SLC26A2 function in the kidney

SLC26A2 在肾脏中的功能

• 批准号: 441931051
• 负责人: Professorin Dr. Ute Scholl
• 金额: --
• 依托单位: Center of Functional Genomics
• 依托单位国家: 德国
• 项目类别: Research Units
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/441931051?language=en
• 关键词: SLC26A2; function; kidney

Sulfate is essential for normal cellular function. For example, it is required for the sulfation of proteoglycans, molecules that contribute to the structure of normal cartilage. Loss-of-function mutations in a cartilage sulfate transporter (SLC26A2) are the cause of a group of autosomal recessive chondrodysplasias. In these disorders, impaired sulfate uptake into chondrocytes leads to reduced proteoglycan sulfation. However, SLC26A2 is not only expressed in cartilage, but also in other tissues, such as the kidney. There, it specifically localizes to the apical (luminal) brush border membrane of proximal tubular cells. Sulfate is known to be freely filtered and extensively reabsorbed in the kidney, in particular the proximal tubule, but the exact function of SLC26A2 for renal sulfate transport has not been explored.We here propose to explore the renal function and regulation of SLC26A2. In preliminary studies, using the CRISPR/Cas9 system, we knocked out SLC26A2 in a cellular model of the proximal tubule, OK (opossum kidney) cells. We established a radioactive assay and demonstrated reduced cellular uptake of sulfate in SLC26A2 knockout cells.In the first part of this project, we aim to further characterize sulfate transport in OK cells regarding the transporters involved, their localization and interplay, as well as effects of various other ions on sulfate transport, with a focus on SLC26A2.Global Slc26a2 knockout is lethal in mice. We thus propose to generate a proximal tubule-specific knockout of Slc26a2 to define the role of Slc26a2 in renal sulfate transport. We will perform transport assays and compare sulfate levels in blood and urine between knockout mice and controls.Finally, we aim to study protein domains that determine SLC26A2 trafficking and apical membrane insertion. We will study the effect of dietary sulfate on Slc26a2 expression in the mouse model.We hope that these studies will form a basis for future translational studies on SLC26A2.

硫酸盐对正常细胞功能至关重要。例如，它是蛋白聚糖硫酸化所必需的，蛋白聚糖是有助于正常软骨结构的分子。软骨硫酸盐转运蛋白（SLC 26 A2）的功能缺失突变是一组常染色体隐性软骨发育不良的原因。在这些疾病中，软骨细胞吸收硫酸盐受损导致蛋白聚糖硫酸化减少。然而，SLC 26 A2不仅在软骨中表达，而且在其他组织中表达，例如肾脏。在那里，它特异性地定位于近端肾小管细胞的顶端（管腔）刷状缘膜。已知硫酸盐在肾脏，特别是近端小管中被自由过滤和广泛重吸收，但SLC 26 A2在肾脏硫酸盐转运中的确切功能尚未被探索。在初步研究中，使用CRISPR/Cas9系统，我们在近端小管OK（负鼠肾）细胞的细胞模型中敲除了SLC 26 A2。我们建立了一种放射性分析方法，证明了SLC 26 A2基因敲除细胞对硫酸盐的摄取减少。在本项目的第一部分，我们的目标是进一步表征OK细胞中硫酸盐转运的相关转运蛋白，它们的定位和相互作用，以及各种其他离子对硫酸盐转运的影响，重点是SLC 26 A2。因此，我们建议产生近端小管特异性敲除Slc 26 a2，以确定Slc 26 a2在肾脏硫酸盐转运中的作用。我们将进行转运试验，并比较敲除小鼠和对照小鼠血液和尿液中的硫酸盐水平。最后，我们的目标是研究决定SLC 26 A2运输和顶端膜插入的蛋白质结构域。我们将在小鼠模型中研究膳食硫酸盐对SLC 26 A2表达的影响，希望这些研究能为将来SLC 26 A2的翻译研究奠定基础。