A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.

Alvarez-Palomo, Ana Belén; Requena-Osete, Jordi; Delgado-Morales, Raul; Moreno-Manzano, Victoria; Grau-Bove, Carme; Tejera, Agueda M; Otero, Manel Juan; Barrot, Carme; Santos-Barriopedro, Irene; Vaquero, Alejandro; Mezquita-Pla, Jovita; Moran, Sebastian; Naya, Carlos Hobeich; Garcia-Martínez, Iris; Pérez, Francisco Vidal; Blasco, MA; Esteller, Manel; Edel, Michael J

A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.

A key challenge for clinical application of induced pluripotent stem cells (iPSC) to accurately model and treat human pathologies depends on developing a method to generate genetically stable cells to reduce long-term risks of cell transplant therapy. Here, we hypothesized that CYCLIN D1 repairs DNA...

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Autores:	Alvarez-Palomo, Ana Belén, Requena-Osete, Jordi, Delgado-Morales, Raul, Moreno-Manzano, Victoria, Grau-Bove, Carme, Tejera, Agueda M, Otero, Manel Juan, Barrot, Carme, Santos-Barriopedro, Irene, Vaquero, Alejandro, Mezquita-Pla, Jovita, Moran, Sebastian, Naya, Carlos Hobeich, Garcia-Martínez, Iris, Pérez, Francisco Vidal, Blasco, MA, Esteller, Manel, Edel, Michael J
Formato:	artículo
Fecha de publicación:	2021
País:	España
Recursos:	Instituto de Salud Carlos III (ISCIII)
Repositorio:	Repisalud
Idioma:	inglés
OAI Identifier:	oai:repisalud.isciii.es:20.500.12105/19030
Acesso em linha:	http://hdl.handle.net/20.500.12105/19030
Access Level:	acceso abierto
Palavra-chave:	Induced Pluripotent Stem Cells Animals Cell Differentiation Cellular Reprogramming Cyclin D1 DNA Repair Humans Mice RNA, Messenger

