Genetic variations in urate transporters play a significant role in determining human urate levels and have been implicated in developing hyperuricemia or gout. Polymorphism in the key urate transporters, such as ABCG2, URAT1, or GLUT9 was well-documented in the literature. Therefore in this study, our objective was to determine the frequency and effect of rare nonsynonymous allelic variants of SLC22A11, SLC22A13, and SLC17A1 on urate transport. In a cohort of 150 Czech patients with primary hyperuricemia and gout, we examined all coding regions and exon–intron boundaries of SLC22A11, SLC22A13, and SLC17A1 using PCR amplification and Sanger sequencing. For comparison, we used a control group consisting of 115 normouricemic subjects. To examine the effects of the rare allelic nonsynonymous variants on the expression, intracellular processing, and urate transporter protein function, we performed a functional characterization using the HEK293A cell line, immunoblotting, fluorescent microscopy, and site directed mutagenesis for preparing variants in vitro. Variants p.V202M (rs201209258), p.R343L (rs75933978), and p.P519L (rs144573306) were identified in the SLC22A11 gene (OAT4 transporter); variants p.R16H (rs72542450), and p.R102H (rs113229654) in the SLC22A13 gene (OAT10 transporter); and the p.W75C variant in the SLC17A1 gene (NPT1 transporter). All variants minimally affected protein levels and cytoplasmic/plasma membrane localization. The functional in vitro assay revealed that contrary to the native proteins, variants p.P519L in OAT4 (), p.R16H in OAT10 (), and p.W75C in the NPT1 transporter () significantly limited urate transport activity. Our findings contribute to a better understanding of (1) the risk of urate transporter-related hyperuricemia/gout and (2) uric acid handling in the kidneys.

中文翻译：

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检查尿酸盐转运蛋白 SLC22A11、SLC22A13 和 SLC17A1 中罕见等位基因变异与高尿酸血症和痛风的关联

尿酸转运蛋白的遗传变异在决定人类尿酸水平方面发挥着重要作用，并且与高尿酸血症或痛风的发生有关。关键尿酸转运蛋白（例如 ABCG2、URAT1 或 GLUT9）的多态性在文献中有详细记载。因此，在本研究中，我们的目标是确定SLC22A11、SLC22A13和SLC17A1的罕见非同义等位基因变异对尿酸盐转运的频率和影响。在 150 名捷克原发性高尿酸血症和痛风患者的队列中，我们使用 PCR 扩增和桑格测序检查了SLC22A11、SLC22A13和SLC17A1的所有编码区和外显子-内含子边界。为了进行比较，我们使用了由 115 名正常尿酸血症受试者组成的对照组。为了检查罕见等位基因非同义变体对表达、细胞内加工和尿酸转运蛋白功能的影响，我们使用 HEK293A 细胞系、免疫印迹、荧光显微镜和定点诱变进行功能表征，以在体外制备变体。在SLC22A11基因（OAT4 转运蛋白）中鉴定出变体 p.V202M (rs201209258)、p.R343L (rs75933978) 和 p.P519L (rs144573306) ；SLC22A13基因（OAT10 转运蛋白）中的变体 p.R16H (rs72542450) 和 p.R102H (rs113229654) ；以及SLC17A1基因（NPT1 转运蛋白）中的 p.W75C 变体。所有变体对蛋白质水平和细胞质/质膜定位的影响最小。功能性体外测定表明，与天然蛋白相反，OAT4 中的变体 p.P519L (）， OAT10 中的 p.R16H（），以及 NPT1 转运蛋白中的 p.W75C（）显着限制了尿酸盐转运活动。我们的研究结果有助于更好地了解（1）尿酸盐转运蛋白相关的高尿酸血症/痛风的风​​险和（2）肾脏中的尿酸处理。