Development of A Cell-Based Assay to Identify Small Molecule Inhibitors of FGF23 Signaling

Diener S, Schorpp K, Strom T, Hadian K, Lorenz-Depiereux B
ASSAY and Drug Development Technologies - vol. 13 476-487 (2015)

ASSAY and Drug Development Technologies

Abstract Fibroblast growth factor 23 (FGF23) is a bone-derived endocrine key regulator of phosphate homeostasis. It inhibits renal tubular phosphate reabsorption by activating receptor complexes composed of FGF receptor 1c (FGFR1c) and the co-receptor Klotho. As a major signaling pathway mitogen-activated protein kinase (MAPK) pathway is employed. In this study, we established an FGF23-inducible cell model by stably expressing human Klotho in HEK293 cells (HEK293-KL cells) containing endogenous FGF receptors. To identify novel small molecule compounds that modulate FGF23/FGFR1c/Klotho signaling, we developed and optimized a cell-based assay that is suited for high-throughput screening. The assay monitors the phosphorylation of endogenous extracellular signal-regulated kinase 1 and 2 in cellular lysates of HEK293-KL cells after induction with FGF23. This cell-based assay was highly robust (Z′ factor >0.5) and the induction of the system is strictly dependent on the presence of FGF23. The inhibitor response…

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