Yu Lei, Hou Ping, Lv Ji-Cheng, Liu Guo-Ping, Zhang Hong
Renal Division, Inner Mongolia People's Hospital, Hohhot, People's Republic of China.
Hum Mutat. 2015 Jan;36(1):79-86. doi: 10.1002/humu.22714. Epub 2014 Nov 28.
Familial renal glucosuria (FRG) is characterized by persistent glucosuria despite normal serum glucose and the absence of overt tubular dysfunction. Variants in solute carrier family 5 (sodium-glucose cotransporter), member 2 (SLC5A2) have been reported in FRG patients. However, the functional and expression-related consequences of such variants have been scarcely investigated. In the current study, we studied five FRG families and identified six missense mutations, including four novel variants (c.1051T>C/.(C351R), c.1400T>C/p.(V467A), c.1420G>C/p.(A474P), c.1691G>A/p.(R564Q); RNA not analyzed) and two variants that had been previously reported (c.294C>A/p.(F98L), c.736C>T/p.(P246S); RNA not analyzed). The probands were either heterozygous or compound heterozygous for SLC5A2 variants and had glucosuria of 5.9%-19.6 g/day. Human 293 cells were transfected with plasmid constructs to study the expression and function of SLC5A2 variants in vitro. Western blotting revealed that the expression levels of SLC5A2-351R-GFP, SLC5A2-467A-GFP, SLC5A2-474P-GFP, and SLC5A2-564Q-GFP were significantly decreased compared with wild-type SLC5A2-GFP (37%-55%). Confocal microscopy revealed that three variants (c.1400T>C, c.1420G>C, c.1691G>A) resulted in a loss of the punctate membrane pattern typical of wild-type SLC5A2. All variants had a significantly lower transport capacity in than the wild-type control. The current study provides a starting point to further investigate the molecular mechanism of SLC5A2 in FRG families and provides functional clues for antidiabetes drugs.
家族性肾性糖尿(FRG)的特征是尽管血清葡萄糖正常且无明显肾小管功能障碍,但仍持续出现糖尿。溶质载体家族5(钠-葡萄糖共转运蛋白)成员2(SLC5A2)的变异已在FRG患者中被报道。然而,此类变异的功能及与表达相关的后果鲜有研究。在本研究中,我们研究了五个FRG家族,鉴定出六个错义突变,包括四个新变异(c.1051T>C/p.(C351R)、c.1400T>C/p.(V467A)、c.1420G>C/p.(A474P)、c.1691G>A/p.(R564Q);未分析RNA)和两个先前已报道的变异(c.294C>A/p.(F98L)、c.736C>T/p.(P246S);未分析RNA)。先证者为SLC5A2变异的杂合子或复合杂合子,糖尿量为5.9%-19.6克/天。用质粒构建体转染人293细胞以在体外研究SLC5A2变异的表达和功能。蛋白质印迹显示,与野生型SLC5A2-GFP相比,SLC5A2-351R-GFP、SLC5A2-467A-GFP、SLC5A2-474P-GFP和SLC5A2-564Q-GFP的表达水平显著降低(37%-55%)。共聚焦显微镜检查显示,三个变异(c.1400T>C、c.1420G>C、c.1691G>A)导致野生型SLC5A2典型的点状膜模式消失。所有变异的转运能力均显著低于野生型对照。本研究为进一步研究FRG家族中SLC5A2的分子机制提供了一个起点,并为抗糖尿病药物提供了功能线索。