Postula Marek, Janicki Piotr K, Eyileten Ceren, Rosiak Marek, Kaplon-Cieslicka Agnieszka, Sugino Shigekazu, Wilimski Radosław, Kosior Dariusz A, Opolski Grzegorz, Filipiak Krzysztof J, Mirowska-Guzel Dagmara
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.
Platelets. 2016 Jun;27(4):357-64. doi: 10.3109/09537104.2015.1109071. Epub 2015 Nov 24.
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.
本研究的目的是调查与血小板功能和乙酰水杨酸（ASA）反应相关的几个基因中的罕见错义遗传变异是否与接受ASA治疗的2型糖尿病（T2D）患者的血小板反应性相关。通过使用聚合酶链式反应（PCR）扩增和Illumina HiSeq2000二代测序技术，对230例T2D患者的DNA样本中8个选定基因（包括PTGS1、PTGS2、TXBAS1、PTGIS、ADRA2A、ADRA2B、TXBA2R和P2RY1）的58个外显子及相应内含子进行了重测序。通过对384个DNA样本进行个体基因分型，确认了观察到的非同义变异，这些样本包括来自原始发现库的个体以及另外154例接受ASA治疗的T2DM患者的验证队列。使用统计合并检验来测试所研究的表型（通过PFA-100、VerifyNow检测的ASA诱导的血小板反应性变化以及血清血栓素B2水平[sTxB2]）与所研究基因中罕见错义变异的累积（遗传负担）之间的关联。我们在8个研究基因中总共鉴定出35个外显子变异，包括3个常见错义变异、15个罕见错义变异和17个同义变异。基于PFA-100检测，在血小板反应性增加和正常的两组患者中，罕见错义变异在累积模式上表现出统计学显著差异。我们的研究表明，在接受ASA治疗的T2DM患者中，8个基因中罕见功能变异的遗传负担可能导致通过PFA-100检测测得的血小板反应性存在差异。
