Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line

Autores
Converti, Ayelén; Bianchi, Maria Silvia; Montaner, Alejandro Daniel; Libertun, Carlos; Lux, Victoria Adela R.; Bonaventura, Maria Marta
Año de publicación
2019
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
We have previously demonstrated that treatment with IMT504 promotes significant improvement in the diabetic condition in diverse animal models. We have also shown effects on gene expression on freshly isolated islets from diabetic IMT504-treated animals. Based on these results, here we evaluated if the effects of IMT504 observed in vivo were due to direct effects on beta cells. In particular we studied cell viability, enzyme activation and gene expression. A murine beta cell line (MIN6B1) was used. Cells were cultured in DMEM with 20 mM glucose, 15% SFB, 71 uM βmercaptoethanol. Cell viability was analized by MTS: cells were stimulated for 24 or 48 h with 0 (C), 2 (IMT2), 4 (IMT4) and 8 ug/ml (IMT8) of IMT504 in DMEM, 20 mM glucose, 2% SFB, 71 uM βmercaptoethanol. Gene expression of Pdx1, Ins2, Ins1 and Mafa was analized by qPCR, using cyclophilin as housekeeping gene. Phosphorylation of proteins of interest was analyzed by Western Blot. Cells were stimulated for 24 and 48 h with IMT504 in DMEM, 20 mM glucose, 0.5% BSA, 71 uM βmercaptoethanol. Enzyme phosphorylaton was also assayed at short time stimulation periods i.e. 0, 5, 15 ,30 and 60 min. For gene expression the dosis of IMT504 used were 0 (C), 0.4 (IMT0.4), 2.2 (IMT2.2) and 4 (IMT4) ug/ml; and for Western Blot were 0 (C), 2 (IMT2), 4 (IMT) and 8 (IMT8) ug/ml. No differences in cell viability were observed at the time points studied (ANOVA for repeated measures: NS). Expression of Pdx1 and Ins2 was significantly increased by 48 h stimulation with IMT504 [ANOVA for repeated measures: Pdx1 (A.U.): C=0.93±0.05; IMT0.4=0.80±0.06; IMT2.2=1.04±0.09; IMT4=1.37±0.09 p<0.05, IMT4 different from C and IMT0.4; Ins2 (A.U.): C=0.99±0.04; IMT0.4=1.04±0.06; IMT2.2=1.13±0.06; IMT4=1.52±0.08 p<0.05, IMT4 different from C and IMT0.4], while no changes in Ins1 and Mafa were observed (NS). pGSK3β/GSK3β ratio (A.U.) and pAkt/Akt ratio (A.U.) were calculated and informed as fold increase with regard to 0 min. At 60 min, pGSK3β/GSK3β ratio was increased compared to 0 min (ANOVA with repeated measures: pGSK3β/GSK3β ratio (A.U.) 0 min=1, 5 min=1.15±0.06, 15 min=1.20±0.05; 30 min=1.54±0.14; 60 min=2.04±0.17, 60 min different from 0 min, p<0.03). No significant differences were observed in pAkt/Akt ratio (ANOVA with repeated measures, NS). For 24 and 48 hs we found no significant differences between groups. Our results demonstrate a direct effect of IMT504 on gene expression in beta cells and suggest that IMT504 could exert its actions on beta cells through a pathway that includes GSK3β phosphorylation. Further studies must be done to dilucidate their implications on beta cell function recovery in diabetic animals.
