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Organoid cystogenesis reveals a critical role of microenvironment in human polycystic kidney disease

Nature Materials volume 16, pages 1112–1119 (2017)Cite this article

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Abstract

Polycystic kidney disease (PKD) is a life-threatening disorder, commonly caused by defects in polycystin-1 (PC1) or polycystin-2 (PC2), in which tubular epithelia form fluid-filled cysts1,2. A major barrier to understanding PKD is the absence of human cellular models that accurately and efficiently recapitulate cystogenesis3,4. Previously, we have generated a genetic model of PKD using human pluripotent stem cells and derived kidney organoids5,6. Here we show that systematic substitution of physical components can dramatically increase or decrease cyst formation, unveiling a critical role for microenvironment in PKD. Removal of adherent cues increases cystogenesis 10-fold, producing cysts phenotypically resembling PKD that expand massively to 1-centimetre diameters. Removal of stroma enables outgrowth of PKD cell lines, which exhibit defects in PC1 expression and collagen compaction. Cyclic adenosine monophosphate (cAMP), when added, induces cysts in both PKD organoids and controls. These biomaterials establish a highly efficient model of PKD cystogenesis that directly implicates the microenvironment at the earliest stages of the disease.

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Figure 1: Removal of adherent cues establishes a highly efficient model of PKD cystogenesis.
Figure 2: Organoid PKD cysts phenotypically resemble PKD patient cysts.
Figure 3: PKD organoid cysts arise from hyperproliferative KTECs.
Figure 4: Outgrowth of PKD cell lines reveals a critical deficiency in PC1 expression.
Figure 5: Organoids remodel their matrix microenvironment in a PKD-dependent manner.

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Acknowledgements

The authors thank S. Shankland, E. Kelly, D. Beier, H. Ruohola-Baker and C. Murry (UW), and J. Bonventre, J. Zhou and T. Steinman (Harvard Medical School) for helpful discussions. We thank D. Hailey (ISCRM Garvey Imaging Core) and E. Parker (UW Vision Core Lab) for microscopy support. The Laboratory of Developmental Biology was supported by NIH Award Number R24HD000836 (NICHD). Studies were supported by an NIH Career Development Award K01DK102826 (NIDDK), PKD Foundation Research Award, National Kidney Foundation Young Investigator Grant, and American Society of Nephrology Carl W. Gottschalk Research Scholar Award (B.S.F.). The work was also supported by start-up funds from the University of Washington, NIH K25HL135432 (H.F.), NIH UH3TR000504 and UG3TR002158 (J.H.), and an unrestricted gift from the Northwest Kidney Centers to the Kidney Research Institute.

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Authors and Affiliations

  1. Division of Nephrology, University of Washington School of Medicine, Seattle, Washington, 98109, USA

    Nelly M. Cruz, Stefan M. Czerniecki, Ramila E. Gulieva, Angela J. Churchill, Yong Kyun Kim, Kosuke Winston, Linh M. Tran, Marco A. Diaz, Jonathan Himmelfarb & Benjamin S. Freedman

  2. Kidney Research Institute, University of Washington School of Medicine, Seattle, Washington, 98109, USA

    Nelly M. Cruz, Stefan M. Czerniecki, Ramila E. Gulieva, Angela J. Churchill, Yong Kyun Kim, Kosuke Winston, Linh M. Tran, Marco A. Diaz, Jonathan Himmelfarb & Benjamin S. Freedman

  3. Institute for Stem Cell and Regenerative Medicine, University of Washington School of Medicine, Seattle, Washington, 98109, USA

    Nelly M. Cruz, Stefan M. Czerniecki, Ramila E. Gulieva, Angela J. Churchill, Yong Kyun Kim, Kosuke Winston, Linh M. Tran, Marco A. Diaz, Hongxia Fu & Benjamin S. Freedman

  4. Department of Medicine, University of Washington School of Medicine, Seattle, Washington, 98109, USA

    Nelly M. Cruz, Stefan M. Czerniecki, Ramila E. Gulieva, Angela J. Churchill, Yong Kyun Kim, Kosuke Winston, Linh M. Tran, Marco A. Diaz, Hongxia Fu, Jonathan Himmelfarb & Benjamin S. Freedman

  5. Division of Nephrology, University Health Network, Toronto, Ontario M5G2N2, Canada

    Xuewen Song & York Pei

  6. University of Toronto, Toronto, Ontario M5G2N2, Canada

    Xuewen Song & York Pei

  7. Division of Hematology, Department of Medicine, University of Washington School of Medicine, Seattle, Washington, 98109, USA

    Hongxia Fu

  8. Department of Bioengineering, University of Washington School of Medicine, Seattle, Washington, 98109, USA

    Hongxia Fu

  9. Department of Pathology, University of Washington School of Medicine, Seattle, Washington, 98105, USA

    Laura S. Finn

  10. Department of Laboratories, Seattle Children’s Hospital, Seattle, Washington, 98105, USA

    Laura S. Finn

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  1. Nelly M. Cruz
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Contributions

N.M.C., S.M.C., R.E.G., A.J.C., Y.K.K., K.W., L.M.T., M.A.D. and B.S.F. performed experiments and analysis of hPSCs and kidney organoids. X.S. and Y.P. performed global gene analysis of kidney organoid cysts. L.S.F., M.A.D. and B.S.F. analysed tissues from PKD patients. N.M.C., X.S., S.M.C., H.F., L.S.F., Y.P., J.H. and B.S.F. contributed to experimental design and analysis. B.S.F. and N.M.C. wrote the first draft of the manuscript. All authors revised the manuscript.

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Correspondence to Benjamin S. Freedman.

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Competing interests

B.S.F. is an inventor on patent applications for kidney differentiation and disease modelling from human pluripotent stem cells (PCT/US14/34031, PCT/US16/50271).

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Cruz, N., Song, X., Czerniecki, S. et al. Organoid cystogenesis reveals a critical role of microenvironment in human polycystic kidney disease. Nature Mater 16, 1112–1119 (2017). https://doi.org/10.1038/nmat4994

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