Abstract
Pre-clinical studies in mice and haemophilic dogs have shown that introduction of an adeno-associated viral (AAV) vector encoding blood coagulation factor IX (F.IX) into skeletal muscle results in sustained expression of F.IX at levels sufficient to correct the haemophilic phenotype1,2. On the basis of these data and additional pre-clinical studies demonstrating an absence of vector-related toxicity, we initiated a clinical study of intramuscular injection of an AAV vector expressing human F.IX in adults with severe haemophilia B. The study has a dose-escalation design, and all patients have now been enrolled in the initial dose cohort (2×1011 vg/kg). Assessment in the first three patients of safety and gene transfer and expression show no evidence of germline transmission of vector sequences or formation of inhibitory antibodies against F.IX. We found that the vector sequences are present in muscle by PCR and Southern-blot analyses of muscle biopsies and we demonstrated expression of F.IX by immunohistochemistry. We observed modest changes in clinical endpoints including circulating levels of F.IX and frequency of F.IX protein infusion. The evidence of gene expression at low doses of vector suggests that dose calculations based on animal data may have overestimated the amount of vector required to achieve therapeutic levels in humans, and that the approach offers the possibility of converting severe haemophilia B to a milder form of the disease.
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Acknowledgements
We thank A. Radu for assistance with immunohistochemical staining of muscle; M. Tanzer for assistance with coagulation assays; D. Leonard for assistance with PCR assays on human samples; the Nucleic Acid/Protein Research Core Facility at CHOP; R. Barth and M. King for assistance with ultrasound; and S.G. Madison for clinical support. This work was supported by National Institutes of Health Grants R01 HL53682 to M.A.K. and R01 HL53668, R01 HL61921, P50 HL54500 and a grant from the Hoxie Harrison-Smith Foundation to K.A.H. The work was also supported in part by NIH grant M01 RR00070 to the GCRC at Stanford University, by NIH grant M01 RR00240 to the GCRC at The Children's Hospital of Philadelphia and by Avigen.
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Kay, M., Manno, C., Ragni, M. et al. Evidence for gene transfer and expression of factor IX in haemophilia B patients treated with an AAV vector. Nat Genet 24, 257–261 (2000). https://doi.org/10.1038/73464
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DOI: https://doi.org/10.1038/73464
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