The project aims to develop a new platform for 3D collecting duct tubules derived from iPSCs in order to study ADPKD initiation and progression. We aim to understand the molecular basis of cyst formation in PKD1 and PKD2 mutant cell lines, as well as the roles and implications of these mutated genes. This knowledge will then inform the drug screening process, which will allow the identification of novel therapeutics.
A differentiation protocol for the formation of collecting ducts from iPSCs will be developed on a 3D platform in order to study cystogenesis mechanisms. Next generation sequencing (NGS) technology will then be used to study altered signalling pathways and CRISPR-Cas9 will be used to study the role of PKD1 and PKD2 in patient-derived iPSCs. Finally, drug screening will be performed on the 3D tubule platform, based on the identified altered pathways.
- Development of a 3D collecting duct platform and identification of modified pathways in PKD1 and PKD2 mutant tissues.
- Generation of iPSCs from ADPKD patients and correction of the disease-causing mutation with CRISPR-Cas9.
- Identification of novel targets in ADPKD from drug screening on the 3D tubule platform.