on blood samples obtained from 500 patients (diagnosed with acute non-cardioembolic ischemic stroke with coexisting large-artery atherosclerosis) and age/sex/history of smoking matching 500 controls of Polish origin with high risk of cardiovascular disease. Sequencing of 10 pools (five for each ischemic and control groups) was performed on the Ilumina HiSeq2500 sequencer which generated an average of 36.1 (22.7-45.9 range) million pair-end 101 bp reads and 5.3 (3-7 range) Gbp per pooled sample consisting of 100 subjects. In total, we observed 789 frequent polymorphisms in the sequenced 84 genes (703 of single-nucleotide polymorphism (SNP) type and 86 indels). When the MAF between control and stroke groups was compared, only two intronic polymorphisms (1 SNP and 1 indel) in RGS7 (rs127445 36) and ANKS1B (rs398098426) genes, respectively, show statistically significant differences, which persisted after individual genotyping of the variants and adjustment for potential confounding factors. From the remaining variants, 35 polymorphisms displayed various degrees of nominal significance (from 0.6.3 × 10-5to 5 × 10-2) and 754 polymorphisms did not show any statistical significance when comparison was evaluated for differences in MAF between the study groups. In conclusion, the results of the study demonstrate statistically significant differences in two frequent intronic genetic variants (in RGS7 and ANKS1B) that could be associated with the platelet function between ischemic stroke patients with coexisting large-vessel atherosclerosis and control patients having high vascular risk.
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 University, College of Medicine , Hershey , PA , USA.
authors.forename: Marek
authors.initials: M
authors.lastname: Postula
authors.affiliation: b Perioperative Genomics Laboratory , Penn State University, 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: Lukasz
authors.initials: L
authors.lastname: Milanowski
authors.affiliation: a Department of Experimental and Clinical Pharmacology , Medical University of Warsaw, Center for Preclinical Research and Technology CEPT , Warsaw , Poland.
authors.forename: Justyna
authors.initials: J
authors.lastname: Pordzik
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: c 2nd Department of Neurology , Institute of Psychiatry and Neurology , Warsaw , Poland.
authors.forename: Michal
authors.initials: M
authors.lastname: Karlinski
authors.affiliation: a Department of Experimental and Clinical Pharmacology , Medical University of Warsaw, Center for Preclinical Research and Technology CEPT , Warsaw , Poland.
authors.forename: Pawel
authors.initials: P
authors.lastname: Wylezol
authors.affiliation: a Department of Experimental and Clinical Pharmacology , Medical University of Warsaw, Center for Preclinical Research and Technology CEPT , Warsaw , Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Solarska
authors.affiliation: a Department of Experimental and Clinical Pharmacology , Medical University of Warsaw, Center for Preclinical Research and Technology CEPT , Warsaw , Poland.|c 2nd Department of Neurology , Institute of Psychiatry and Neurology , Warsaw , Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Czlonkowka
authors.affiliation: c 2nd Department of Neurology , Institute of Psychiatry and Neurology , Warsaw , Poland.
authors.forename: Iwona
authors.initials: I
authors.lastname: Kurkowska-Jastrzebka
authors.affiliation: b Perioperative Genomics Laboratory , Penn State University, College of Medicine , Hershey , PA , USA.
authors.forename: Shigekazu
authors.initials: S
authors.lastname: Sugino
authors.affiliation: d Genome Sciences Facility , Penn State University, College of Medicine , Hershey , PA , USA.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Imamura
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; D000369:Aged, 80 and over; D000483:Alleles; D002545:Brain Ischemia / Q000150:complications* / Q000235:genetics*; D016022:Case-Control Studies; D005260:Female; D005787:Gene Frequency; D056726:Genetic Association Studies; D020022:Genetic Predisposition to Disease; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D016017:Odds Ratio; D015539:Platelet Activation / Q000235:genetics*; D011044:Poland / Q000453:epidemiology; D020641:Polymorphism, Single Nucleotide; D012307:Risk Factors; D020521:Stroke / Q000453:epidemiology / Q000209:etiology*
keywords: Genetics; ischemic stroke; next-generation sequencing; platelets activation