Factor VIII delivered by haematopoietic stem cell-derived B cells corrects the phenotype of haemophilia A mice

Ali Ramezani; Lynnsey A. Zweier-Renn; Robert G. Hawley

doi:10.1160/TH10-11-0725

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2011; 105(04): 676-687
DOI: 10.1160/TH10-11-0725

Animal Models

Schattauer GmbH

Factor VIII delivered by haematopoietic stem cell-derived B cells corrects the phenotype of haemophilia A mice

Ali Ramezani

¹Department of Anatomy and Regenerative Biology, The George Washington University, Washington, District of Columbia, USA

,

Lynnsey A. Zweier-Renn

¹Department of Anatomy and Regenerative Biology, The George Washington University, Washington, District of Columbia, USA

²Graduate Program in Biochemistry and Molecular Genetics, The George Washington University, Washington, District of Columbia, USA

,

Robert G. Hawley

¹Department of Anatomy and Regenerative Biology, The George Washington University, Washington, District of Columbia, USA

²Graduate Program in Biochemistry and Molecular Genetics, The George Washington University, Washington, District of Columbia, USA

› Author Affiliations

Abstract

Summary

The main impediments to clinical application of haematopoietic stem cell (HSC) gene therapy for treatment of haemophilia A are the bone marrow transplant-related risks and the potential for insertional muta-genesis caused by retroviral vectors. To circumvent these limitations, we have adapted a non-myeloablative conditioning regimen and directed factor VIII (FVIII) protein synthesis to B lineage cells using an insulated lentiviral vector containing an immunoglobulin heavy chain enhancer-promoter. Transplantation of lentiviral vector-modified HSCs resulted in therapeutic levels of FVIII in the circulation of all transplanted mice for the duration of the study (six months). Immunostaining of spleen cells showed that the majority of FVIII was synthesised by B220⁺ B cells and CD138⁺ plasma cells. Subsequent challenge with recombinant FVIII elicited at most a minor anti-FVIII antibody response, demonstrating in-duction of immune hyporesponsiveness. All transplant recipients exhibited clot formation and survived tail clipping, indicating correction of their haemophilic phenotype. Therapeutic levels of FVIII could be transferred to secondary recipients by bone marrow transplantation, confirming gene transfer into long-term repopulating HSCs. Moreover, short-term therapeutic FVIII levels could also be achieved in secondary recipients by adoptive transfer of HSC-derived splenic B cells. Our findings support pursuit of B cell-directed protein delivery as a potential clinical approach to treat haemophilia A and other disorders correctable by systemically distributed proteins.

Keywords

Haemophilia A - animal models - haematopoietic stem cell gene therapy - B cell-targeted transgene expression - bioengineered factor VIII

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References

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