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Abstract: PO1546

Generation of Collecting Duct Kidney Organoids from Human Induced Pluripotent Stem Cell

Session Information

Category: Genetic Diseases of the Kidneys

  • 1001 Genetic Diseases of the Kidneys: Cystic

Authors

  • Kyrychenko, Sergii, Goldfinch Bio Inc, Cambridge, Massachusetts, United States
  • Fast, Eva, Goldfinch Bio Inc, Cambridge, Massachusetts, United States
  • Fanelli, Alyssa, Goldfinch Bio Inc, Cambridge, Massachusetts, United States
  • Bennett, Henrietta W., Goldfinch Bio Inc, Cambridge, Massachusetts, United States
  • Mundel, Peter H., Goldfinch Bio Inc, Cambridge, Massachusetts, United States
  • Reilly, John F., Goldfinch Bio Inc, Cambridge, Massachusetts, United States
  • Westerling-Bui, Amy Duyen, Goldfinch Bio Inc, Cambridge, Massachusetts, United States
Background

Polycystic kidney disease (PKD) is one of the most common human genetic disorders without effective therapy. During its progression, fluid-filled cysts displace normal collecting duct (CD) tubules causing end-stage renal failure. The lack of disease-relevant in vitro models of PKD has hampered early drug discovery and needs more efficient and robust tools.

Methods

Here we modified a previously published protocol [1] and established a high-throughput and highly efficient method for the generation of CD kidney organoids from human induced pluripotent stem cells (hiPSC). We employed a dynamic modulation of cell signaling pathways in combination with 3D extracellular matrix support to induce CXCR4+/cKit+ ureteric bud (UB) cell progenitors and further UB branching.

Results

The UB gives rise to renal collecting ducts and the lower urinary tract. We observed the development of UB-like cytoarchitecture including, bifurcated ureteric tip expressing specific markers (RET, WNT9B, HOXB7). Using single-cell RNA sequencing (scRNAseq) we identified two major cell populations in differentiated CD organoids – collecting duct cells and stromal cells. CD cells express typical markers of UB trunk (CK19), the ureteric epithelium (CDH1, CK8), as well as mature markers (AQP2, CALB1, MUC1) including principal (AQP3) and intercalated cells (AQP5). Moreover, we identified cilia formation on the inner surface of the luminal cavity of CD tubules which mimics normal kidney development. Using pharmacological approaches, we were able to induce cysts formation in response to forskolin and cholera toxin treatment, thus, simulating the abnormal CD response to excess cAMP in PKD or normal rodent embryonic kidneys [2].

Conclusion

In conclusion, we provide a robust and highly efficient method for collecting duct marker expressing organoids that may contribute to elucidating the mechanisms of kidney development, disease modeling of the lower urinary tract (polycystic kidney disease), and drug discovery.

1. Taguchi A, Nishinakamura R. (2017) Higher-order kidney organogenesis from pluripotent stem cells. Cell Ste Cell. 21: 730-746
2. Magenheimer BS et al. (2006). J Am Soc Nephrol. 17(12): 3424-3437

Funding

  • Commercial Support –