title: Direct Conversion of Fibroblasts to Megakaryocyte Progenitors.
abstract: Current sources of platelets for transfusion are insufficient and associated with risk of alloimmunization and blood-borne infection. These limitations could be addressed by the generation of autologous megakaryocytes (MKs) derived in vitro from somatic cells with the ability to engraft and differentiate in vivo. Here, we show that overexpression of a defined set of six transcription factors efficiently converts mouse and human fibroblasts into MK-like progenitors. The transdifferentiated cells are CD41+, display polylobulated nuclei, have ploidies higher than 4N, form MK colonies, and give rise to platelets in vitro. Moreover, transplantation of MK-like murine progenitor cells into NSG mice results in successful engraftment and further maturation in vivo. Similar results are obtained using disease-corrected fibroblasts from Fanconi anemia patients. Our results combined demonstrate that functional MK progenitors with clinical potential can be obtained in vitro, circumventing the use of hematopoietic progenitors or pluripotent stem cells.
authors.affiliation: Center of Regenerative Medicine in Barcelona (CMRB), Barcelona Biomedical Research Park, Dr. Aiguader 88, 08003 Barcelona, Spain; Center for Networked Biomedical Research on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), 28029 Madrid, Spain. Electronic address: jpulecio@cmrb.eu.
authors.forename: Julian
authors.initials: J
authors.lastname: Pulecio
authors.affiliation: Center of Regenerative Medicine in Barcelona (CMRB), Barcelona Biomedical Research Park, Dr. Aiguader 88, 08003 Barcelona, Spain.
authors.forename: Oriol
authors.initials: O
authors.lastname: Alejo-Valle
authors.affiliation: Department of Molecular Medicine and Gene Therapy, Lund Stem Cell Centre, Lund University, 22184 Lund, Sweden.
authors.forename: Sandra
authors.initials: S
authors.lastname: Capellera-Garcia
authors.affiliation: Center of Regenerative Medicine in Barcelona (CMRB), Barcelona Biomedical Research Park, Dr. Aiguader 88, 08003 Barcelona, Spain.
authors.forename: Marianna
authors.initials: M
authors.lastname: Vitaloni
authors.affiliation: Division of Hematopoietic Innovative Therapies, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas and Centro de Investigación Biomédica en Red de Enfermedades Raras (CIEMAT/CIBERER-ISCIII), 28040 Madrid, Spain; Advanced Therapies Unit, I.I.S. Fundación Jiménez Díaz (UAM), 28040 Madrid, Spain.
authors.forename: Paula
authors.initials: P
authors.lastname: Rio
authors.affiliation: Center of Regenerative Medicine in Barcelona (CMRB), Barcelona Biomedical Research Park, Dr. Aiguader 88, 08003 Barcelona, Spain.
authors.forename: Eva
authors.initials: E
authors.lastname: Mejía-Ramírez
authors.affiliation: Center of Regenerative Medicine in Barcelona (CMRB), Barcelona Biomedical Research Park, Dr. Aiguader 88, 08003 Barcelona, Spain.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Caserta
authors.affiliation: Division of Hematopoietic Innovative Therapies, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas and Centro de Investigación Biomédica en Red de Enfermedades Raras (CIEMAT/CIBERER-ISCIII), 28040 Madrid, Spain; Advanced Therapies Unit, I.I.S. Fundación Jiménez Díaz (UAM), 28040 Madrid, Spain.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Bueren
authors.affiliation: Department of Molecular Medicine and Gene Therapy, Lund Stem Cell Centre, Lund University, 22184 Lund, Sweden.
authors.forename: Johan
authors.initials: J
authors.lastname: Flygare
authors.affiliation: Center of Regenerative Medicine in Barcelona (CMRB), Barcelona Biomedical Research Park, Dr. Aiguader 88, 08003 Barcelona, Spain; Center for Networked Biomedical Research on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), 28029 Madrid, Spain; Institució Catalana de Recerca i Estudis Avançats (ICREA), 08010 Barcelona, Spain. Electronic address: araya@cmrb.eu.
authors.forename: Angel
authors.initials: A
authors.lastname: Raya
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D054338:Cell Transdifferentiation; D050676:Core Binding Factor Alpha 2 Subunit / Q000378:metabolism; D005199:Fanconi Anemia / Q000473:pathology; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D050989:GATA2 Transcription Factor / Q000378:metabolism; D006801:Humans; D055016:Megakaryocyte Progenitor Cells / Q000166:cytology* / Q000637:transplantation; D051379:Mice
chemical_list: D050676:Core Binding Factor Alpha 2 Subunit; D050989:GATA2 Transcription Factor
keywords: Fanconi anemia; lineage conversion; platelets; thrombocytopenia; transdifferentiation