Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity
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 CD...
| Autores: | , , , , , , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Fecha de publicación: | 2019 |
| País: | España |
| Institución: | Instituto de Salud Carlos III (ISCIII) |
| Repositorio: | Repisalud |
| Idioma: | inglés |
| OAI Identifier: | oai:repisalud.isciii.es:20.500.12105/9319 |
| Acceso en línea: | http://hdl.handle.net/20.500.12105/9319 |
| Access Level: | acceso abierto |
| Palabra clave: | Animals Cell Differentiation Cell Line, Tumor Epidermal Growth Factor Gene Expression Profiling Gene Ontology Gene Regulatory Networks Humans Integrin alpha6 Mice, 129 Strain Mice, Inbred C57BL Mice, Nude Mice, Transgenic Organoids Receptors, Notch Single-Cell Analysis Stem Cells Urothelium |
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Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacitySantos, Catarina PLapi, EleonoraMartínez de Villarreal, JaimeÁlvaro-Espinosa, LauraFernández-Barral, AsunciónBarbáchano, AntonioDominguez, OrlandoLaughney, Ashley MMegias Vazquez, DiegoMuñoz, AlbertoReal Arribas, FranciscoAnimalsCell DifferentiationCell Line, TumorEpidermal Growth FactorGene Expression ProfilingGene OntologyGene Regulatory NetworksHumansIntegrin alpha6Mice, 129 StrainMice, Inbred C57BLMice, NudeMice, TransgenicOrganoidsReceptors, NotchSingle-Cell AnalysisStem CellsUrotheliumUnderstanding 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.Nature Publishing GroupMinisterio de Economía y Competitividad (España)Centro de Investigación Biomédica en Red - CIBERONC (Cáncer)Asociación Española Contra el Cáncer20202020-03-2420192019-09-2720192019-09-27journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfvideo/quicktimeapplication/pdfapplication/vnd.openxmlformats-officedocument.spreadsheetml.sheetapplication/vnd.openxmlformats-officedocument.spreadsheetml.sheetvideo/quicktimeapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/20.500.12105/9319reponame:Repisaludinstname:Instituto de Salud Carlos III (ISCIII)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Atribución-NoComercial-CompartirIgual 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/openAccessoai:repisalud.isciii.es:20.500.12105/93192026-06-12T12:43:37Z |
| dc.title.none.fl_str_mv |
Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity |
| title |
Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity |
| spellingShingle |
Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity Santos, Catarina P Animals Cell Differentiation Cell Line, Tumor Epidermal Growth Factor Gene Expression Profiling Gene Ontology Gene Regulatory Networks Humans Integrin alpha6 Mice, 129 Strain Mice, Inbred C57BL Mice, Nude Mice, Transgenic Organoids Receptors, Notch Single-Cell Analysis Stem Cells Urothelium |
| title_short |
Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity |
| title_full |
Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity |
| title_fullStr |
Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity |
| title_full_unstemmed |
Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity |
| title_sort |
Urothelial organoids originating from Cd49fhigh mouse stem cells display Notch-dependent differentiation capacity |
| dc.creator.none.fl_str_mv |
Santos, Catarina P Lapi, Eleonora Martínez de Villarreal, Jaime Álvaro-Espinosa, Laura Fernández-Barral, Asunción Barbáchano, Antonio Dominguez, Orlando Laughney, Ashley M Megias Vazquez, Diego Muñoz, Alberto Real Arribas, Francisco |
| author |
Santos, Catarina P |
| author_facet |
Santos, Catarina P Lapi, Eleonora Martínez de Villarreal, Jaime Álvaro-Espinosa, Laura Fernández-Barral, Asunción Barbáchano, Antonio Dominguez, Orlando Laughney, Ashley M Megias Vazquez, Diego Muñoz, Alberto Real Arribas, Francisco |
| author_role |
author |
| author2 |
Lapi, Eleonora Martínez de Villarreal, Jaime Álvaro-Espinosa, Laura Fernández-Barral, Asunción Barbáchano, Antonio Dominguez, Orlando Laughney, Ashley M Megias Vazquez, Diego Muñoz, Alberto Real Arribas, Francisco |
| author2_role |
author author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
Ministerio de Economía y Competitividad (España) Centro de Investigación Biomédica en Red - CIBERONC (Cáncer) Asociación Española Contra el Cáncer |
| dc.subject.none.fl_str_mv |
Animals Cell Differentiation Cell Line, Tumor Epidermal Growth Factor Gene Expression Profiling Gene Ontology Gene Regulatory Networks Humans Integrin alpha6 Mice, 129 Strain Mice, Inbred C57BL Mice, Nude Mice, Transgenic Organoids Receptors, Notch Single-Cell Analysis Stem Cells Urothelium |
| topic |
Animals Cell Differentiation Cell Line, Tumor Epidermal Growth Factor Gene Expression Profiling Gene Ontology Gene Regulatory Networks Humans Integrin alpha6 Mice, 129 Strain Mice, Inbred C57BL Mice, Nude Mice, Transgenic Organoids Receptors, Notch Single-Cell Analysis Stem Cells Urothelium |
| description |
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. |
| publishDate |
2019 |
| dc.date.none.fl_str_mv |
2019 2019-09-27 2019 2019-09-27 2020 2020-03-24 |
| dc.type.none.fl_str_mv |
journal article http://purl.org/coar/resource_type/c_6501 VoR http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| dc.type.openaire.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/20.500.12105/9319 |
| url |
http://hdl.handle.net/20.500.12105/9319 |
| dc.language.none.fl_str_mv |
Inglés eng |
| language_invalid_str_mv |
Inglés |
| language |
eng |
| dc.rights.none.fl_str_mv |
open access http://purl.org/coar/access_right/c_abf2 Atribución-NoComercial-CompartirIgual 4.0 Internacional http://creativecommons.org/licenses/by-nc-sa/4.0/ |
| dc.rights.openaire.fl_str_mv |
info:eu-repo/semantics/openAccess |
| rights_invalid_str_mv |
open access http://purl.org/coar/access_right/c_abf2 Atribución-NoComercial-CompartirIgual 4.0 Internacional http://creativecommons.org/licenses/by-nc-sa/4.0/ |
| eu_rights_str_mv |
openAccess |
| dc.format.none.fl_str_mv |
application/pdf video/quicktime application/pdf application/vnd.openxmlformats-officedocument.spreadsheetml.sheet application/vnd.openxmlformats-officedocument.spreadsheetml.sheet video/quicktime application/pdf application/pdf application/pdf |
| dc.publisher.none.fl_str_mv |
Nature Publishing Group |
| publisher.none.fl_str_mv |
Nature Publishing Group |
| dc.source.none.fl_str_mv |
reponame:Repisalud instname:Instituto de Salud Carlos III (ISCIII) |
| instname_str |
Instituto de Salud Carlos III (ISCIII) |
| reponame_str |
Repisalud |
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Repisalud |
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1869407050557030400 |
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15,811543 |