Abstract: SA-PO342
Modeling Kidney Development, Disease, and Plasticity with Clonal Expandable Nephron Progenitor Cells and Nephron Organoids
Session Information
- Development, Organoids, Vascularized Kidneys, Nephrons, and More
November 04, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 600 Development, Stem Cells, and Regenerative Medicine
Authors
- Huang, Biao, University of Southern California, Los Angeles, California, United States
- Zeng, Zipeng, University of Southern California, Los Angeles, California, United States
- Li, Hui, University of Southern California, Los Angeles, California, United States
- Zhang, Chennan, University of Southern California, Los Angeles, California, United States
- Parvez, Riana, University of Southern California, Los Angeles, California, United States
- Pastor-Soler, Nuria M., University of Southern California, Los Angeles, California, United States
- Hallows, Kenneth R., University of Southern California, Los Angeles, California, United States
- Lindström, Nils, University of Southern California, Los Angeles, California, United States
- McMahon, Andrew P., University of Southern California, Los Angeles, California, United States
- Li, Zhongwei, University of Southern California, Los Angeles, California, United States
Group or Team Name
- Zhongwei Li Lab.
Background
During kidney organogenesis, nephron progenitor cells (NPCs) self-renew and differentiate into nephrons, the functional units of the kidney. Dysregulation of NPC fates underlies a number of congenital kidney diseases while uncontrolled proliferation of NPCs in Wilms tumor is the most prevalent pediatric kidney cancer. Thus, a deeper insight into NPC biology is central to improving an understanding of kidney development, congenital disease and cancer, and to applying developmental insight to regenerating kidney functions.
Methods
A synthetic niche is formulated that allows the in vitro long-term clonal expansion of primary mouse and human NPCs, and induced NPCs (iNPCs) from human pluripotent stem cells. Nephron organoids are generated from cultured NPCs following a chemically-defined differentiation protocol. Genome-wide CRISPR screening is performed in the cultured NPCs to identify novel genes associated with kidney development and disease. Multiplexed CRISPR/Cas9 genome editing in the cultured NPCs generate a rapid, efficient, and scalable organoid model for polycystic kidney disease (PKD), allowing drug screening.
Results
Cultured iNPCs resemble closely primary human NPCs, generating nephron organoids with abundant distal convoluted tubule cells, which are not observed in published kidney organoids. The synthetic niche reprograms differentiated nephron cells into NPC state, recapitulating the plasticity of developing nephron in vivo. Scalability and ease of genome-editing in the cultured NPCs allow for genome-wide CRISPR screening, identifying novel genes associated with kidney development, congenital kidney diseases, and Wilms tumor. Cystic PKD organoid models are generated directly from genome-edited human NPCs in as short as 8 days. Proof-of-concept drug screening identified a drug candidate targeting epigenetic pathway.
Conclusion
These NPC and nephron organoid-based technological platforms have broad applications to kidney development, disease, plasticity, and regeneration.
Funding
- NIDDK Support