Preclinical Potency and Biodistribution Studies of an AAV 5 Vector Expressing Human Interferon-β (ART-I02) for Local Treatment of Patients with Rheumatoid Arthritis

PLoS One. 2015 Jun 24;10(6):e0130612. doi: 10.1371/journal.pone.0130612. eCollection 2015.


Introduction: 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.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Arthritis, Experimental / therapy
  • Arthritis, Rheumatoid / drug therapy
  • Arthritis, Rheumatoid / therapy*
  • Cells, Cultured
  • Cytokines / biosynthesis
  • Dependovirus / genetics*
  • Gene Expression / drug effects
  • Genes, Synthetic
  • Genetic Therapy*
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics
  • Genetic Vectors / pharmacokinetics
  • Genetic Vectors / therapeutic use*
  • Humans
  • Injections, Intra-Articular
  • Interferon-beta / genetics*
  • Macaca mulatta
  • Male
  • Methotrexate / pharmacology
  • Methotrexate / therapeutic use
  • Mice
  • Mice, Inbred DBA
  • NF-kappa B / genetics*
  • Promoter Regions, Genetic / genetics*
  • Rabbits
  • Rats
  • Rats, Inbred Lew
  • Specific Pathogen-Free Organisms
  • Synovial Membrane / cytology
  • Synovial Membrane / virology
  • Tissue Distribution
  • Transduction, Genetic


  • Cytokines
  • NF-kappa B
  • Interferon-beta
  • Methotrexate

Grant support

This research was supported by Arthrogen. Co-authors Lisette Bevaart and Margriet J. Vervoordeldonk are currently employed by Arthrogen BV and receive a salary. PP Tak is inventor on the patent “AAV vectors for in vivo gene therapy of rheumatoid arthritis” (EP 2 322 638 A1) but does not receive any financial compensation for this patent from Arthrogen or any other institution. The specific roles of the authors are articulated in the ‘author contributions’ section.