Elek Zsuzsanna, Németh Nóra, Nagy Géza, Németh Helga, Somogyi Anikó, Hosszufalusi Nóra, Sasvári-Székely Mária, Rónai Zsolt
Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary.
2nd Department of Internal Medicine, Semmelweis University, Budapest, Hungary.
PLoS One. 2015 Oct 1;10(10):e0139519. doi: 10.1371/journal.pone.0139519. eCollection 2015.
The absolute or relative lack of insulin is the key factor in the pathogenesis of diabetes mellitus. Although the connection between loss of function mutations of the WFS1 gene and DIDMOAD-syndrome including diabetes mellitus underpins the significance of wolframin in the pathogenesis, exact role of WFS1 polymorphic variants in the development of type 1 and type 2 diabetes has not been discovered yet. In this analysis, 787 patients with diabetes and 900 healthy people participated. Genotyping of the 7 WFS1 SNPs was carried out by TaqMan assays. Association study was performed by χ2-test in combination with correction for multiple testing. For functional analysis, the entire 3' UTR of the WFS1 gene was subcloned in a pMIR-Report plasmid and relative luciferase activities were determined. Linkage disequilibrium analysis showed a generally high LD within the investigated region, however the rs1046322 locus was not in LD with the other SNPs. The two miR-SNPs, rs1046322 and rs9457 showed significant association with T1DM and T2DM, respectively. Haplotype analysis also confirmed the association between the 3' UTR loci and both disease types. In vitro experiments showed that miR-185 reduces the amount of the resulting protein, and rs9457 miRSNP significantly influences the rate of reduction in a luciferase reporter assay. Genetic variants of the WFS1 gene might contribute to the genetic risk of T1DM and T2DM. Furthermore demonstrating the effect of rs9457 in binding of miR-185, we suggest that the optimal level of wolframin protein, potentially influenced by miR-regulation, is crucial in normal beta cell function.
胰岛素的绝对或相对缺乏是糖尿病发病机制中的关键因素。尽管WFS1基因功能丧失突变与包括糖尿病在内的DIDMOAD综合征之间的联系支持了沃尔弗拉姆蛋白在发病机制中的重要性，但WFS1多态性变体在1型和2型糖尿病发生发展中的确切作用尚未明确。在本分析中，787例糖尿病患者和900名健康人参与研究。通过TaqMan分析对7个WFS1单核苷酸多态性（SNP）进行基因分型。采用χ2检验结合多重检验校正进行关联研究。为进行功能分析，将WFS1基因的整个3'非翻译区亚克隆到pMIR-Report质粒中，并测定相对荧光素酶活性。连锁不平衡分析显示，在研究区域内总体连锁不平衡程度较高，但rs1046322位点与其他SNP不存在连锁不平衡。两个微小RNA-SNP，rs1046322和rs9457分别与1型糖尿病和2型糖尿病显著相关。单倍型分析也证实了3'非翻译区位点与两种疾病类型之间的关联。体外实验表明，miR-185可减少产生的蛋白量，并且rs9457微小RNA-SNP在荧光素酶报告基因分析中显著影响减少速率。WFS1基因的遗传变异可能导致1型糖尿病和2型糖尿病的遗传风险。此外，通过证明rs9457对miR-185结合的影响，我们认为，可能受微小RNA调节影响的沃尔弗拉姆蛋白的最佳水平对正常β细胞功能至关重要。
