A long-term study of AAV gene therapy in dogs with hemophilia A identifies clonal expansions of transduced liver cells

Giang N. Nguyen; John K. Everett; Samita Kafle; Aoife M. Roche; Hayley E. Raymond; Jacob Leiby; Christian Wood; Charles Antoine Assenmacher; Elizabeth P. Merricks; C. Tyler Long; Haig H. Kazazian; Timothy C. Nichols; Frederic D. Bushman; Denise E. Sabatino

doi:10.1038/s41587-020-0741-7

A long-term study of AAV gene therapy in dogs with hemophilia A identifies clonal expansions of transduced liver cells

Giang N. Nguyen, John K. Everett, Samita Kafle, Aoife M. Roche, Hayley E. Raymond, Jacob Leiby, Christian Wood, Charles Antoine Assenmacher, Elizabeth P. Merricks, C. Tyler Long, Haig H. Kazazian, Timothy C. Nichols, Frederic D. Bushman, Denise E. Sabatino

School of Medicine

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

Nine dogs with hemophilia A were treated with adeno-associated viral (AAV) gene therapy and followed for up to 10 years. Administration of AAV8 or AAV9 vectors expressing canine factor VIII (AAV-cFVIII) corrected the FVIII deficiency to 1.9–11.3% of normal FVIII levels. In two of nine dogs, levels of FVIII activity increased gradually starting about 4 years after treatment. None of the dogs showed evidence of tumors or altered liver function. Analysis of integration sites in liver samples from six treated dogs identified 1,741 unique AAV integration events in genomic DNA and expanded cell clones in five dogs, with 44% of the integrations near genes involved in cell growth. All recovered integrated vectors were partially deleted and/or rearranged. Our data suggest that the increase in FVIII protein expression in two dogs may have been due to clonal expansion of cells harboring integrated vectors. These results support the clinical development of liver-directed AAV gene therapy for hemophilia A, while emphasizing the importance of long-term monitoring for potential genotoxicity.

Original language	English (US)
Pages (from-to)	47-55
Number of pages	9
Journal	Nature biotechnology
Volume	39
Issue number	1
DOIs	https://doi.org/10.1038/s41587-020-0741-7
State	Published - Jan 2021

ASJC Scopus subject areas

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology
Molecular Medicine
Biomedical Engineering

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Nguyen, G. N., Everett, J. K., Kafle, S., Roche, A. M., Raymond, H. E., Leiby, J., Wood, C., Assenmacher, C. A., Merricks, E. P., Long, C. T., Kazazian, H. H., Nichols, T. C., Bushman, F. D., & Sabatino, D. E. (2021). A long-term study of AAV gene therapy in dogs with hemophilia A identifies clonal expansions of transduced liver cells. Nature biotechnology, 39(1), 47-55. https://doi.org/10.1038/s41587-020-0741-7

Nguyen, GN, Everett, JK, Kafle, S, Roche, AM, Raymond, HE, Leiby, J, Wood, C, Assenmacher, CA, Merricks, EP, Long, CT, Kazazian, HH, Nichols, TC, Bushman, FD & Sabatino, DE 2021, 'A long-term study of AAV gene therapy in dogs with hemophilia A identifies clonal expansions of transduced liver cells', Nature biotechnology, vol. 39, no. 1, pp. 47-55. https://doi.org/10.1038/s41587-020-0741-7

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title = "A long-term study of AAV gene therapy in dogs with hemophilia A identifies clonal expansions of transduced liver cells",

abstract = "Nine dogs with hemophilia A were treated with adeno-associated viral (AAV) gene therapy and followed for up to 10 years. Administration of AAV8 or AAV9 vectors expressing canine factor VIII (AAV-cFVIII) corrected the FVIII deficiency to 1.9–11.3% of normal FVIII levels. In two of nine dogs, levels of FVIII activity increased gradually starting about 4 years after treatment. None of the dogs showed evidence of tumors or altered liver function. Analysis of integration sites in liver samples from six treated dogs identified 1,741 unique AAV integration events in genomic DNA and expanded cell clones in five dogs, with 44% of the integrations near genes involved in cell growth. All recovered integrated vectors were partially deleted and/or rearranged. Our data suggest that the increase in FVIII protein expression in two dogs may have been due to clonal expansion of cells harboring integrated vectors. These results support the clinical development of liver-directed AAV gene therapy for hemophilia A, while emphasizing the importance of long-term monitoring for potential genotoxicity.",

author = "Nguyen, {Giang N.} and Everett, {John K.} and Samita Kafle and Roche, {Aoife M.} and Raymond, {Hayley E.} and Jacob Leiby and Christian Wood and Assenmacher, {Charles Antoine} and Merricks, {Elizabeth P.} and Long, {C. Tyler} and Kazazian, {Haig H.} and Nichols, {Timothy C.} and Bushman, {Frederic D.} and Sabatino, {Denise E.}",

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A long-term study of AAV gene therapy in dogs with hemophilia A identifies clonal expansions of transduced liver cells

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