Analysis of the Role of the Primary Cilium of Renal Epithelial Cells in Fluid Flow Mechanotransduction
Autosomal Dominant Polycystic Kidney Disease (ADPKD) is a hereditary disease that is a leading cause of renal replacement and failure worldwide. The disease has been linked to dysfunctional primary cilia in renal epithelial cells under fluid flow shear stress. Cellular pathways become perturbed due to impaired mechanotransduction, ultimately leading to the formation of renal cysts. This study aimed to determine how mechanotransduction is altered when the primary cilium is removed in order to gain insight into dysregulated mechanotransduction in PKD. Madin-Carby Canine Kidney (MDCK) cells were cultured and plated in two different environments including static and fluid flow shear stress. The primary cilia of the experimental cells were removed using ammonium sulfate in order to emulate dysfunctional cilia in individuals with ADPKD. The control cohort included renal epithelial cells with in-tact primary cilia while the experimental cohort included those in which primary cilia were removed. Both cohorts were exposed to static and fluid flow shear stress environments. Both pathways culminated in an analysis of their biochemical responses through global metabolic profiling. This study has identified ether lipid metabolism to be one significant metabolic pathway that is altered as a result of unregulated mechanotransduction. Additionally, there were many other pathways that corresponded to large numbers of detected metabolites including fatty acid elongation, degradation, and biosynthesis of unsaturated fatty acids. By discovering how this fatty acid synthesis pathway is affected, future therapeutic drug treatments could be created to target these pathways and prevent the progression or development of PKD in susceptible individuals.