TY  - GEN
AV  - public
A1  - Pandey, Gunjan
CY  - Heidelberg
TI  - Development of An Automated High-Content Screening Platform for Identification of Cystic Kidney Disease Modifying Substances in Zebrafish Model of Human Cystic Kidney Disease
UR  - https://archiv.ub.uni-heidelberg.de/volltextserver/30328/
Y1  - 2022///
ID  - heidok30328
N2  - In this work, I describe technical and biological aspects of a High-Content Screening experiment conducted to identify pharmacological modifiers of human cystic kidney disease in zebrafish larvae. Human cystic nephropathy was mimicked in zebrafish larvae with the help of morpholino-mediated knockdown of the ift172 gene. IFT172 is part of the ciliary intra flagellar transport complex; its deficiency impairs the function of the primary cilia, resulting in impaired urine flow and the development of fluid filled cysts. An automated pipeline was developed to optimise large scale sample handling, image acquisition, quantification and analysis. Dedicated hardware and software tools were developed to solve issues associated with the spatial orientation of larvae in microtitre plates, automated
Image capture, data pre-processing and quantification. After successful implementation of the high-content screening platform, I screened 1,280 known small-molecular pharmacological compounds from the Prestwick library in 12 larvae per compound. The initial screen yielded 60 candidate compounds with potential cyst suppressive activity. The hits were further validated in dose response studies, which led to the final confirmation of 17 target compounds. These compounds share several interesting putative modes of action, including estrogen antagonism, enkephalinase and proton pump inhibition, anti-tyrosinase activity, calcium channel regulation, retionoic acid receptor interactions and topoisomerase blockade. In conclusion, the establishment and execution of an automated High-Content Screening in a transgenic zebrafish model yielded several promising novel pharmacological targets that can be further explored with regards to their cyst-suppressing activity and suitability for use in human
cystic kidney diseases.
ER  -