title: Preclinical Potency and Biodistribution Studies of an AAV 5 Vector Expressing Human Interferon-β (ART-I02) for Local Treatment of Patients with Rheumatoid Arthritis.
abstract: BACKGROUND
  Proof of concept for local gene therapy for the treatment of arthritis with immunomodulatory cytokine interferon beta (IFN-β) has shown promising results in animal models of rheumatoid arthritis (RA). For the treatment of RA patients, we engineered a recombinant adeno-associated serotype 5 vector (rAAV5) encoding human (h)IFN-β under control of a nuclear factor κB promoter (ART-I02).
  
  
  METHODS
  The potency of ART-I02 in vitro as well as biodistribution in vivo in arthritic animals was evaluated to characterize the vector prior to clinical application. ART-I02 expression and bioactivity after transduction was evaluated in fibroblast-like synoviocytes (FLS) from different species. Biodistribution of the vector after local injection was assessed in a rat adjuvant arthritis model through qPCR analysis of vector DNA. In vivo imaging was used to investigate transgene expression and kinetics in a mouse collagen induced arthritis model.
  
  
  RESULTS
  Transduction of RA FLS in vitro with ART-I02 resulted in high expression levels of bioactive hIFN-β. Transduction of FLS from rhesus monkeys, rodents and rabbits with ART-I02 showed high transgene expression, and hIFN-β proved bioactive in FLS from rhesus monkeys. Transgene expression and bioactivity in RA FLS were unaltered in the presence of methotrexate. In vivo, vector biodistribution analysis in rats after intra-articular injection of ART-I02 demonstrated that the majority of vector DNA remained in the joint (>93%). In vivo imaging in mice confirmed local expression of rAAV5 in the knee joint region and demonstrated rapid detectable and sustained expression up until 7 weeks.
  
  
  CONCLUSIONS
  These data show that hIFN-β produced by RA FLS transduced with ART-I02 is bioactive and that intra-articular delivery of rAAV5 drives expression of a therapeutic transgene in the joint, with only limited biodistribution of vector DNA to other tissues, supporting progress towards a phase 1 clinical trial for the local treatment of arthritis in patients with RA.
authors.affiliation: Arthrogen B.V., Amsterdam, the Netherlands; Division of Clinical Immunology and Rheumatology, Academic Medical Center/University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Caroline J
authors.initials: CJ
authors.lastname: Aalbers
authors.affiliation: Arthrogen B.V., Amsterdam, the Netherlands; Division of Clinical Immunology and Rheumatology, Academic Medical Center/University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Lisette
authors.initials: L
authors.lastname: Bevaart
authors.affiliation: Arthrogen B.V., Amsterdam, the Netherlands; Division of Clinical Immunology and Rheumatology, Academic Medical Center/University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Scott
authors.initials: S
authors.lastname: Loiler
authors.affiliation: Arthrogen B.V., Amsterdam, the Netherlands; Division of Clinical Immunology and Rheumatology, Academic Medical Center/University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Karin
authors.initials: K
authors.lastname: de Cortie
authors.affiliation: Center for Cellular and Molecular Therapeutics, The Children's Hospital of Philadelphia, Pennsylvania, United States of America; Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, United States of America.
authors.forename: J Fraser
authors.initials: JF
authors.lastname: Wright
authors.affiliation: University Pierre & Marie Curie, Paris, France; Genethon, Evry, France.
authors.forename: Federico
authors.initials: F
authors.lastname: Mingozzi
authors.affiliation: Arthrogen B.V., Amsterdam, the Netherlands; Division of Clinical Immunology and Rheumatology, Academic Medical Center/University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Paul P
authors.initials: PP
authors.lastname: Tak
authors.affiliation: Arthrogen B.V., Amsterdam, the Netherlands; Division of Clinical Immunology and Rheumatology, Academic Medical Center/University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Margriet J
authors.initials: MJ
authors.lastname: Vervoordeldonk
mesh_terms: D000818:Animals; D001169:Arthritis, Experimental / Q000628:therapy; D001172:Arthritis, Rheumatoid / Q000188:drug therapy / Q000628:therapy*; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D000229:Dependovirus / Q000235:genetics*; D015870:Gene Expression / Q000187:drug effects; D005813:Genes, Synthetic; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics / Q000493:pharmacokinetics / Q000627:therapeutic use*; D006801:Humans; D007270:Injections, Intra-Articular; D016899:Interferon-beta / Q000235:genetics*; D008253:Macaca mulatta; D008297:Male; D008727:Methotrexate / Q000494:pharmacology / Q000627:therapeutic use; D051379:Mice; D008811:Mice, Inbred DBA; D016328:NF-kappa B / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011817:Rabbits; D051381:Rats; D011917:Rats, Inbred Lew; D013047:Specific Pathogen-Free Organisms; D013583:Synovial Membrane / Q000166:cytology / Q000821:virology; D014018:Tissue Distribution; D014161:Transduction, Genetic
chemical_list: D016207:Cytokines; D016328:NF-kappa B; D016899:Interferon-beta; D008727:Methotrexate