14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells

Autores
Rivera, Lautaro; Bustos, Diego Martin; Uhart, Marina
Año de publicación
2021
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
14-3-3proteins constitute a family of regulatory molecules that participatein a plethora of cellular processes mainly through protein-proteininteractions. Even though 14-3-3 protein family members show somefunctional redundancy, there is growing evidence that indicatesevolutionary and biochemical diversity. Consistent with theliterature, previous research from our laboratory showed thatexpression levels of 14-3-3 paralogs are independently regulatedduring the adipogenesis and osteogenesis of human adiposederived-mesenchymal stem cells (hASCs). In the current work, we useda validated approach to isolate hASCs and studied the implication of14-3-3γon the osteogenic commitment of these cells. To address this purpose,we delivered a 14-3-3γshRNA construct into hASCs by pAd-BLOCKiT, an adenoviral vectorcontaining a human U6 promoter, and examined the effect on thedifferentiation potential into osteoblasts. The latter was evaluatedby: i) measuring alkaline phosphatase (ALP) activity, an early-stageosteoblast differentiation biomarker, and ii) detectingRunt-related transcription factor 2 (Runx2, master regulator of boneformation) protein levels. Cells were either maintained for 14 dayswith standard growth media (control, low glucose DMEM; 5% FBS) orinduced with an osteogenic differentiation medium (ODM; an optimizeddrug cocktail that includes dexamethasone, β-glycerophosphate,and 2-phospho-L-ascorbic acid). Our results clearly showed a decreasein both Runx2 protein levels and ALP activity in 14-3-3γdepleted hASCs. This also accords with our earlier observations,which showed that reduced expression of 14-3-3γhad a negative impact on the osteoblastic transdifferentiation ofNIH3T3-L1 cells. Taken together, these findings suggest a regulatoryrole for 14-3-3γin hASC differentiation to the osteogenic lineage.
Fil: Rivera, Lautaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Bustos, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Uhart, Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
LVII Reunión Anual de la Sociedad Argentina de Investigaciónes en Bioquímica y Biología Molecular
Mendoza
Argentina
Sociedad Argentina de Investigaciónes en Bioquímica y Biología Molecular
Materia
14-3-3
osteogenesis
celulas madre
RunX2
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/202254
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/202254
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling 14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cellsRivera, LautaroBustos, Diego MartinUhart, Marina14-3-3osteogenesiscelulas madreRunX2https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/114-3-3proteins constitute a family of regulatory molecules that participatein a plethora of cellular processes mainly through protein-proteininteractions. Even though 14-3-3 protein family members show somefunctional redundancy, there is growing evidence that indicatesevolutionary and biochemical diversity. Consistent with theliterature, previous research from our laboratory showed thatexpression levels of 14-3-3 paralogs are independently regulatedduring the adipogenesis and osteogenesis of human adiposederived-mesenchymal stem cells (hASCs). In the current work, we useda validated approach to isolate hASCs and studied the implication of14-3-3γon the osteogenic commitment of these cells. To address this purpose,we delivered a 14-3-3γshRNA construct into hASCs by pAd-BLOCKiT, an adenoviral vectorcontaining a human U6 promoter, and examined the effect on thedifferentiation potential into osteoblasts. The latter was evaluatedby: i) measuring alkaline phosphatase (ALP) activity, an early-stageosteoblast differentiation biomarker, and ii) detectingRunt-related transcription factor 2 (Runx2, master regulator of boneformation) protein levels. Cells were either maintained for 14 dayswith standard growth media (control, low glucose DMEM; 5% FBS) orinduced with an osteogenic differentiation medium (ODM; an optimizeddrug cocktail that includes dexamethasone, β-glycerophosphate,and 2-phospho-L-ascorbic acid). Our results clearly showed a decreasein both Runx2 protein levels and ALP activity in 14-3-3γdepleted hASCs. This also accords with our earlier observations,which showed that reduced expression of 14-3-3γhad a negative impact on the osteoblastic transdifferentiation ofNIH3T3-L1 cells. Taken together, these findings suggest a regulatoryrole for 14-3-3γin hASC differentiation to the osteogenic lineage.Fil: Rivera, Lautaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Bustos, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Uhart, Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaLVII Reunión Anual de la Sociedad Argentina de Investigaciónes en Bioquímica y Biología MolecularMendozaArgentinaSociedad Argentina de Investigaciónes en Bioquímica y Biología MolecularTech Science Press2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/vnd.openxmlformats-officedocument.wordprocessingml.documentapplication/pdfhttp://hdl.handle.net/11336/20225414-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells; LVII Reunión Anual de la Sociedad Argentina de Investigaciónes en Bioquímica y Biología Molecular; Mendoza; Argentina; 2021; 1-11667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.samige.org.ar/admin/news/files/177-Biocell-Preprint-SAIB-SAMIGE-2021.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:28:18Zoai:ri.conicet.gov.ar:11336/202254instacron: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:34982025-10-22 11:28:18.657CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv 14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells
title 14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells
spellingShingle 14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells
Rivera, Lautaro
