Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine

Autores
Videla Richardson, Guillermo; Furmento, Verónica Alejandra; Garcia, Carolina Paola; Morris Hanon, Olivia; Sevlever, Gustavo; Romorini, Leonardo; Scassa, Maria Elida
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: The essentially unlimited expansion potential and the pluripotency of human embryonic stem cells (hESCs) make them attractive for cell-based therapeutic purposes. Although hESCs can indefinitely proliferate in culture, unlike transformed cancer cells, they are endowed with a cell-intrinsic property termed mitochondrial priming that renders them highly sensitive to apoptotic stimuli. Thus, all attempts to broaden the insights into hESCs apoptosis may be helpful for establishing pro-survival strategies valuable for its in vitro culture and further use in clinical applications. Cyclin-dependent kinases (CDKs), a family of serine/threonine protein kinases originally identified as regulators of the eukaryotic cell cycle, can also regulate transcription and differentiation. Moreover, there are compelling data suggesting that its activities are involved in certain apoptotic programs in different cell types. Currently, it is not completely determined whether CDKs regulate apoptotic processes in rapidly proliferating and apoptosisprone hESCs. In this study, to elucidate the effect of CDKs inhibition in hESCs we used Roscovitine (ROSC), a purine analogue that selectively inhibits the activities of these kinases. Results: Inhibition of CDKs by ROSC triggers programmed cell death in hESCs but not in proliferating somatic cells (human fibroblasts). The apoptotic process encompasses caspase-9 and -3 activation followed by PARP cleavage. ROSC treatment also leads to p53 stabilization, which coincides with site-specific phosphorylation at serine 46 and decreased levels of Mdm2. Additionally, we observed a transcriptional induction of p53AIP1, a repression of pro-survival factor Mcl1 and an up-regulation of pro-apoptotic BH3-only proteins NOXA and PUMA. Importantly, we found that the role of CDK2 inhibition appears to be at best accessory as an active CDK2 is not required to ensure hESCs survival. Conclusion: Our experimental data reveal that hESCs, contrary to fibroblasts, exhibit a pronounced sensitivity to ROSC
Fil: Videla Richardson, Guillermo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Furmento, Verónica Alejandra. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina
Fil: Garcia, Carolina Paola. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Morris Hanon, Olivia. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sevlever, Gustavo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Romorini, Leonardo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Scassa, Maria Elida. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
APOPTOSIS
CYCLIN-DEPENDENT KINASES
HUMAN EMBRYONIC STEM CELLS
ROSCOVITINE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/155445
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/155445
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor RoscovitineVidela Richardson, GuillermoFurmento, Verónica AlejandraGarcia, Carolina PaolaMorris Hanon, OliviaSevlever, GustavoRomorini, LeonardoScassa, Maria ElidaAPOPTOSISCYCLIN-DEPENDENT KINASESHUMAN EMBRYONIC STEM CELLSROSCOVITINEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Background: The essentially unlimited expansion potential and the pluripotency of human embryonic stem cells (hESCs) make them attractive for cell-based therapeutic purposes. Although hESCs can indefinitely proliferate in culture, unlike transformed cancer cells, they are endowed with a cell-intrinsic property termed mitochondrial priming that renders them highly sensitive to apoptotic stimuli. Thus, all attempts to broaden the insights into hESCs apoptosis may be helpful for establishing pro-survival strategies valuable for its in vitro culture and further use in clinical applications. Cyclin-dependent kinases (CDKs), a family of serine/threonine protein kinases originally identified as regulators of the eukaryotic cell cycle, can also regulate transcription and differentiation. Moreover, there are compelling data suggesting that its activities are involved in certain apoptotic programs in different cell types. Currently, it is not completely determined whether CDKs regulate apoptotic processes in rapidly proliferating and apoptosisprone hESCs. In this study, to elucidate the effect of CDKs inhibition in hESCs we used Roscovitine (ROSC), a purine analogue that selectively inhibits the activities of these kinases. Results: Inhibition of CDKs by ROSC triggers programmed cell death