Abstract: SA-PO008
New Translational Screening Assay with Phenotypic and Transcriptomic Readouts Using Induced Pluripotent Stem Cell (iPSC)-Derived Cells and Organoids from a Nephrotic Syndrome Patient
Session Information
- Bioengineering: Modeling, Diagnosis, Therapy
November 04, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Bioengineering
- 400 Bioengineering
Authors
- Xian, Wenying, Evotec SE, Hamburg, Hamburg, Germany
- Gratz, Rena, Evotec SE, Hamburg, Hamburg, Germany
- Drössler, Jana, Evotec SE, Hamburg, Hamburg, Germany
- Ercetin, Evrim, Evotec SE, Hamburg, Hamburg, Germany
- Nishinakamura, Ryuichi, Kumamoto Daigaku, Kumamoto, Kumamoto, Japan
- Guhl, Anna, Evotec SE, Hamburg, Hamburg, Germany
- Andag, Uwe, Evotec SE, Hamburg, Hamburg, Germany
- Radresa, Olivier, Evotec SE, Hamburg, Hamburg, Germany
- Schwarz, Nele, Evotec SE, Hamburg, Hamburg, Germany
- Naujock, Maximilian, Evotec SE, Hamburg, Hamburg, Germany
Group or Team Name
- iPSC META.
Background
Professor Nishinakamura’s group previously generated pluripotent stem cell (iPSC)-derived kidney organoids from a patient suffering from a congenital nephrotic syndrome caused by the mutations in the nephrin gene (c.G1379A, c.2515delC in NPHS1). We further developed this system into a robust, reproducible, and high-throughput organoid assay useful for the progression of drug discovery programs.
Methods
To enable readouts in 384w, we scaled up the production of iPSC-podocytes and iPSC-kidney organoids. We also developed and applied a 3D whole mount immunostaining readout to quantify the phenotypic signature of iPSC-derived podocytes and kidney organoids. Bulk ScreenSeq™ and single-nucleus sequencing was applied to characterize the transcriptomic profiles of both systems.
Results
A specific lack of nephrin phosphorylation was confirmed via automated western blotting. Besides, our kidney organoids completely lacked nephrin signals in synaptopodin-positive regions of glomerular structures. We further quantified the general nephrin deficit with a new script-based whole mount imaging analysis. Finally, by comparing transcriptomic disease signatures, we showed the translational relevance of these organoids for drug discovery in the field of podocytopathies.
Conclusion
We successfully developed and characterized a patient-derived kidney organoid assay and made it available in a high-throughput format. This translational system will prove useful in the identification and validation of candidate targets and for the progression of new kidney therapeutics.