Hematopoietic stem cell gene editing rescues B-cell development in X-linked agammaglobulinemia.
J Allergy Clin Immunol
; 154(1): 195-208.e8, 2024 Jul.
Article
em En
| MEDLINE
| ID: mdl-38479630
ABSTRACT
BACKGROUND:
X-linked agammaglobulinemia (XLA) is an inborn error of immunity that renders boys susceptible to life-threatening infections due to loss of mature B cells and circulating immunoglobulins. It is caused by defects in the gene encoding the Bruton tyrosine kinase (BTK) that mediates the maturation of B cells in the bone marrow and their activation in the periphery. This paper reports on a gene editing protocol to achieve "knock-in" of a therapeutic BTK cassette in hematopoietic stem and progenitor cells (HSPCs) as a treatment for XLA.METHODS:
To rescue BTK expression, this study employed a clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 system that creates a DNA double-strand break in an early exon of the BTK locus and an adeno-associated virus 6 virus that carries the donor template for homology-directed repair. The investigators evaluated the efficacy of the gene editing approach in HSPCs from patients with XLA that were cultured in vitro under B-cell differentiation conditions or that were transplanted in immunodeficient mice to study B-cell output in vivo.RESULTS:
A (feeder-free) B-cell differentiation protocol was successfully applied to blood-mobilized HSPCs to reproduce in vitro the defects in B-cell maturation observed in patients with XLA. Using this system, the investigators could show the rescue of B-cell maturation by gene editing. Transplantation of edited XLA HSPCs into immunodeficient mice led to restoration of the human B-cell lineage compartment in the bone marrow and immunoglobulin production in the periphery.CONCLUSIONS:
Gene editing efficiencies above 30% could be consistently achieved in human HSPCs. Given the potential selective advantage of corrected cells, as suggested by skewed X-linked inactivation in carrier females and by competitive repopulating experiments in mouse models, this work demonstrates the potential of this strategy as a future definitive therapy for XLA.Palavras-chave
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Base de dados:
MEDLINE
Assunto principal:
Células-Tronco Hematopoéticas
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Linfócitos B
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Agamaglobulinemia
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Doenças Genéticas Ligadas ao Cromossomo X
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Edição de Genes
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Tirosina Quinase da Agamaglobulinemia
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article