in hESCs but not in proliferating somatic cells (human fibroblasts). The apoptotic process encompasses caspase-9 and -3 activation followed by PARP cleavage. ROSC treatment also leads to p53 stabilization, which coincides with site-specific phosphorylation at serine 46 and decreased levels of Mdm2. Additionally, we observed a transcriptional induction of p53AIP1, a repression of pro-survival factor Mcl1 and an up-regulation of pro-apoptotic BH3-only proteins NOXA and PUMA. Importantly, we found that the role of CDK2 inhibition appears to be at best accessory as an active CDK2 is not required to ensure hESCs survival. Conclusion: Our experimental data reveal that hESCs, contrary to fibroblasts, exhibit a pronounced sensitivity to ROSCFil: Videla Richardson, Guillermo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Furmento, Verónica Alejandra. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; ArgentinaFil: Garcia, Carolina Paola. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Morris Hanon, Olivia. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sevlever, Gustavo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Romorini, Leonardo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Scassa, Maria Elida. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaBioMed Central2019-08-28info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/155445Videla Richardson, Guillermo; Furmento, Verónica Alejandra; Garcia, Carolina Paola; Morris Hanon, Olivia; Sevlever, Gustavo; et al.; Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine; BioMed Central; Bmc Cell Biology; 20; 40; 28-8-2019; 1-152661-8850CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://bmcmolcellbiol.biomedcentral.com/articles/10.1186/s12860-019-0222-3info:eu-repo/semantics/altIdentifier/doi/10.1186/s12860-019-0222-3info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:52:54Zoai:ri.conicet.gov.ar:11336/155445instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:52:54.307CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine
title Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine
spellingShingle Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine
Videla Richardson, Guillermo
APOPTOSIS
CYCLIN-DEPENDENT KINASES
HUMAN EMBRYONIC STEM CELLS
ROSCOVITINE
title_short Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine
title_full Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine
title_fullStr Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine
title_full_unstemmed Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine
title_sort Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine
dc.creator.none.fl_str_mv Videla Richardson, Guillermo
Furmento, Verónica Alejandra
Garcia, Carolina Paola
Morris Hanon, Olivia
Sevlever, Gustavo
Romorini, Leonardo
Scassa, Maria Elida
author Videla Richardson, Guillermo
author_facet Videla Richardson, Guillermo
Furmento, Verónica Alejandra
Garcia, Carolina Paola
Morris Hanon, Olivia
Sevlever, Gustavo
Romorini, Leonardo
Scassa, Maria Elida
author_role author
author2 Furmento, Verónica Alejandra
Garcia, Carolina Paola
Morris Hanon, Olivia
Sevlever, Gustavo
Romorini, Leonardo
Scassa, Maria Elida
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv APOPTOSIS
CYCLIN-DEPENDENT KINASES
HUMAN EMBRYONIC STEM CELLS
ROSCOVITINE
topic APOPTOSIS
CYCLIN-DEPENDENT KINASES
HUMAN EMBRYONIC STEM CELLS
ROSCOVITINE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Background: The essentially unlimited expansion potential and the pluripotency of human embryonic stem cells (hESCs) make them attractive for cell-based therapeutic purposes. Although hESCs can indefinitely proliferate in culture, unlike transformed cancer cells, they are endowed with a cell-intrinsic property termed mitochondrial priming that renders them highly sensitive to apoptotic stimuli. Thus, all attempts to broaden the insights into hESCs apoptosis may be helpful for establishing pro-survival strategies valuable for its in vitro culture and further use in clinical applications. Cyclin-dependent kinases (CDKs), a family of serine/threonine protein kinases originally identified as regulators of the eukaryotic cell cycle, can also regulate transcription and differentiation. Moreover, there are compelling data suggesting that its activities are involved in certain apoptotic programs in different cell types. Currently, it is not completely determined whether CDKs regulate apoptotic processes in rapidly proliferating and apoptosisprone hESCs. In this study, to elucidate the effect of CDKs inhibition in hESCs we used Roscovitine (ROSC), a purine analogue that selectively inhibits the activities of these kinases. Results: Inhibition