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Rapid target validation in a Cas9-inducible hiPSC derived kidney model
Yasaman Shamshirgaran1, Anna Jonebring1, Anna Svensson1
1Translational Genomics, Discovery Sciences, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden.
Scientific Reports
|August 17, 2021
Summary
This study introduces a faster method for creating kidney organoids for disease modeling. It enables efficient genetic editing in induced pluripotent stem cells (iPSCs) to develop accurate models for drug discovery.
Area of Science:
- Stem cell biology
- Genetics
- Regenerative medicine
Background:
- Induced pluripotent stem cells (iPSCs), genome editing, and 3D organoid systems offer new avenues for in vitro human disease models in drug discovery.
- Kidney organoids provide complex cellularity for disease modeling, but genetic manipulation is hindered by the need for laborious clonal cell line generation.
- Current methods for genetically engineering kidney organoids are time-consuming due to the requirement of generating genetically modified clonal lines beforehand.
Purpose of the Study:
- To develop a streamlined methodology for generating genetically engineered kidney organoids.
- To bypass the laborious clonal cell line generation step in creating disease models.
- To establish a scalable platform for drug and genetic screening using engineered kidney organoids.
Main Methods:
- Utilized a doxycycline-inducible Cas9 expressing hiPSC line for direct differentiation of CRISPR-targeted cell pools.
- Employed CRISPR-Cas9 technology for high-efficiency editing in iPSC pool populations.
- Targeted autosomal dominant polycystic kidney disease (ADPKD) genes PKD1 and PKD2 in the engineered iPSCs.
Main Results:
- Achieved over 80% editing efficiency in the iPSC pool population.
- Successfully generated genetically engineered kidney organoids by direct differentiation of edited cell pools.
- Observed cyst-like structures in the tubular compartment of engineered kidney organoids, modeling ADPKD.
- Demonstrated the formation of reliable 3D organoid models of ADPKD.
Conclusions:
- The presented methodology enables efficient genetic manipulation of iPSCs for direct differentiation into kidney organoids.
- This approach significantly reduces the time and labor associated with generating genetically engineered organoid models.
- The developed method provides a versatile platform for rapid target validation and disease modeling, particularly for conditions like ADPKD.
