CRISPR-engineered human GATA2 deficiency model uncovers mitotic dysfunction and premature aging in HSPCs, impairing hematopoietic fitness
GATA2 deficiency is a monogenic transcriptopathy disorder characterized by bone marrow failure (BMF), immunodeficiency, and a high risk of developing myelodysplastic neoplasms (MDS) and acute myeloid leukemia (AML). Although informative mouse models have been developed, the mechanisms by which GATA2 haploinsufficiency drives disease initiation in humans remain incompletely understood. To address this, we developed a novel humanized model using CRISPR/Cas9 technology to knock-in GATA2-R398W variant in primary cord blood CD34⁺ cells. Additionally, we introduced specific mutations in SETBP1 and ASXL1 to model distinct premalignant stages of GATA2 deficiency. Through clonal competition and serial transplantation assays, we demonstrated that human CD34<sup>+</sup> cells harboring the GATA2 mutation exhibit significantly reduced fitness in vivo when compete with wild-type cells. Notably, this fitness disadvantage persists even when GATA2 mutations are combined with oncogenic SETBP1 and ASXL1 drivers, underscoring the dominant, deleterious effect of GATA2 deficiency on hematopoietic stem cell function. Functional in vitro analyses revealed that GATA2-R398W mutation impairs cell proliferation, disrupts cell cycle progression, and induces mitotic defects, which may contribute to hematopoietic stem/progenitor cell loss and impaired self-renewal. Transcriptomic profiles of GATA2-mutant cells revealed that these functional defects are associated with reduced HSC self-renewal capacity and upregulation of the pre-aging phenotype. Our work highlights the feasibility of generating a human GATA2 deficiency model suitable for studying the biological consequences of various GATA2 variants and the generation of a platform to test potential phenotype-rescuing therapeutics.
Authors
- Alessandra Giorgetti (ORCID: https://orcid.org/0000-0001-5803-3567)
- Damià Romero–Moya (ORCID: https://orcid.org/0000-0001-9947-3577)
- Arnau Iglesias (ORCID: https://orcid.org/0000-0002-7195-7868)
- Albert Català (ORCID: https://orcid.org/0000-0003-0133-1752)
- Marcin W. Włodarski (ORCID: https://orcid.org/0000-0001-6638-9643)
- Cristina Calvo (ORCID: https://orcid.org/0000-0002-6503-3423)
- Mireya Plass (ORCID: https://orcid.org/0000-0002-9891-2723)
- Julio Castaño (ORCID: https://orcid.org/0000-0002-0712-5856)
- Oskar Marín-Béjar (ORCID: https://orcid.org/0000-0002-6889-6566)
- Marcel Schilling (ORCID: https://orcid.org/0000-0002-3453-7792)
- Anna Bigas (ORCID: https://orcid.org/0000-0003-4801-6899)
- Lorenzo Pasquali (ORCID: https://orcid.org/0000-0003-2423-1826)
- Maximiliano Distefano
- Clara Berenguer-Balaguer
- Francesca De Giorgio (ORCID: https://orcid.org/0000-0002-7108-6978)
- Jéssica González (ORCID: https://orcid.org/0000-0001-7732-1564)
- Joan Pera (ORCID: https://orcid.org/0000-0002-6690-3363)
- Òscar Molina (ORCID: https://orcid.org/0000-0001-7585-4519)
- Eric Torralba-Sales (ORCID: https://orcid.org/0009-0003-7612-2792)
- Maria Magallon-Mosella
Institutions
- Hospital Sant Joan de Déu Barcelona (ES)
- St. Jude Children's Research Hospital (US)
- Universitat Pompeu Fabra (ES)
- Instituto de Salud Carlos III (ES)
- Bellvitge University Hospital (ES)
- Institut d'Investigació Biomédica de Bellvitge (ES)
- Josep Carreras Leukaemia Research Institute (ES)
- Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (ES)
- Ajuntament de L’Hospitalet (ES)
- Centre for Biomedical Network Research on Rare Diseases (ES)
- Department of Physiological Sciences (RU)
- Hospital Del Mar (ES)
- Municipal Institute for Medical Research (ES)
- Institut d'Investigació en Ciències de la Salut Germans Trias i Pujol (ES)
- Hospital del Mar Research Institute (ES)
- Centro de Investigación Biomédica en Red de Cáncer (ES)
- Universitat de Barcelona (ES)
Publication Details
- Journal
- Leukemia
- Published
- 2025-09-15
- DOI
- https://doi.org/10.1038/s41375-025-02771-8
- Citations
- 3
- Primary Topic
- Telomeres, Telomerase, and Senescence
- Type
- article
- Field-Weighted Citation Impact
- 3.49
Funders
- Fundació la Marató de TV3
- Ministerio de Ciencia, Innovación y Universidades
- Centres de Recerca de Catalunya
- European Commission
- Generalitat de Catalunya
- Ministerio de Economía y Competitividad
- Ministerio de Ciencia e Innovación
- José Carreras Leukämie-Stiftung
- Universitat de Barcelona
- Fundación Bancaria Caixa d'Estalvis i Pensions de Barcelona
- Agència de Gestió d'Ajuts Universitaris i de Recerca
- Instituto de Salud Carlos III
- European Regional Development Fund
- Agencia Estatal de Investigación
- European Social Fund