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spelling	A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.Alvarez-Palomo, Ana BelénRequena-Osete, JordiDelgado-Morales, RaulMoreno-Manzano, VictoriaGrau-Bove, CarmeTejera, Agueda MOtero, Manel JuanBarrot, CarmeSantos-Barriopedro, IreneVaquero, AlejandroMezquita-Pla, JovitaMoran, SebastianNaya, Carlos HobeichGarcia-Martínez, IrisPérez, Francisco VidalBlasco, MAEsteller, ManelEdel, Michael JInduced Pluripotent Stem CellsAnimalsCell DifferentiationCellular ReprogrammingCyclin D1DNA RepairHumansMiceRNA, MessengerA key challenge for clinical application of induced pluripotent stem cells (iPSC) to accurately model and treat human pathologies depends on developing a method to generate genetically stable cells to reduce long-term risks of cell transplant therapy. Here, we hypothesized that CYCLIN D1 repairs DNA by highly efficient homologous recombination (HR) during reprogramming to iPSC that reduces genetic instability and threat of neoplastic growth. We adopted a synthetic mRNA transfection method using clinically compatible conditions with CYCLIN D1 plus base factors (OCT3/4, SOX2, KLF4, LIN28) and compared with methods that use C-MYC. We demonstrate that CYCLIN D1 made iPSC have (a) lower multitelomeric signal, (b) reduced double-strand DNA breaks, (c) correct nuclear localization of RAD51 protein expression, and (d) reduced single-nucleotide polymorphism (SNP) changes per chromosome, compared with the classical reprogramming method using C-MYC. CYCLIN D1 iPSC have reduced teratoma Ki67 cell growth kinetics and derived neural stem cells successfully engraft in a hostile spinal cord injury (SCI) microenvironment with efficient survival, differentiation. We demonstrate that CYCLIN D1 promotes double-stranded DNA damage repair predominantly through HR during cell reprogramming to efficiently produce iPSC. CYCLIN D1 reduces general cell stress associated with significantly lower SIRT1 gene expression and can rescue Sirt1 null mouse cell reprogramming. In conclusion, we show synthetic mRNA transfection of CYCLIN D1 repairs DNA during reprogramming resulting in significantly improved genetically stable footprint in human iPSC, enabling a new cell reprogramming method for more accurate and reliable generation of human iPSC for disease modeling and future clinical applications.Oxford University PressFundación La Marató TV3Government of Catalonia (España)Worldwide Cancer ResearchFundación Banco SantanderInstituto de Salud Carlos IIIUniversity of Barcelona (España)Agencia Estatal de Investigación (España)Ministerio de Economía e Innovación (España)20242024-03-2120212021-07-0120212021-07-01journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/20.500.12105/19030reponame:Repisaludinstname:Instituto de Salud Carlos III (ISCIII)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution-NonCommercial-NoDerivatives 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:repisalud.isciii.es:20.500.12105/190302026-06-12T12:43:37Z
dc.title.none.fl_str_mv	A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.
title	A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.
spellingShingle	A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells. Alvarez-Palomo, Ana Belén Induced Pluripotent Stem Cells Animals Cell Differentiation Cellular Reprogramming Cyclin D1 DNA Repair Humans Mice RNA, Messenger
title_short	A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.
title_full	A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.
title_fullStr	A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.
title_full_unstemmed	A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.
title_sort	A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.
dc.creator.none.fl_str_mv	Alvarez-Palomo, Ana Belén Requena-Osete, Jordi Delgado-Morales, Raul Moreno-Manzano, Victoria Grau-Bove, Carme Tejera, Agueda M Otero, Manel Juan Barrot, Carme Santos-Barriopedro, Irene Vaquero, Alejandro Mezquita-Pla, Jovita Moran, Sebastian Naya, Carlos Hobeich Garcia-Martínez, Iris Pérez, Francisco Vidal Blasco, MA Esteller, Manel Edel, Michael J
author	Alvarez-Palomo, Ana Belén
author_facet	Alvarez-Palomo, Ana Belén Requena-Osete, Jordi Delgado-Morales, Raul Moreno-Manzano, Victoria Grau-Bove, Carme Tejera, Agueda M Otero, Manel Juan Barrot, Carme Santos-Barriopedro, Irene Vaquero, Alejandro Mezquita-Pla, Jovita Moran, Sebastian Naya, Carlos Hobeich Garcia-Martínez, Iris Pérez, Francisco Vidal Blasco, MA Esteller, Manel Edel, Michael J
author_role	author
author2	Requena-Osete, Jordi Delgado-Morales, Raul Moreno-Manzano, Victoria Grau-Bove, Carme Tejera, Agueda M Otero, Manel Juan Barrot, Carme Santos-Barriopedro, Irene Vaquero, Alejandro Mezquita-Pla, Jovita Moran, Sebastian Naya, Carlos Hobeich Garcia-Martínez, Iris Pérez, Francisco Vidal Blasco, MA Esteller, Manel Edel, Michael J
author2_role	author author author author author author author author author author author author author author author author author
dc.contributor.none.fl_str_mv	Fundación La Marató TV3 Government of Catalonia (España) Worldwide Cancer Research Fundación Banco Santander Instituto de Salud Carlos III University of Barcelona (España) Agencia Estatal de Investigación (España) Ministerio de Economía e Innovación (España)
dc.subject.none.fl_str_mv	Induced Pluripotent Stem Cells Animals Cell Differentiation Cellular Reprogramming Cyclin D1 DNA Repair Humans Mice RNA, Messenger
topic	Induced Pluripotent Stem Cells Animals Cell Differentiation Cellular Reprogramming Cyclin D1 DNA Repair Humans Mice RNA, Messenger
description	A key challenge for clinical application of induced pluripotent stem cells (iPSC) to accurately model and treat human pathologies depends on developing a method to generate genetically stable cells to reduce long-term risks of cell transplant therapy. Here, we hypothesized that CYCLIN D1 repairs DNA by highly efficient homologous recombination (HR) during reprogramming to iPSC that reduces genetic instability and threat of neoplastic growth. We adopted a synthetic mRNA transfection method using clinically compatible conditions with CYCLIN D1 plus base factors (OCT3/4, SOX2, KLF4, LIN28) and compared with methods that use C-MYC. We demonstrate that CYCLIN D1 made iPSC have (a) lower multitelomeric signal, (b) reduced double-strand DNA breaks, (c) correct nuclear localization of RAD51 protein expression, and (d) reduced single-nucleotide polymorphism (SNP) changes per chromosome, compared with the classical reprogramming method using C-MYC. CYCLIN D1 iPSC have reduced teratoma Ki67 cell growth kinetics and derived neural stem cells successfully engraft in a hostile spinal cord injury (SCI) microenvironment with efficient survival, differentiation. We demonstrate that CYCLIN D1 promotes double-stranded DNA damage repair predominantly through HR during cell reprogramming to efficiently produce iPSC. CYCLIN D1 reduces general cell stress associated with significantly lower SIRT1 gene expression and can rescue Sirt1 null mouse cell reprogramming. In conclusion, we show synthetic mRNA transfection of CYCLIN D1 repairs DNA during reprogramming resulting in significantly improved genetically stable footprint in human iPSC, enabling a new cell reprogramming method for more accurate and reliable generation of human iPSC for disease modeling and future clinical applications.
publishDate	2021
dc.date.none.fl_str_mv	2021 2021-07-01 2021 2021-07-01 2024 2024-03-21
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/20.500.12105/19030
url	http://hdl.handle.net/20.500.12105/19030
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 Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/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 Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv	openAccess
dc.format.none.fl_str_mv	application/pdf
dc.publisher.none.fl_str_mv	Oxford University Press
publisher.none.fl_str_mv	Oxford University Press
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
collection	Repisalud
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score	15,812455

A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair, generating improved genetically stable human induced pluripotent stem cells.

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