title: Safety of AAV factor IX peripheral transvenular gene delivery to muscle in hemophilia B dogs.
abstract: Muscle represents an attractive target tissue for adeno-associated virus (AAV) vector-mediated gene transfer for hemophilia B (HB). Experience with direct intramuscular (i.m.) administration of AAV vectors in humans showed that the approach is safe but fails to achieve therapeutic efficacy. Here, we present a careful evaluation of the safety profile (vector, transgene, and administration procedure) of peripheral transvenular administration of AAV-canine factor IX (cFIX) vectors to the muscle of HB dogs. Vector administration resulted in sustained therapeutic levels of cFIX expression. Although all animals developed a robust antibody response to the AAV capsid, no T-cell responses to the capsid antigen were detected by interferon (IFN)-gamma enzyme-linked immunosorbent spot (ELISpot). Interleukin (IL)-10 ELISpot screening of lymphocytes showed reactivity to cFIX-derived peptides, and restimulation of T cells in vitro in the presence of the identified cFIX epitopes resulted in the expansion of CD4(+)FoxP3(+)IL-10(+) T-cells. Vector administration was not associated with systemic inflammation, and vector spread to nontarget tissues was minimal. At the local level, limited levels of cell infiltrates were detected when the vector was administered intravascularly. In summary, this study in a large animal model of HB demonstrates that therapeutic levels of gene transfer can be safely achieved using a novel route of intravascular gene transfer to muscle.
authors.affiliation: Division of Hematology and Center for Cellular and Molecular Therapeutics, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Virginia
authors.initials: V
authors.lastname: Haurigot
authors.forename: Federico
authors.initials: F
authors.lastname: Mingozzi
authors.forename: George
authors.initials: G
authors.lastname: Buchlis
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Hui
authors.forename: Yifeng
authors.initials: Y
authors.lastname: Chen
authors.forename: Etiena
authors.initials: E
authors.lastname: Basner-Tschakarjan
authors.forename: Valder R
authors.initials: VR
authors.lastname: Arruda
authors.forename: Antoneta
authors.initials: A
authors.lastname: Radu
authors.forename: Helen G
authors.initials: HG
authors.lastname: Franck
authors.forename: J Fraser
authors.initials: JF
authors.lastname: Wright
authors.forename: Shangzhen
authors.initials: S
authors.lastname: Zhou
authors.forename: Hansell H
authors.initials: HH
authors.lastname: Stedman
authors.forename: Dwight A
authors.initials: DA
authors.lastname: Bellinger
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Nichols
authors.forename: Katherine A
authors.initials: KA
authors.lastname: High
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D004285:Dogs; D005164:Factor IX / Q000235:genetics* / Q000378:metabolism; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000009:adverse effects* / Q000235:genetics*; D002836:Hemophilia B / Q000378:metabolism / Q000628:therapy*; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007371:Interferon-gamma / Q000378:metabolism; D016753:Interleukin-10 / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000473:pathology
chemical_list: D007074:Immunoglobulin G; D016753:Interleukin-10; D007371:Interferon-gamma; D005164:Factor IX