Paul D. Robbins

Insulinlike growth factor-I gene therapy applications for cartilage repair

Cartilage function after resurfacing with cell-based transplantation procedures or during the early stages of arthritic disease may be bolstered by the addition of growth factor genes to the transplanted tissue. Insulinlike growth factor-I maintains chondrocyte metabolism in normal cartilage homeostasis and has been shown to improve cartilage healing in vivo. Given the relatively short half-life of insulinlike growth factor-I in biologic systems, however, maintenance of effective concentrations of this peptide has necessitated either very high initial doses or repeated treatment. Delivery of the insulinlike growth factor-I gene, using a deleted adenovirus vector, specifically targeting graftable articular chondrocytes, bone marrow-derived chondroprogenitor cells, or synovial lining cells, may provide more durable insulinlike growth factor-I fluxes to articular tissues. Cultured equine articular chondrocytes, mesenchymal stem cells, synovial explants, and synovial intimal cells were readily transfected with an E1-deleted adenoviral vector containing equine insulinlike growth factor-I coding sequence. Optimal viral concentrations for effective transduction were 100 multiplicities of infection in synoviocytes, 500 multiplicities of infection in chondrocytes, and 1000 multiplicities of infection in mesenchymal stem cells. Production of insulinlike growth factor-I ligand varied from 65 ng/mL to 246 ng/mL in medium from chondrocytes and synovial explants, respectively. For chondrocytes, these concentrations were sufficient to produce significant stimulation of cartilage matrix gene expression and subsequent proteoglycan production. Moreover, cells in infected cultures maintained a chondrocytic phenotype and continued to express elevated insulinlike growth factor-I levels during 28 days of monolayer culture. Minimal synthetic activity, other than insulinlike growth factor-I ligand synthesis, was evident in synovial cultures. These experiments suggest several avenues for insulinlike growth factor-I supplementation of articular cartilage, including preimplantation adenoviral-insulinlike growth factor gene transfer to chondrocytes or chondroprogenitor cells, and direct injection of adenoviral-insulinlike growth factor to transfect the synovial structures in situ.

Gene therapy in inflammatory diseases

List of Contributors. Preface. Christopher H. Evans, Paul D. Robbins: Gene therapy for inflammatory diseases - basic concepts. Sergei S. Makarov: Gene therapy for rheumatoid arthritis pre-clinical studies. Thomas Pap, Ulf Muller-Ladner, Klaus M. Hummel, Renate E. Gay and Steffen Gay: Cartilage erosion in rheumatoid arthritis studies in scid mouse model. Thomas S. Muzzonigro, Richard Kang, J. Reinecke, Peter Wehling, M.C. Wasko and James H. Herndon: Gene therapy for rheumatoid arthritis: clinical studies. Alistair J. Ramsay, Simon P. Hogan, Paul S. Foster and Yelin Xiong: Cytokine gene therapy of allergic airways inflammation. Philip C. Fox and Brian C. OConnell: Gene therapy for inflammatory diseases of the salivary glands. Huang-Ge Zhang, Martin Fleck, Hui-Chen Hsu, Carl K. Edwards III, David T. Curiel, Tong Zhou and John D. Mountz: Gene therapy for management of lupus: correction of Fas and Fas ligand-induced apoptosis in murine disease - therapeutic rational and strategies. Michael K. Shaw, Richard DalCanto and C. Garrison Fathman: Gene therapy for Multiple Sclerosis. Nick Giannoukakis, Massimo Trucco and Paul D. Robbins: Gene therapy for type I Diabetes Mellitus. Jonathan S. Bromberg, Lisa A. DeBruyne, Randall S. Sung and Lihui Qin: Gene transfer to facilitate ransplantation. A. Paul Godillot, Michael Madaio, David B. Weiner and William V. Williams: DNA vaccination as an anti-inflammatory strategy. Masayuki Miyata, Yukio Sato and Reiji Kasukawa: Naked DNA as a gene delivery vehicle in inflammatory diseases. Steven C. Ghivizzani, Eric R. Lechman, Daniel Jaffurs, Zhibao Mi, Richard Kang, Thomas Muzzonigro, Christopher H. Evans and Paul D. Robbins: Animal models of arthritis generated by gene transfer.

Vectors for Gene Transfer to Joints

Gene therapy represents a novel approach for treating joint and bone disorders (Evans et al. 1997; Evans, Ghivizzani, and Robbins 1998; Lattermann et al. 1998). As discussed in other chapters, gene transfer can be used for delivering therapeutic agents to synovium, cartilage, ligaments, tendons, meniscus, intervertebral disc, and bone to block disease progression or to promote repair. In addition, the recent completion of the first gene therapy trial for rheumatoid arthritis (RA) has demonstrated the feasibility of using gene transfer for the treatment of orthopaedic and rheumatologic disorders. The use of gene transfer to deliver a therapeutic agent offers certain advantages over the use of recombinant protein. In particular, the use of genes as therapeutic agents can results in persistent expression locally at the site of disease, bypassing the need for multiple injections and preventing possible side effects associated with systemic administration.