Hematopoietic stem cell gene therapy for the treatment of X-linked agammaglobulinemia

Christopher R Luthers; Annika Mittelhauser; Aurelien Colamartino; Xiaomeng Wu; Samuel Cirigliano; Joseph D Long; Julie M Sanchez; Zulema Romero; Donald B Kohn

doi:10.1016/j.omtm.2025.101555

Hematopoietic stem cell gene therapy for the treatment of X-linked agammaglobulinemia

Mol Ther Methods Clin Dev. 2025 Aug 12;33(3):101555. doi: 10.1016/j.omtm.2025.101555. eCollection 2025 Sep 11.

Authors

Affiliations

¹ Molecular Biology Interdepartmental Program, University of California, Los Angeles, Los Angeles, CA 90095, USA.
² Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ Department of Pediatrics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.

Abstract

X-linked agammaglobulinemia (XLA) is a rare inborn error of immunity caused by loss-of-function mutations in the gene encoding Bruton's tyrosine kinase (BTK). XLA patients lack mature B cells and have negligible antibody levels, leaving them susceptible to recurrent bacterial and chronic viral infections. Autologous hematopoietic stem cell gene therapy with gene-corrected HSC may serve as a promising treatment of XLA; this therapy would provide a one-time cure and would replace lifelong immunoglobulin replacement therapy. Due to the requirement of strict physiological regulation of BTK gene expression, a site-specific editing strategy was designed to insert a BTK cDNA transgene directly into its endogenous locus. To study the effectiveness of this therapy, murine lineage-negative hematopoietic cells from a murine model of XLA were edited using CRISPR-Cas9/rAAV6 then transplanted into recipient XLA mice. Myeloablated XLA mice that received transplantation of Btk-corrected Lin- cells displayed high levels of engraftment, significant increases in their B cell levels, increased production of various immunoglobulins, improved B cell development in the bone marrow, increased B cell receptor diversity, and the ability to produce antigen-specific antibodies following immunization. Collectively, we have modeled a gene therapy strategy in a disease model of XLA and extensively validated the site-specific genome editing approach.

Keywords: B cell development; Bruton’s tyrosine kinase; CRISPR-Cas9; X-linked agammaglobulinemia; adeno-associated virus; gene therapy; hematopoietic stem cells; immunodeficiency; immunoglobulins; site-specific genome editing.