Fil: Converti, Ayelén. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Bianchi, Maria Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Montaner, Alejandro Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ciencia y Tecnología "Dr. César Milstein". Fundación Pablo Cassará. Instituto de Ciencia y Tecnología "Dr. César Milstein"; Argentina
Fil: Libertun, Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Lux, Victoria Adela R.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Bonaventura, Maria Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
ENDO 2019: 101st Annual Meeting of the Endocrine Society
New Orleans
Estados Unidos
The Endocrine Society
Materia
IMT504
MIN6-B1 CELLS
GENE EXPRESSION
GSK3 PHOSPHORYLATION
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/244044
Acceder
id CONICETDig_244ed1e8a141676c2542175c8eba5b58
oai_identifier_str oai:ri.conicet.gov.ar:11336/244044
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell lineConverti, AyelénBianchi, Maria SilviaMontaner, Alejandro DanielLibertun, CarlosLux, Victoria Adela R.Bonaventura, Maria MartaIMT504MIN6-B1 CELLSGENE EXPRESSIONGSK3 PHOSPHORYLATIONhttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3We have previously demonstrated that treatment with IMT504 promotes significant improvement in the diabetic condition in diverse animal models. We have also shown effects on gene expression on freshly isolated islets from diabetic IMT504-treated animals. Based on these results, here we evaluated if the effects of IMT504 observed in vivo were due to direct effects on beta cells. In particular we studied cell viability, enzyme activation and gene expression. A murine beta cell line (MIN6B1) was used. Cells were cultured in DMEM with 20 mM glucose, 15% SFB, 71 uM βmercaptoethanol. Cell viability was analized by MTS: cells were stimulated for 24 or 48 h with 0 (C), 2 (IMT2), 4 (IMT4) and 8 ug/ml (IMT8) of IMT504 in DMEM, 20 mM glucose, 2% SFB, 71 uM βmercaptoethanol. Gene expression of Pdx1, Ins2, Ins1 and Mafa was analized by qPCR, using cyclophilin as housekeeping gene. Phosphorylation of proteins of interest was analyzed by Western Blot. Cells were stimulated for 24 and 48 h with IMT504 in DMEM, 20 mM glucose, 0.5% BSA, 71 uM βmercaptoethanol. Enzyme phosphorylaton was also assayed at short time stimulation periods i.e. 0, 5, 15 ,30 and 60 min. For gene expression the dosis of IMT504 used were 0 (C), 0.4 (IMT0.4), 2.2 (IMT2.2) and 4 (IMT4) ug/ml; and for Western Blot were 0 (C), 2 (IMT2), 4 (IMT) and 8 (IMT8) ug/ml. No differences in cell viability were observed at the time points studied (ANOVA for repeated measures: NS). Expression of Pdx1 and Ins2 was significantly increased by 48 h stimulation with IMT504 [ANOVA for repeated measures: Pdx1 (A.U.): C=0.93±0.05; IMT0.4=0.80±0.06; IMT2.2=1.04±0.09; IMT4=1.37±0.09 p<0.05, IMT4 different from C and IMT0.4; Ins2 (A.U.): C=0.99±0.04; IMT0.4=1.04±0.06; IMT2.2=1.13±0.06; IMT4=1.52±0.08 p<0.05, IMT4 different from C and IMT0.4], while no changes in Ins1 and Mafa were observed (NS). pGSK3β/GSK3β ratio (A.U.) and pAkt/Akt ratio (A.U.) were calculated and informed as fold increase with regard to 0 min. At 60 min, pGSK3β/GSK3β ratio was increased compared to 0 min (ANOVA with repeated measures: pGSK3β/GSK3β ratio (A.U.) 0 min=1, 5 min=1.15±0.06, 15 min=1.20±0.05; 30 min=1.54±0.14; 60 min=2.04±0.17, 60 min different from 0 min, p<0.03). No significant differences were observed in pAkt/Akt ratio (ANOVA with repeated measures, NS). For 24 and 48 hs we found no significant differences between groups. Our results demonstrate a direct effect of IMT504 on gene expression in beta cells and suggest that IMT504 could exert its actions on beta cells through a pathway that includes GSK3β phosphorylation. Further studies must be done to dilucidate their implications on beta cell function recovery in diabetic animals.Fil: Converti, Ayelén. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Bianchi, Maria Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Montaner, Alejandro Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ciencia y Tecnología "Dr. César Milstein". Fundación Pablo Cassará. Instituto de Ciencia y Tecnología "Dr. César Milstein"; ArgentinaFil: Libertun, Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Lux, Victoria Adela R.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Bonaventura, Maria Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaENDO 2019: 101st Annual Meeting of the Endocrine SocietyNew OrleansEstados UnidosThe Endocrine SocietyOxford University Press2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectEncuentroJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/244044Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line; ENDO 2019: 101st Annual Meeting of the Endocrine Society; New Orleans; Estados Unidos; 2019; 1-22472-1972CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1210/js.2019-SAT-165info:eu-repo/semantics/altIdentifier/url/https://academic.oup.com/jes/article/3/Supplement_1/SAT-165/5483417Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:50:36Zoai:ri.conicet.gov.ar:11336/244044instacron: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:34982025-09-03 09:50:36.981CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line
title Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line
spellingShingle Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line
Converti, Ayelén
IMT504
MIN6-B1 CELLS
GENE EXPRESSION
GSK3 PHOSPHORYLATION
title_short Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line
title_full Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line
title_fullStr Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line
title_full_unstemmed Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line
title_sort Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line
dc.creator.none.fl_str_mv Converti, Ayelén
Bianchi, Maria Silvia
Montaner, Alejandro Daniel
Libertun, Carlos
Lux, Victoria Adela R.
