Absence of WASp Enhances Hematopoietic and Megakaryocytic Differentiation in a Human Embryonic Stem Cell Model
| Autor: | Miguel G. Toscano, Philip D. Gregory, Olaf Neth, Francisco Martin, Almudena Sánchez-Gilabert, Karim Benabdellah, Marién Cobo, Pilar Muñoz, Per Anderson, Pedro J. Real, Verónica Ramos-Mejía, Michael C. Holmes, Agueda Molinos-Quintana |
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| Rok vydání: | 2016 |
| Předmět: |
Platelet Membrane Glycoprotein IIb
0301 basic medicine Cellular differentiation CD34 Antigens CD34 Stem cell factor Models Biological Cell Line Gene Knockout Techniques 03 medical and health sciences Drug Discovery Genetics Humans Progenitor cell Molecular Biology Embryonic Stem Cells Pharmacology biology Wiskott–Aldrich syndrome protein Cell Differentiation Hematopoietic Stem Cells Embryonic stem cell Cell biology Haematopoiesis 030104 developmental biology Cell culture Immunology biology.protein Leukocyte Common Antigens Molecular Medicine Original Article Megakaryocytes Wiskott-Aldrich Syndrome Protein |
| Zdroj: | Molecular Therapy |
| ISSN: | 1525-0016 |
| DOI: | 10.1038/mt.2015.196 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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