of CDKs by ROSC triggers programmed cell death in hESCs but not in proliferating somatic cells (human fibroblasts). The apoptotic process encompasses caspase-9 and -3 activation followed by PARP cleavage. ROSC treatment also leads to p53 stabilization, which coincides with site-specific phosphorylation at serine 46 and decreased levels of Mdm2. Additionally, we observed a transcriptional induction of p53AIP1, a repression of pro-survival factor Mcl1 and an up-regulation of pro-apoptotic BH3-only proteins NOXA and PUMA. Importantly, we found that the role of CDK2 inhibition appears to be at best accessory as an active CDK2 is not required to ensure hESCs survival. Conclusion: Our experimental data reveal that hESCs, contrary to fibroblasts, exhibit a pronounced sensitivity to ROSC
Fil: Videla Richardson, Guillermo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Furmento, Verónica Alejandra. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina
Fil: Garcia, Carolina Paola. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Morris Hanon, Olivia. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sevlever, Gustavo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Romorini, Leonardo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Scassa, Maria Elida. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Laboratorio de Investigaciones en Neurociencias Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Background: The essentially unlimited expansion potential and the pluripotency of human embryonic stem cells (hESCs) make them attractive for cell-based therapeutic purposes. Although hESCs can indefinitely proliferate in culture, unlike transformed cancer cells, they are endowed with a cell-intrinsic property termed mitochondrial priming that renders them highly sensitive to apoptotic stimuli. Thus, all attempts to broaden the insights into hESCs apoptosis may be helpful for establishing pro-survival strategies valuable for its in vitro culture and further use in clinical applications. Cyclin-dependent kinases (CDKs), a family of serine/threonine protein kinases originally identified as regulators of the eukaryotic cell cycle, can also regulate transcription and differentiation. Moreover, there are compelling data suggesting that its activities are involved in certain apoptotic programs in different cell types. Currently, it is not completely determined whether CDKs regulate apoptotic processes in rapidly proliferating and apoptosisprone hESCs. In this study, to elucidate the effect of CDKs inhibition in hESCs we used Roscovitine (ROSC), a purine analogue that selectively inhibits the activities of these kinases. Results: Inhibition of CDKs by ROSC triggers programmed cell death in hESCs but not in proliferating somatic cells (human fibroblasts). The apoptotic process encompasses caspase-9 and -3 activation followed by PARP cleavage. ROSC treatment also leads to p53 stabilization, which coincides with site-specific phosphorylation at serine 46 and decreased levels of Mdm2. Additionally, we observed a transcriptional induction of p53AIP1, a repression of pro-survival factor Mcl1 and an up-regulation of pro-apoptotic BH3-only proteins NOXA and PUMA. Importantly, we found that the role of CDK2 inhibition appears to be at best accessory as an active CDK2 is not required to ensure hESCs survival. Conclusion: Our experimental data reveal that hESCs, contrary to fibroblasts, exhibit a pronounced sensitivity to ROSC
publishDate 2019
dc.date.none.fl_str_mv 2019-08-28
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/155445
Videla Richardson, Guillermo; Furmento, Verónica Alejandra; Garcia, Carolina Paola; Morris Hanon, Olivia; Sevlever, Gustavo; et al.; Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine; BioMed Central; Bmc Cell Biology; 20; 40; 28-8-2019; 1-15
2661-8850
CONICET Digital
CONICET
url http://hdl.handle.net/11336/155445
identifier_str_mv Videla Richardson, Guillermo; Furmento, Verónica Alejandra; Garcia, Carolina Paola; Morris Hanon, Olivia; Sevlever, Gustavo; et al.; Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine; BioMed Central; Bmc Cell Biology; 20; 40; 28-8-2019; 1-15
2661-8850
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://bmcmolcellbiol.biomedcentral.com/articles/10.1186/s12860-019-0222-3
info:eu-repo/semantics/altIdentifier/doi/10.1186/s12860-019-0222-3
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
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eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
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dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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