Urothelial organoids originating from Cd49f
Animals
Cell Differentiation
/ drug effects
Cell Line, Tumor
Epidermal Growth Factor
/ pharmacology
Gene Expression Profiling
/ methods
Gene Ontology
Gene Regulatory Networks
Humans
Integrin alpha6
/ genetics
Mice, 129 Strain
Mice, Inbred C57BL
Mice, Nude
Mice, Transgenic
Organoids
/ cytology
Receptors, Notch
/ genetics
Single-Cell Analysis
/ methods
Stem Cells
/ cytology
Urothelium
/ cytology
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
27 09 2019
27 09 2019
Historique:
received:
04
08
2018
accepted:
02
09
2019
entrez:
29
9
2019
pubmed:
29
9
2019
medline:
14
1
2020
Statut:
epublish
Résumé
Understanding urothelial stem cell biology and differentiation has been limited by the lack of methods for their unlimited propagation. Here, we establish mouse urothelial organoids that can be maintained uninterruptedly for >1 year. Organoid growth is dependent on EGF and Wnt activators. High CD49f/ITGA6 expression features a subpopulation of organoid-forming cells expressing basal markers. Upon differentiation, multilayered organoids undergo reduced proliferation, decreased cell layer number, urothelial program activation, and acquisition of barrier function. Pharmacological modulation of PPARγ and EGFR promotes differentiation. RNA sequencing highlighted genesets enriched in proliferative organoids (i.e. ribosome) and transcriptional networks involved in differentiation, including expression of Wnt ligands and Notch components. Single-cell RNA sequencing (scRNA-Seq) analysis of the organoids revealed five clusters with distinct gene expression profiles. Together, with the use of γ-secretase inhibitors and scRNA-Seq, confirms that Notch signaling is required for differentiation. Urothelial organoids provide a powerful tool to study cell regeneration and differentiation.
Identifiants
pubmed: 31562298
doi: 10.1038/s41467-019-12307-1
pii: 10.1038/s41467-019-12307-1
pmc: PMC6764959
doi:
Substances chimiques
Integrin alpha6
0
Receptors, Notch
0
Epidermal Growth Factor
62229-50-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
4407Références
Kidney Int. 2005 Sep;68(3):1032-40
pubmed: 16105033
Nat Methods. 2017 May;14(5):483-486
pubmed: 28346451
Br J Urol. 1969 Dec;41(6):710-3
pubmed: 5359494
Nat Biotechnol. 2018 Jun;36(5):421-427
pubmed: 29608177
Lab Invest. 2004 Apr;84(4):465-78
pubmed: 14968126
Nat Commun. 2016 Jun 20;7:11914
pubmed: 27320313
Nature. 2011 Apr 7;472(7341):110-4
pubmed: 21389986
Am J Pathol. 2004 May;164(5):1789-98
pubmed: 15111325
Sci Transl Med. 2014 Jul 9;6(244):244ra91
pubmed: 25009231
Nat Med. 2011 Sep 04;17(10):1225-7
pubmed: 21892181
Nat Rev Urol. 2012 Oct;9(10):583-94
pubmed: 22890301
Dev Cell. 2013 Sep 16;26(5):469-482
pubmed: 23993789
Am J Physiol Renal Physiol. 2004 Aug;287(2):F305-18
pubmed: 15068973
Cell. 2015 Jan 15;160(1-2):299-312
pubmed: 25533785
Bladder Cancer. 2016 Jan 7;2(1):37-47
pubmed: 27376123
Proc Natl Acad Sci U S A. 2019 Mar 5;116(10):4567-4574
pubmed: 30787188
Cell. 2018 Apr 5;173(2):515-528.e17
pubmed: 29625057
Dev Biol. 2011 Oct 1;358(1):79-90
pubmed: 21803035
Nature. 2009 May 14;459(7244):262-5
pubmed: 19329995
Nat Cell Biol. 2016 Mar;18(3):246-54
pubmed: 26911908
Cell Death Differ. 2009 Jan;16(1):103-14
pubmed: 18688264
J Pathol. 2017 May;242(1):113-125
pubmed: 28195647
J Clin Invest. 2015 Feb;125(2):824-30
pubmed: 25574842
Cell. 2016 Jun 16;165(7):1586-1597
pubmed: 27315476
Oncogene. 2011 Jan 13;30(2):178-89
pubmed: 20818428
Traffic. 2004 Mar;5(3):117-28
pubmed: 15086788
Bladder Cancer. 2018 Apr 26;4(2):215-226
pubmed: 29732392
Eur Urol. 2011 Jul;60(1):141-9
pubmed: 21353380
Am J Pathol. 2015 May;185(5):1385-95
pubmed: 25907831
Cell. 2015 Jan 15;160(1-2):324-38
pubmed: 25557080
Virchows Arch B Cell Pathol Incl Mol Pathol. 1986;51(1):65-70
pubmed: 2871663
Proc Natl Acad Sci U S A. 2012 Feb 7;109(6):2078-83
pubmed: 22308455
Nat Genet. 2013 Dec;45(12):1464-9
pubmed: 24121791
Nat Methods. 2014 Jan;11(1):106-12
pubmed: 24292484
J Cell Sci. 2004 Apr 15;117(Pt 10):2029-36
pubmed: 15054105
Nat Biotechnol. 2018 Jun;36(5):411-420
pubmed: 29608179
Nat Biotechnol. 2018 Dec 03;:
pubmed: 30531897
Cell. 2017 Oct 19;171(3):540-556.e25
pubmed: 28988769
Dev Cell. 2016 Sep 26;38(6):590-600
pubmed: 27676432
Genome Biol. 2016 Feb 17;17:29
pubmed: 26887813
Proc Natl Acad Sci U S A. 2012 Mar 27;109(13):4974-9
pubmed: 22416122
Hematol Oncol Clin North Am. 2015 Apr;29(2):177-89, vii
pubmed: 25836927
Cell. 1995 Jan 13;80(1):83-93
pubmed: 7813021
Nat Med. 2014 Oct;20(10):1199-205
pubmed: 25194568
Cell Tissue Res. 2012 Jun;348(3):589-600
pubmed: 22427119
BMC Cancer. 2014 Aug 29;14:628
pubmed: 25167871
Cell. 2019 Jun 13;177(7):1888-1902.e21
pubmed: 31178118
Proc Natl Acad Sci U S A. 2009 Aug 18;106(33):14016-21
pubmed: 19666525
Cell. 2014 Sep 25;159(1):163-175
pubmed: 25201529
Am J Pathol. 1998 Aug;153(2):349-54
pubmed: 9708794
J Biol Chem. 1990 Nov 5;265(31):19170-9
pubmed: 2229070
J Cell Biol. 1965 Jul;26(1):25-48
pubmed: 5859020
Dis Model Mech. 2011 Jul;4(4):548-55
pubmed: 21504907
J Biol Chem. 2002 Aug 23;277(34):30998-1004
pubmed: 12055200