Náplní dizrtační práce bude in vitro studium lidského transportomu kyseliny močové na modelu lidských epiteliálních buněk proximálního tubulu.
Preliminary scope of work in English
Uric acid is a primary metabolite of purine degradation in human. It is an important antioxidating substrate and supposed blood pressure regulator. Uric acid is reabsorbed in kidney via epithelial cells of proximal tubule. The most common allelic variants of uric acid transporters (URAT1, GLUT9, ABCG2, OAT1, OAT3 and MRP4) were described in human by genome-wide association studies (GWAS). So far, only functional analysis based on the transport efficiency or kinetics of individual allelic variants were performed However their mutual interactions in in vitro condition using cell culture were not determined yet.. The aim of this project is the study of human uric acid transportome using human kidney epithelial cell culture and CRISPR/Cas9 and siRNA approach to knock-out or downregulate particular uric acid transporters or their allelic variants. On the protein level specific inhibitors of particulate transporters would be employed. The main scientific questions is: “how downregulation or knock-out of selected wt uric acid transporter(s) or their allelic variants affects the uric acid uptake or efflux by epithelial cells via remaining proteins”. To address this question we would determine the transport of radioactive labeled uric acid in apical and basolateral part, perform the kinetic studies and functional studies employing Xenopus laevis oocytes.
- assigned by the advisor