Optimization of scAAVIL-1ra in vitro and in vivo to deliver high levels of therapeutic protein for treatment of osteoarthritis

Laurie R. Goodrich, Jennifer N. Phillips, C. Wayne McIlwraith, Stacey B. Foti, Joshua C. Grieger, Steven J. Gray, R. Jude Samulski

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Osteoarthritis (OA) affects over 40 million people annually. We evaluated interleukin-1 receptor antagonist (IL-1ra) gene transfer in an equine model based on IL-1ra protein therapy which inhibits inflammation through blocking IL-1. Using the self-complementary adeno-associated virus (scAAV)IL-1ra equine gene as a starting construct, we optimized the transgene cassette by analyzing promoters (cytomegalovirus (CMV) versus chicken β-actin hybrid (CBh)), coding sequences (optimized versus unoptimized), vector capsid (serotype 2 versus chimeric capsid), and biological activity in vitro. AAV serotypes 2 and 2.5 CMV scAAVoptIL-1ra were tested in equine joints. We evaluated two doses of scAAVIL-1ra, scAAVGFP, and saline. We developed a novel endoscopy procedure and confirmed vector-derived transgene expression (GFP) in chondrocytes 6 months post-injection. AAVIL-1ra therapeutic protein levels were 200-800 ng/ml of synovial fluid over 23 and 186 days, respectively. No evidence of intra-articular toxicity was detected and no vector genomes were found in contralateral joints based on GFP fluorescence microscopy and quantitative PCR. Finally, we assayed vector-derived IL-1ra activity based on functional assays which supported anti-inflammatory activity of our protein. These studies represent the first large animal intra-articular gene transfer approach with a therapeutic gene using scAAV and demonstrate high levels of protein production over extended time supporting further clinical investigation using scAAV gene therapy for OA.

Original languageEnglish (US)
Article numbere70
JournalMolecular Therapy - Nucleic Acids
Volume2
DOIs
StatePublished - Jun 17 2013

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Osteoarthritis
Dependovirus
Interleukin-1 Receptors
Joints
Horses
Capsid
Cytomegalovirus
Transgenes
Genes
Proteins
Interleukin 1 Receptor Antagonist Protein
Synovial Fluid
Therapeutics
Chondrocytes
Interleukin-1
Fluorescence Microscopy
Genetic Therapy
Endoscopy
Actins
Chickens

Keywords

  • IL-1ra
  • Intraarticular
  • Joints and arthroscopy
  • ScAAV

ASJC Scopus subject areas

  • Molecular Medicine
  • Drug Discovery

Cite this

Optimization of scAAVIL-1ra in vitro and in vivo to deliver high levels of therapeutic protein for treatment of osteoarthritis. / Goodrich, Laurie R.; Phillips, Jennifer N.; McIlwraith, C. Wayne; Foti, Stacey B.; Grieger, Joshua C.; Gray, Steven J.; Samulski, R. Jude.

In: Molecular Therapy - Nucleic Acids, Vol. 2, e70, 17.06.2013.

Research output: Contribution to journalArticle

Goodrich, Laurie R. ; Phillips, Jennifer N. ; McIlwraith, C. Wayne ; Foti, Stacey B. ; Grieger, Joshua C. ; Gray, Steven J. ; Samulski, R. Jude. / Optimization of scAAVIL-1ra in vitro and in vivo to deliver high levels of therapeutic protein for treatment of osteoarthritis. In: Molecular Therapy - Nucleic Acids. 2013 ; Vol. 2.
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