Skarke Carsten, Schuss Patrick, Kirchhof Anja, Doehring Alexandra, Geisslinger Gerd, Lötsch Jörn
Johann Wolfgang Goethe-University, pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Theodor Stern Kai 7, 60590 Frankfurt, Germany.
Pharmacogenomics. 2007 Dec;8(12):1643-60. doi: 10.2217/14622416.8.12.1643.
Genetic variants in the prostaglandin-endoperoxide synthase 2 (PTGS2) gene, which codes for COX-2, have been identified to modulate the response to COX-2-inhibiting drugs and to be possible risk factors for the incidence or prognosis of cardiovascular or neoplastic diseases, Alzheimer's disease, multiple sclerosis, asthma or osteoarthritis. Clinical evidence thus suggests a clinical importance of COX-2 genetics reaching from disease risk or prognostics up to a personalized therapy with COX-2 inhibitors. The aim of this study was to develop rapid and reliable screening assays for PTGS2 mutations with reported clinical consequences.
SNPs (dbSNP-IDs rs689465, rs689466, rs3918304, rs20415, rs20417, rs5270, rs2745557, rs5277, rs2066826, rs4648276, rs5273, rs5275, rs4648298, rs689469) and a nucleotide-deletion variant (rs20431) were chosen according to reported functional associations. For this selection of variants spanning the whole PTGS2 gene range, Pyrosequencing assays were established in DNA from 350 healthy unrelated Caucasians.
In all 350 DNA samples, the 15 PTGS2 polymorphisms were identified correctly as verified by control samples obtained by conventional sequencing. In silico haplotype analysis based on ten SNPs of greater than 1% observed frequencies identified two haploblocks with a linkage disequilibrium of D' = 0.59. Approximately 50% of the reconstructed haplotypes consisted of non-mutated alleles.
The presently developed Pyrosequencing assays allow for quick and reliable detection of PTGS2 genotypes and may promote further research toward personalized approaches to pathophysiological conditions involving COX-2.
编码环氧化酶-2（COX-2）的前列腺素内过氧化物合酶2（PTGS2）基因中的遗传变异，已被证实可调节对COX-2抑制药物的反应，并且可能是心血管疾病、肿瘤疾病、阿尔茨海默病、多发性硬化症、哮喘或骨关节炎发病或预后的危险因素。因此，临床证据表明COX-2遗传学在疾病风险或预后以及COX-2抑制剂的个性化治疗方面具有临床重要性。本研究的目的是开发针对具有已报道临床后果的PTGS2突变的快速可靠筛选检测方法。
根据已报道的功能关联选择单核苷酸多态性（dbSNP ID：rs689465、rs689466、rs3918304、rs20415、rs20417、rs5270、rs2745557、rs5277、rs2066826、rs4648276、rs5273、rs5275、rs4648298、rs689469）和一个核苷酸缺失变异体（rs20431）。为了选择覆盖整个PTGS2基因范围的这些变异体，在来自350名健康无亲缘关系的白种人的DNA中建立了焦磷酸测序检测方法。
在所有350个DNA样本中，通过常规测序获得的对照样本验证，15个PTGS2多态性均被正确鉴定。基于观察频率大于1%的10个单核苷酸多态性进行的计算机单倍型分析确定了两个单倍型模块，其连锁不平衡D' = 0.59。大约50%的重建单倍型由未突变的等位基因组成。
目前开发的焦磷酸测序检测方法能够快速可靠地检测PTGS2基因型，并可能促进对涉及COX-2的病理生理状况的个性化方法的进一步研究。
