A study on the Death-Inducer Obliterator (Dido) function in somatic cell reprogramming to pluripotency

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de Lectura: 18-07-2022

Detalles Bibliográficos
Autor: Talavera Gutiérrez, Amaia
Tipo de recurso: tesis doctoral
Fecha de publicación:2022
País:España
Institución:Universidad Autónoma de Madrid
Repositorio:Biblos-e Archivo. Repositorio Institucional de la UAM
Idioma:inglés
OAI Identifier:oai:repositorio.uam.es:10486/704100
Acceso en línea:http://hdl.handle.net/10486/704100
Access Level:acceso abierto
Palabra clave:Células madre
Síndrome mielodisplástico
Biología y Biomedicina / Biología
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spelling A study on the Death-Inducer Obliterator (Dido) function in somatic cell reprogramming to pluripotencyTalavera Gutiérrez, AmaiaCélulas madreSíndrome mielodisplásticoBiología y Biomedicina / BiologíaTesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de Lectura: 18-07-2022Throughout development, embryonic pluripotent stem cells (ESCs) acquire committed differentiated cell identities. By forced expression of four factors (reprogramming factors), it is possible to reinstate pluripotency in somatic cells; the cells obtained are called induced pluripotent stem cells (iPSCs). Differentiation and reprogramming can be understood as opposed processes. The Death Inducer Obliterator (Dido) gene is highly expressed in pluripotent cells. Lack of Dido exons 3 and 4 is associated with the human myelodysplastic/myeloproliferative syndrome (MDS/MPD). Embryonic development is not affected in Dido exons 3 and 4 depleted mice, while lack of Dido exon 16 (DidoE16) is embryonic lethal at the onset of gastrulation (d8.5). DidoE16 depleted ESCs self-renew, but their differentiation is impaired. Here, we study the role of Dido in the reprogramming to pluripotency process. We first developed a Cre-loxP conditional mouse with a floxed DidoE16 (to circumvent the embryonic lethality of the homozygotic DidoE16 deletion) and crossed it with a reprogrammable mouse that bears the reprogramming factors in a doxycycline-inducible cassette. We also generated reprogrammable mouse lacking Dido exons 3 and 4. We found that deletion of Dido exons 3 and 4 in heterozygosis or homozygosis have no impact in primary mouse embryonic fibroblasts (MEFs) reprogramming and generate bona-fide iPSC. However, hetero- and homozygotic deletion of DidoE16 in primary MEFs reduces the reprogramming efficiency. Therefore, DidoE16, and not Dido exons 3 and 4, have a role in the in vitro induction of pluripotency in MEFs. As possible molecular explanations for that reduction, we identified that DidoE16 depleted MEFs express the senescence-associated marker β-galactosidase, accumulate R-loops, present DNA damage and replication stress. Moreover, immortalization with the herpes virus E6 and E7 oncogenes (inactivates of p53 and Rb) of DidoE16 depleted MEFs not only recover the reprogramming efficiency but surpasses that one of the wild type MEFs. These findings, together with previous evidence of DidoE16 being needed during gastrulation and for a normal ESC differentiation, led us to postulate that DidoE16 facilitates cell identity changes, in particular processes that participate in gastrulation and reprogramming: epithelialto- mesenchymal and mesenchymal-to-epithelial transitions. Learning techniques and creating tools useful for studying the role of DIDO in humans was also an aim of this thesis. We generated bona-fide wild type human iPSC (hiPSC) from mesenchymal stem cells and confirmed their capacity to differentiate into myeloid progenitors. We designed the deletion of DIDOE16 with the CRISPR/Cas9 technique and validated it in HeLa cells. This methodological expertise can be applied for the knock-out of DIDO exons 3 and 4 in our wild type hiPSC, and for MDS/MPD disease modeling. Also, DIDOE16 can be deleted in the same pluripotent cells to study in vitro the role the exon in humansMartínez Alonso, CarlosDepartamento de Biología MolecularFacultad de CienciasCSIC. Centro Nacional de Biotecnología (CNB)20222022-07-18doctoral thesishttp://purl.org/coar/resource_type/c_db06NAhttp://purl.org/coar/version/c_be7fb7dd8ff6fe43info:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/10486/704100reponame:Biblos-e Archivo. Repositorio Institucional de la UAMinstname:Universidad Autónoma de MadridInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:repositorio.uam.es:10486/7041002026-06-23T12:46:27Z
dc.title.none.fl_str_mv A study on the Death-Inducer Obliterator (Dido) function in somatic cell reprogramming to pluripotency
title A study on the Death-Inducer Obliterator (Dido) function in somatic cell reprogramming to pluripotency
spellingShingle A study on the Death-Inducer Obliterator (Dido) function in somatic cell reprogramming to pluripotency
Talavera Gutiérrez, Amaia
Células madre
Síndrome mielodisplástico
Biología y Biomedicina / Biología
title_short A study on the Death-Inducer Obliterator (Dido) function in somatic cell reprogramming to pluripotency
title_full A study on the Death-Inducer Obliterator (Dido) function in somatic cell reprogramming to pluripotency
title_fullStr A study on the Death-Inducer Obliterator (Dido) function in somatic cell reprogramming to pluripotency
title_full_unstemmed A study on the Death-Inducer Obliterator (Dido) function in somatic cell reprogramming to pluripotency
title_sort A study on the Death-Inducer Obliterator (Dido) function in somatic cell reprogramming to pluripotency
dc.creator.none.fl_str_mv Talavera Gutiérrez, Amaia
author Talavera Gutiérrez, Amaia
author_facet Talavera Gutiérrez, Amaia
author_role author
dc.contributor.none.fl_str_mv Martínez Alonso, Carlos
Departamento de Biología Molecular
Facultad de Ciencias
CSIC. Centro Nacional de Biotecnología (CNB)
dc.subject.none.fl_str_mv Células madre
Síndrome mielodisplástico
Biología y Biomedicina / Biología
topic Células madre
Síndrome mielodisplástico
Biología y Biomedicina / Biología
description Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de Lectura: 18-07-2022
publishDate 2022
dc.date.none.fl_str_mv 2022
2022-07-18
dc.type.none.fl_str_mv doctoral thesis
http://purl.org/coar/resource_type/c_db06
NA
http://purl.org/coar/version/c_be7fb7dd8ff6fe43
dc.type.openaire.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
dc.identifier.none.fl_str_mv http://hdl.handle.net/10486/704100
url http://hdl.handle.net/10486/704100
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
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
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Biblos-e Archivo. Repositorio Institucional de la UAM
instname:Universidad Autónoma de Madrid
instname_str Universidad Autónoma de Madrid
reponame_str Biblos-e Archivo. Repositorio Institucional de la UAM
collection Biblos-e Archivo. Repositorio Institucional de la UAM
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,300719

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