DepartmentKidney Development
Publication date4-Aug-2020

Shohei Kuraoka, Shunsuke Tanigawa, Atsuhiro Taguchi, Akitsu Hotta, Hitoshi Nakazato, Kenji Osafune, Akio Kobayashi, Ryuichi Nishinakamura. PKD1-dependent renal cystogenesis in human induced pluripotent stem cell-derived ureteric bud/collecting duct organoids. Journal of the American Society of Nephrology, in press (2020)


Background Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary kidney disease leading to renal failure, wherein multiple cysts are formed in renal tubules and collecting ducts derived from distinct precursors: the nephron progenitor and ureteric bud (UB), respectively. Recent progress in induced pluripotent stem cell (iPSC) biology has enabled cyst formation in nephron progenitor-derived human kidney organoids lacking PKD1 and PKD2, the major causative genes for ADPKD. However, cyst formation in UB organoids has not been achieved, despite the prevalence of collecting duct cysts in ADPKD patients.


Methods We deleted PKD1 in human iPSCs using CRISPR-Cas9 technology and differentially induced the cells toward nephron or UB organoids. We then investigated cyst formation in both types of kidney organoid. We also examined cyst formation in UB organoids generated from ADPKD patient-derived iPSCs.


Results As observed in nephron organoids, cysts were formed in UB organoids with homozygous PKD1 mutations upon cAMP stimulation, and to a lesser extent, in heterozygous mutant organoids. Furthermore, UB organoids generated from ADPKD patient-derived iPSCs with a heterozygous missense mutation showed cystogenesis upon cAMP stimulation.


Conclusions We demonstrated cyst formation in PKD1 mutant UB organoids, as well as those derived from an ADPKD patient. Our findings will serve as a valuable basis for elucidating the mechanisms of ADPKD.



Figure. PKD1 mutant UB organoids exhibit cystogenesis upon forskolin treatment.

(A) Representative bright-field images of control UB organoids with forskolin treatment at day 30. Scale bar, 200 μm. (B) Representative bright-field images of PKD1 mutant UB organoids with forskolin treatment at day 30. Multiple cyst formation was observed. Scale bar, 200 μm.  (C) Confocal images of immunofluorescence in PKD1 mutant UB organoids at day 30. The Cyst epithelia, as well as normal epithelia in induced UB organoids (arrow), expressed DBA (green) and KRT19 (red), which are the markers of collecting ducts. The asterisks indicate the lumens of cysts. Scale bar, 100 μm.