Abstract: FR-PO342
Modeling Fabry Disease in Kidney and Heart Organoids From Patient-Derived Human Induced Pluripotent Stem Cells
Session Information
- Genetic Diseases: Models, Mechanisms, Treatments
November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1102 Genetic Diseases of the Kidneys: Non-Cystic
Authors
- Laufer, Sandra D., Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Reinelt, Anna, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Wong, Milagros N., Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Blomberg, Linda, Uniklinik Koln, Koln, Nordrhein-Westfalen, Germany
- Kurschat, Christine E., Uniklinik Koln, Koln, Nordrhein-Westfalen, Germany
- Wanner, Nicola, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Puelles, Victor G., Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Huber, Tobias B., Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Braun, Fabian, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
Background
Fabry disease is an inherited lysosomal storage disorder, caused by mutations in the GLA gene, resulting in a multisystemic disease. Currently existing animal models fail to mirror the complexity of this clinical & molecular phenotype, specifically Fabry nephropathy and cardiomyopathy. This project aims to establish informative human in vitro systems for Fabry disease to advance our molecular understanding by employing the recent advances in human induced pluripotent stem cells (hiPSC) and organoid differentiation
Methods
We collected primary urinary cells (PUCs) of 15 Fabry patients with different mutations (classical, classical and chaperone amenable, late onset, unclear significance). 8 PUC samples were reprogrammed into hiPSC with subsequent creation of respective isogenic control lines by CRISPR Cas9 gene editing. These cell lines were differentiated into kidney and heart organoids as well as engineered heart tissue (EHTs) using published protocols.
Results
All patient-derived hiPSC lines and organoids retained decreased enzyme activity and Gb3 accumulation. Fabry mutations did not impair the differentiation into organoids and EHTs. Kidney organoids depicted the presence of marker proteins for different nephron segments, including glomerular structures. Heart organoids showed contractions and contained different cardiac cell types including cardiomyocytes with sarcomers and Z-Disc formation. EHTs displayed functional abnormalities including decreased contractile force and arrhythmic beating indicative of molecular changes due to GLA deficiency.
Conclusion
This project underlines the advantages of novel complex human in vitro disease modelling to study Fabry disease. Ongoing experiments focus on the detailed structural assessment and single cell analyses of the established systems in the presence and absence of available therapies.
Funding
- Commercial Support –