Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model

The Wiskott-Aldrich syndrome (WAS) is an X-linked primary immunodeficiency caused by mutations in the WAS gene and characterized by severe thrombocytopenia. Although the role of WASp in terminally differentiated lymphocytes and myeloid cells is well characterized, its role in early hematopoietic dif...

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Main Authors:	Toscano, Miguel G, Muñoz, Pilar, Sánchez-Gilabert, Almudena, Cobo, Marién, Benabdellah, Karim, Anderson, Per, Ramos-Mejía, Verónica, Real, Pedro J, Neth, Olaf, Molinos-Quintana, Agueda, Gregory, Philip D, Holmes, Michael C, Martin, Francisco
Format:	Online
Language:	English
Published:	Nature Publishing Group 2016
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4817813/

id	pubmed-4817813
recordtype	oai_dc
spelling	pubmed-48178132016-04-15 Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model Toscano, Miguel G Muñoz, Pilar Sánchez-Gilabert, Almudena Cobo, Marién Benabdellah, Karim Anderson, Per Ramos-Mejía, Verónica Real, Pedro J Neth, Olaf Molinos-Quintana, Agueda Gregory, Philip D Holmes, Michael C Martin, Francisco Original Article The Wiskott-Aldrich syndrome (WAS) is an X-linked primary immunodeficiency caused by mutations in the WAS gene and characterized by severe thrombocytopenia. Although the role of WASp in terminally differentiated lymphocytes and myeloid cells is well characterized, its role in early hematopoietic differentiation and in platelets (Plts) biology is poorly understood. In the present manuscript, we have used zinc finger nucleases targeted to the WAS locus for the development of two isogenic WAS knockout (WASKO) human embryonic stem cell lines (hESCs). Upon hematopoietic differentiation, hESCs-WASKO generated increased ratios of CD34+CD45+ progenitors with altered responses to stem cell factor compared to hESCs-WT. When differentiated toward the megakaryocytic linage, hESCs-WASKO produced increased numbers of CD34+CD41+ progenitors, megakaryocytes (MKs), and Plts. hESCs-WASKO-derived MKs and Plts showed altered phenotype as well as defective responses to agonist, mimicking WAS patients MKs and Plts defects. Interestingly, the defects were more evident in WASp-deficient MKs than in WASp-deficient Plts. Importantly, ectopic WAS expression using lentiviral vectors restored normal Plts development and MKs responses. These data validate the AND-1_WASKO cell lines as a human cellular model for basic research and for preclinical studies for WAS. Nature Publishing Group 2016-02 2015-12-08 /pmc/articles/PMC4817813/ /pubmed/26502776 http://dx.doi.org/10.1038/mt.2015.196 Text en Copyright © 2016 Official journal of the American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Toscano, Miguel G Muñoz, Pilar Sánchez-Gilabert, Almudena Cobo, Marién Benabdellah, Karim Anderson, Per Ramos-Mejía, Verónica Real, Pedro J Neth, Olaf Molinos-Quintana, Agueda Gregory, Philip D Holmes, Michael C Martin, Francisco
spellingShingle	Toscano, Miguel G Muñoz, Pilar Sánchez-Gilabert, Almudena Cobo, Marién Benabdellah, Karim Anderson, Per Ramos-Mejía, Verónica Real, Pedro J Neth, Olaf Molinos-Quintana, Agueda Gregory, Philip D Holmes, Michael C Martin, Francisco Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model
author_facet	Toscano, Miguel G Muñoz, Pilar Sánchez-Gilabert, Almudena Cobo, Marién Benabdellah, Karim Anderson, Per Ramos-Mejía, Verónica Real, Pedro J Neth, Olaf Molinos-Quintana, Agueda Gregory, Philip D Holmes, Michael C Martin, Francisco
author_sort	Toscano, Miguel G
title	Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model
title_short	Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model
title_full	Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model
title_fullStr	Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model
title_full_unstemmed	Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model
title_sort	absence of wasp enhances hematopoietic and megakaryocytic differentiation in a human embryonic stem cell model
description	The Wiskott-Aldrich syndrome (WAS) is an X-linked primary immunodeficiency caused by mutations in the WAS gene and characterized by severe thrombocytopenia. Although the role of WASp in terminally differentiated lymphocytes and myeloid cells is well characterized, its role in early hematopoietic differentiation and in platelets (Plts) biology is poorly understood. In the present manuscript, we have used zinc finger nucleases targeted to the WAS locus for the development of two isogenic WAS knockout (WASKO) human embryonic stem cell lines (hESCs). Upon hematopoietic differentiation, hESCs-WASKO generated increased ratios of CD34+CD45+ progenitors with altered responses to stem cell factor compared to hESCs-WT. When differentiated toward the megakaryocytic linage, hESCs-WASKO produced increased numbers of CD34+CD41+ progenitors, megakaryocytes (MKs), and Plts. hESCs-WASKO-derived MKs and Plts showed altered phenotype as well as defective responses to agonist, mimicking WAS patients MKs and Plts defects. Interestingly, the defects were more evident in WASp-deficient MKs than in WASp-deficient Plts. Importantly, ectopic WAS expression using lentiviral vectors restored normal Plts development and MKs responses. These data validate the AND-1_WASKO cell lines as a human cellular model for basic research and for preclinical studies for WAS.
publisher	Nature Publishing Group
publishDate	2016
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4817813/
_version_	1613561052726820864

Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model

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