title: Persistent expression of canine factor IX in hemophilia B canines.
abstract: We previously demonstrated that direct intramuscular injection of recombinant adeno-associated virus (rAAV) carrying the human FIX (hFIX) cDNA can safely be administered to hemophilic B canines and express human factor IX protein; however, the functional activity of the hFIX protein could not be assessed due to anti-human FIX antibody (inhibitor) formation. To test the therapeutic efficacy of rAAV in hemophilic dogs, rAAV type 2 (rAAV2) carrying canine FIX (cFIX) cDNA was injected into the skeletal muscle of two dogs at doses of 1012-13particles. Circulating cFIX protein levels were maintained for 1 year at levels of 1-2% of normal. Hemostatic correction (WBCT and APTT) paralleled plasma FIX antigen levels. Both dogs still required plasma infusion for spontaneous and traumatic bleeding events. Inhibitors to cFIX protein were not detected in either animal by Bethesda assay. Neutralizing antibodies directed against AAV-2 capsid were pronounced and persistent. Vector DNA and mRNA transcripts were detected only at the injected skeletal muscle tissue. Analysis of both high and low molecular weight DNA identified both replicative episomal and integrated AAV species. These results demonstrate that persistent secretion of the FIX transgene protein, necessary for successful gene therapy of hemophilia B, can be achieved using the parvovirus-based rAAV vector
authors.affiliation: UNC Gene Therapy Center, University of North Carolina, Chapel Hill, NC 27599, USA.
authors.forename: H
authors.initials: H
authors.lastname: Chao
authors.forename: R
authors.initials: R
authors.lastname: Samulski
authors.forename: D
authors.initials: D
authors.lastname: Bellinger
authors.forename: P
authors.initials: P
authors.lastname: Monahan
authors.forename: T
authors.initials: T
authors.lastname: Nichols
authors.forename: C
authors.initials: C
authors.lastname: Walsh
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D001777:Blood Coagulation; D015139:Blotting, Southern; D004247:DNA / Q000032:analysis; D000229:Dependovirus / Q000235:genetics / Q000276:immunology; D004283:Dog Diseases / Q000628:therapy*; D004285:Dogs; D005164:Factor IX / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D002836:Hemophilia B / Q000628:therapy* / Q000662:veterinary*; D006801:Humans; D007273:Injections, Intramuscular; D018482:Muscle, Skeletal / Q000821:virology; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
chemical_list: D000914:Antibodies, Viral; D012333:RNA, Messenger; D005164:Factor IX; D004247:DNA