Bonaventura, Maria Marta
author Converti, Ayelén
author_facet Converti, Ayelén
Bianchi, Maria Silvia
Montaner, Alejandro Daniel
Libertun, Carlos
Lux, Victoria Adela R.
Bonaventura, Maria Marta
author_role author
author2 Bianchi, Maria Silvia
Montaner, Alejandro Daniel
Libertun, Carlos
Lux, Victoria Adela R.
Bonaventura, Maria Marta
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv IMT504
MIN6-B1 CELLS
GENE EXPRESSION
GSK3 PHOSPHORYLATION
topic IMT504
MIN6-B1 CELLS
GENE EXPRESSION
GSK3 PHOSPHORYLATION
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv We have previously demonstrated that treatment with IMT504 promotes significant improvement in the diabetic condition in diverse animal models. We have also shown effects on gene expression on freshly isolated islets from diabetic IMT504-treated animals. Based on these results, here we evaluated if the effects of IMT504 observed in vivo were due to direct effects on beta cells. In particular we studied cell viability, enzyme activation and gene expression. A murine beta cell line (MIN6B1) was used. Cells were cultured in DMEM with 20 mM glucose, 15% SFB, 71 uM βmercaptoethanol. Cell viability was analized by MTS: cells were stimulated for 24 or 48 h with 0 (C), 2 (IMT2), 4 (IMT4) and 8 ug/ml (IMT8) of IMT504 in DMEM, 20 mM glucose, 2% SFB, 71 uM βmercaptoethanol. Gene expression of Pdx1, Ins2, Ins1 and Mafa was analized by qPCR, using cyclophilin as housekeeping gene. Phosphorylation of proteins of interest was analyzed by Western Blot. Cells were stimulated for 24 and 48 h with IMT504 in DMEM, 20 mM glucose, 0.5% BSA, 71 uM βmercaptoethanol. Enzyme phosphorylaton was also assayed at short time stimulation periods i.e. 0, 5, 15 ,30 and 60 min. For gene expression the dosis of IMT504 used were 0 (C), 0.4 (IMT0.4), 2.2 (IMT2.2) and 4 (IMT4) ug/ml; and for Western Blot were 0 (C), 2 (IMT2), 4 (IMT) and 8 (IMT8) ug/ml. No differences in cell viability were observed at the time points studied (ANOVA for repeated measures: NS). Expression of Pdx1 and Ins2 was significantly increased by 48 h stimulation with IMT504 [ANOVA for repeated measures: Pdx1 (A.U.): C=0.93±0.05; IMT0.4=0.80±0.06; IMT2.2=1.04±0.09; IMT4=1.37±0.09 p<0.05, IMT4 different from C and IMT0.4; Ins2 (A.U.): C=0.99±0.04; IMT0.4=1.04±0.06; IMT2.2=1.13±0.06; IMT4=1.52±0.08 p<0.05, IMT4 different from C and IMT0.4], while no changes in Ins1 and Mafa were observed (NS). pGSK3β/GSK3β ratio (A.U.) and pAkt/Akt ratio (A.U.) were calculated and informed as fold increase with regard to 0 min. At 60 min, pGSK3β/GSK3β ratio was increased compared to 0 min (ANOVA with repeated measures: pGSK3β/GSK3β ratio (A.U.) 0 min=1, 5 min=1.15±0.06, 15 min=1.20±0.05; 30 min=1.54±0.14; 60 min=2.04±0.17, 60 min different from 0 min, p<0.03). No significant differences were observed in pAkt/Akt ratio (ANOVA with repeated measures, NS). For 24 and 48 hs we found no significant differences between groups. Our results demonstrate a direct effect of IMT504 on gene expression in beta cells and suggest that IMT504 could exert its actions on beta cells through a pathway that includes GSK3β phosphorylation. Further studies must be done to dilucidate their implications on beta cell function recovery in diabetic animals.
