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Kidney Week

Abstract: TH-PO019

Modeling of Tacrolimus Nephrotoxicity Using Kidney Organoids Derived from Human iPS Cells

Session Information

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Kim, Jinwon, Catholic university of Korea, Seoul, Korea (the Republic of)
  • Freedman, Benjamin S., University of Washington, Seattle, Washington, United States
  • Kim, Yong Kyun, Catholic University of Korea, Seoul, Korea (the Republic of)
Background

Tacrolimus, a calcineurin inhibitor, was clinically used as an immunosuppressive agent in organ transplantation or glomerulonephritis. Despite the therapeutic benefits, tacrolimus’s use is limited due to its nephrotoxicity. To reduce tacrolimus nephrotoxicity, the effective experimental models are essential. Recently, we and others have established protocols for the generation of kidney organoids from human pluripotent stem cells, containing nephron-like structures with podocytes, proximal tubules, and distal tubules. Here, we recapitulated tacrolimus nephrotoxicity using kidney organoids and investigated its pathogenic mechanism.

Methods

Kidney organoid differentiated from the CMC11 iPSC cell line (human male donor). Kidney organoids were re-seeded in 96-well plates and tacrolimus was treated at doses of 0 μM, 30 μM, 60 μM, or 120 μM for 24 h.

Results

The size of kidney organoids decreased at dose-dependent manner. Cell viability assessed by CCK-8 assay and live/dead cell staining decreased at dose-dependent manner. Proximal tubular cells as well as distal tubular cells were decreased according to the concentration of tacrolimus. Podocyte loss and injured podocytes with unplolarized and diffuse pattern of ZO-1 tracks were observed after treatment of tacrolimus. Ultrastructural analyses showed the vacuoles throughout the cytoplasm of tubule-like structures, which were similar to those of human tacrolimus nephrotoxicity. Autophagic activity was enhanced after treatment with tacrolimus in kidney organoids, which were similar patterns in mouse model of tacrolimus nephrotoxicity. Rapamycin, as an autophagy inducer, attenuated cell death in kidney organoids model of tacrolimus nephrotoxicity, whereas 3-methyladenine, as an autophagy inhibitor, accelerated cellular toxicity.

Conclusion

Our data suggest that human iPSC-derived kidney organoids can recapitulate tacrolimus nephrotoxicity and serve the effective in vitro model to investigate its pathogenic pathway.