14-3-3
osteogenesis
celulas madre
RunX2
title_short 14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells
title_full 14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells
title_fullStr 14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells
title_full_unstemmed 14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells
title_sort 14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells
dc.creator.none.fl_str_mv Rivera, Lautaro
Bustos, Diego Martin
Uhart, Marina
author Rivera, Lautaro
author_facet Rivera, Lautaro
Bustos, Diego Martin
Uhart, Marina
author_role author
author2 Bustos, Diego Martin
Uhart, Marina
author2_role author
author
dc.subject.none.fl_str_mv 14-3-3
osteogenesis
celulas madre
RunX2
topic 14-3-3
osteogenesis
celulas madre
RunX2
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv 14-3-3proteins constitute a family of regulatory molecules that participatein a plethora of cellular processes mainly through protein-proteininteractions. Even though 14-3-3 protein family members show somefunctional redundancy, there is growing evidence that indicatesevolutionary and biochemical diversity. Consistent with theliterature, previous research from our laboratory showed thatexpression levels of 14-3-3 paralogs are independently regulatedduring the adipogenesis and osteogenesis of human adiposederived-mesenchymal stem cells (hASCs). In the current work, we useda validated approach to isolate hASCs and studied the implication of14-3-3γon the osteogenic commitment of these cells. To address this purpose,we delivered a 14-3-3γshRNA construct into hASCs by pAd-BLOCKiT, an adenoviral vectorcontaining a human U6 promoter, and examined the effect on thedifferentiation potential into osteoblasts. The latter was evaluatedby: i) measuring alkaline phosphatase (ALP) activity, an early-stageosteoblast differentiation biomarker, and ii) detectingRunt-related transcription factor 2 (Runx2, master regulator of boneformation) protein levels. Cells were either maintained for 14 dayswith standard growth media (control, low glucose DMEM; 5% FBS) orinduced with an osteogenic differentiation medium (ODM; an optimizeddrug cocktail that includes dexamethasone, β-glycerophosphate,and 2-phospho-L-ascorbic acid). Our results clearly showed a decreasein both Runx2 protein levels and ALP activity in 14-3-3γdepleted hASCs. This also accords with our earlier observations,which showed that reduced expression of 14-3-3γhad a negative impact on the osteoblastic transdifferentiation ofNIH3T3-L1 cells. Taken together, these findings suggest a regulatoryrole for 14-3-3γin hASC differentiation to the osteogenic lineage.
Fil: Rivera, Lautaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Bustos, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Uhart, Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
LVII Reunión Anual de la Sociedad Argentina de Investigaciónes en Bioquímica y Biología Molecular
Mendoza
Argentina
Sociedad Argentina de Investigaciónes en Bioquímica y Biología Molecular
description 14-3-3proteins constitute a family of regulatory molecules that participatein a plethora of cellular processes mainly through protein-proteininteractions. Even though 14-3-3 protein family members show somefunctional redundancy, there is growing evidence that indicatesevolutionary and biochemical diversity. Consistent with theliterature, previous research from our laboratory showed thatexpression levels of 14-3-3 paralogs are independently regulatedduring the adipogenesis and osteogenesis of human adiposederived-mesenchymal stem cells (hASCs). In the current work, we useda validated approach to isolate hASCs and studied the implication of14-3-3γon the osteogenic commitment of these cells. To address this purpose,we delivered a 14-3-3γshRNA construct into hASCs by pAd-BLOCKiT, an adenoviral vectorcontaining a human U6 promoter, and examined the effect on thedifferentiation potential into osteoblasts. The latter was evaluatedby: i) measuring alkaline phosphatase (ALP) activity, an early-stageosteoblast differentiation biomarker, and ii) detectingRunt-related transcription factor 2 (Runx2, master regulator of boneformation) protein levels. Cells were either maintained for 14 dayswith standard growth media (control, low glucose DMEM; 5% FBS) orinduced with an osteogenic differentiation medium (ODM; an optimizeddrug cocktail that includes dexamethasone, β-glycerophosphate,and 2-phospho-L-ascorbic acid). Our results clearly showed a decreasein both Runx2 protein levels and ALP activity in 14-3-3γdepleted hASCs. This also accords with our earlier observations,which showed that reduced expression of 14-3-3γhad a negative impact on the osteoblastic transdifferentiation ofNIH3T3-L1 cells. Taken together, these findings suggest a regulatoryrole for 14-3-3γin hASC differentiation to the osteogenic lineage.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/202254
14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells; LVII Reunión Anual de la Sociedad Argentina de Investigaciónes en Bioquímica y Biología Molecular; Mendoza; Argentina; 2021; 1-1
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/202254
identifier_str_mv 14-3-3γ silencing impairs osteogenic differentiation of human adipose derived-mesenchymal stem cells; LVII Reunión Anual de la Sociedad Argentina de Investigaciónes en Bioquímica y Biología Molecular; Mendoza; Argentina; 2021; 1-1
1667-5746
CONICET Digital
CONICET
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language eng
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eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
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application/vnd.openxmlformats-officedocument.wordprocessingml.document
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dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Tech Science Press
publisher.none.fl_str_mv Tech Science Press
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