GDNF drives rapid tubule morphogenesis in a novel 3D in vitro model for ADPKD

Eryn E. Dixon, Demetrios S. Maxim, Victoria L. Halperin Kuhns, Allison C. Lane-Harris, Patricia Outeda, Andrew J. Ewald, Terry J. Watnick, Paul A. Welling, Owen M. Woodward

Research output: Contribution to journalArticlepeer-review

Abstract

Cystogenesis is a morphological consequence of numerous genetic diseases of the epithelium. In the kidney, the pathogenic mechanisms underlying the program of altered cell and tubule morphology are obscured by secondary effects of cyst expansion. Here, we developed a new 3D tubuloid system to isolate the rapid changes in protein localization and gene expression that correlate with altered cell and tubule morphology during cyst initiation. Mouse renal tubule fragments were pulsed with a cell differentiation cocktail including glial-derived neurotrophic factor (GDNF) to yield collecting duct-like tubuloid structures with appropriate polarity, primary cilia, and gene expression. Using the 3D tubuloid model with an inducible Pkd2 knockout system allowed the tracking of morphological, protein, and genetic changes during cyst formation. Within hours of inactivation of Pkd2 and loss of polycystin-2, we observed significant progression in tubuloid to cyst morphology that correlated with 35 differentially expressed genes, many related to cell junctions, matrix interactions, and cell morphology previously implicated in cystogenesis.

Original languageEnglish (US)
Article numberjcs249557
JournalJournal of cell science
Volume133
Issue number14
DOIs
StatePublished - Jul 2020

Keywords

  • 3D cell model
  • Collecting duct
  • Epithelia
  • Kidney
  • Polycystic kidney disease
  • Tubulogenesis

ASJC Scopus subject areas

  • Cell Biology

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