@article{b816122c9ec34adc973b817c35cad2b7,
title = "GDNF drives rapid tubule morphogenesis in a novel 3D in vitro model for ADPKD",
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.",
keywords = "3D cell model, Collecting duct, Epithelia, Kidney, Polycystic kidney disease, Tubulogenesis",
author = "Dixon, {Eryn E.} and Maxim, {Demetrios S.} and {Halperin Kuhns}, {Victoria L.} and Lane-Harris, {Allison C.} and Patricia Outeda and Ewald, {Andrew J.} and Watnick, {Terry J.} and Welling, {Paul A.} and Woodward, {Owen M.}",
note = "Funding Information: These experiments were funded by the Baltimore PKD Center Pilot Grant, Baltimore PKD Research and Clinical Core Center National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) 5P30DK090868, the PKD Foundation 229G18a (O.M.W.), National Kidney Foundation (NKF) Mini Grant Award (O.M.W.), National Institute of General Medical Sciences (NIGMS)-sponsored Training Program in Integrative Membrane Biology 5T32GM008181 (E.E.D.), and National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) NRSA F31 DK117579-01 (E.E.D.). Deposited in PMC for immediate release. Funding Information: The authors would like to thank the Baltimore PKD Research and Clinical Core Center for their resources and support for this article, as well as the University of Maryland Greenebaum Comprehensive Cancer Center Flow Cytometry Shared Services for their expertise and help with execution and analysis of the flow cytometry experiments and the Genomics Resource Center at the University of Maryland School of Medicine for their aid with library construction and RNA-sequencing. These experiments were funded by the Baltimore PKD Center Pilot Grant, Baltimore PKD Research and Clinical Core Center National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) 5P30DK090868, the PKD Foundation 229G18a (O.M.W.), National Kidney Foundation (NKF) Mini Grant Award (O.M.W.), National Institute of General Medical Sciences (NIGMS)-sponsored Training Program in Integrative Membrane Biology 5T32GM008181 (E.E.D.), and National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) NRSA F31 DK117579-01 (E.E.D.). Deposited in PMC for immediate release. Publisher Copyright: {\textcopyright} 2020. Published by The Company of Biologists Ltd",
year = "2020",
month = jul,
doi = "10.1242/jcs.249557",
language = "English (US)",
volume = "133",
journal = "The Quarterly journal of microscopical science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "14",
}