ESR 15: Generation of ADPKD in vitro models for genotype-phenotype correlation and drug discovery

Supervisor: Ariela Benigni

PhD Student: Alexandru Scarlat

Short Summary

Autosomal dominant polycystic kidney disease (ADPKD) is the most common renal genetic disease, affecting 1 in 1,000 individuals. It is characterised by the growth of multiple fluid-filled cysts, leading to renal failure. ADPKD is caused by mutations in the genes PKD1 and PKD2, encoding the proteins polycystin 1 and polycystin 2 respectively. Only one drug has been approved so far, tolvaptan, however its use is limited by variability in patient response and side effects. Novel models are therefore required to study ADPKD initiation and progression, as well as a platform for drug screening. To address this, we will develop a novel tridimensional model of collecting ducts, the main site of cyst formation, and use healthy as well as PKD1 and PKD2 mutated induced pluripotent stem cells (iPSCs) to study ADPKD. Patient-derived iPSCs will also be generated, and our model will then be used for drug screening to identify effective treatments for patients.

Scientific strategy


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.

Expected Results:

  • 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.