Fil: Converti, Ayelén. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Bianchi, Maria Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Montaner, Alejandro Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ciencia y Tecnología "Dr. César Milstein". Fundación Pablo Cassará. Instituto de Ciencia y Tecnología "Dr. César Milstein"; Argentina
Fil: Libertun, Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Lux, Victoria Adela R.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Bonaventura, Maria Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
ENDO 2019: 101st Annual Meeting of the Endocrine Society
New Orleans
Estados Unidos
The Endocrine Society
description We have previously demonstrated that treatment with IMT504 promotes significant improvement in the diabetic condition in diverse animal models. We have also shown effects on gene expression on freshly isolated islets from diabetic IMT504-treated animals. Based on these results, here we evaluated if the effects of IMT504 observed in vivo were due to direct effects on beta cells. In particular we studied cell viability, enzyme activation and gene expression. A murine beta cell line (MIN6B1) was used. Cells were cultured in DMEM with 20 mM glucose, 15% SFB, 71 uM βmercaptoethanol. Cell viability was analized by MTS: cells were stimulated for 24 or 48 h with 0 (C), 2 (IMT2), 4 (IMT4) and 8 ug/ml (IMT8) of IMT504 in DMEM, 20 mM glucose, 2% SFB, 71 uM βmercaptoethanol. Gene expression of Pdx1, Ins2, Ins1 and Mafa was analized by qPCR, using cyclophilin as housekeeping gene. Phosphorylation of proteins of interest was analyzed by Western Blot. Cells were stimulated for 24 and 48 h with IMT504 in DMEM, 20 mM glucose, 0.5% BSA, 71 uM βmercaptoethanol. Enzyme phosphorylaton was also assayed at short time stimulation periods i.e. 0, 5, 15 ,30 and 60 min. For gene expression the dosis of IMT504 used were 0 (C), 0.4 (IMT0.4), 2.2 (IMT2.2) and 4 (IMT4) ug/ml; and for Western Blot were 0 (C), 2 (IMT2), 4 (IMT) and 8 (IMT8) ug/ml. No differences in cell viability were observed at the time points studied (ANOVA for repeated measures: NS). Expression of Pdx1 and Ins2 was significantly increased by 48 h stimulation with IMT504 [ANOVA for repeated measures: Pdx1 (A.U.): C=0.93±0.05; IMT0.4=0.80±0.06; IMT2.2=1.04±0.09; IMT4=1.37±0.09 p<0.05, IMT4 different from C and IMT0.4; Ins2 (A.U.): C=0.99±0.04; IMT0.4=1.04±0.06; IMT2.2=1.13±0.06; IMT4=1.52±0.08 p<0.05, IMT4 different from C and IMT0.4], while no changes in Ins1 and Mafa were observed (NS). pGSK3β/GSK3β ratio (A.U.) and pAkt/Akt ratio (A.U.) were calculated and informed as fold increase with regard to 0 min. At 60 min, pGSK3β/GSK3β ratio was increased compared to 0 min (ANOVA with repeated measures: pGSK3β/GSK3β ratio (A.U.) 0 min=1, 5 min=1.15±0.06, 15 min=1.20±0.05; 30 min=1.54±0.14; 60 min=2.04±0.17, 60 min different from 0 min, p<0.03). No significant differences were observed in pAkt/Akt ratio (ANOVA with repeated measures, NS). For 24 and 48 hs we found no significant differences between groups. Our results demonstrate a direct effect of IMT504 on gene expression in beta cells and suggest that IMT504 could exert its actions on beta cells through a pathway that includes GSK3β phosphorylation. Further studies must be done to dilucidate their implications on beta cell function recovery in diabetic animals.
publishDate 2019
dc.date.none.fl_str_mv 2019
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Encuentro
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/11336/244044
Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line; ENDO 2019: 101st Annual Meeting of the Endocrine Society; New Orleans; Estados Unidos; 2019; 1-2
2472-1972
CONICET Digital
CONICET
url http://hdl.handle.net/11336/244044
identifier_str_mv Does oligodeoxynucleotide IMT504 have any effect on beta cells? A preliminary study on MIN6B1 cell line; ENDO 2019: 101st Annual Meeting of the Endocrine Society; New Orleans; Estados Unidos; 2019; 1-2
2472-1972
CONICET Digital
CONICET
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dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Oxford University Press
publisher.none.fl_str_mv Oxford University Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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