Jean-Noel Gouze

Exogenous glucosamine globally protects chondrocytes from the arthritogenic effects of IL-1β

The effects of exogenous glucosamine on the biology of articular chondrocytes were determined by examining global transcription patterns under normal culture conditions and following challenge with IL-1β. Chondrocytes isolated from the cartilage of rats were cultured in several flasks either alone or in the presence of 20 mM glucosamine. Six hours later, one-half of the cultures of each group were challenged with 10 ng/ml IL-1β. Fourteen hours after this challenge, RNA was extracted from each culture individually and used to probe microarray chips corresponding to the entire rat genome. Glucosamine alone had no observable stimulatory effect on the transcription of primary cartilage matrix genes, such as aggrecan, collagen type II, or genes involved in glycosaminoglycan synthesis; however, glucosamine proved to be a potent, broad-spectrum inhibitor of IL-1β. Of the 2,813 genes whose transcription was altered by IL-1β stimulation (P < 0.0001), glucosamine significantly blocked the response in 2,055 (~73%). Glucosamine fully protected the chondrocytes from IL-1-induced expression of inflammatory cytokines, chemokines, and growth factors as well as proteins involved in prostaglandin E2 and nitric oxide synthesis. It also blocked the IL-1-induced expression of matrix-specific proteases such as MMP-3, MMP-9, MMP-10, MMP-12, and ADAMTS-1. The concentrations of IL-1 and glucosamine used in these assays were supraphysiological and were not representative of the arthritic joint following oral consumption of glucosamine. They suggest, however, that the potential benefit of glucosamine in osteoarthritis is not related to cartilage matrix biosynthesis, but is more probably related to its ability to globally inhibit the deleterious effects of IL-1β signaling. These results suggest that glucosamine, if administered effectively, may indeed have anti-arthritic properties, but primarily as an anti-inflammatory agent.

Intra‐articular gene delivery and expression of interleukin‐1Ra mediated by self‐complementary adeno‐associated virus

The adeno‐associated virus (AAV) has many safety features that favor its use in the treatment of arthritic conditions; however, the conventional, single‐stranded vector is inefficient for gene delivery to fibroblastic cells that primarily populate articular tissues. This has been attributed to the inability of these cells to convert the vector to a double‐stranded form. To overcome this, we evaluated double‐stranded self‐complementary (sc) AAV as a vehicle for intra‐articular gene delivery.

scAAV-Mediated Gene Transfer of Interleukin 1-Receptor Antagonist to Synovium and Articular Cartilage in Large Mammalian Joints

With the long-term goal of developing a gene-based treatment for osteoarthritis (OA), we performed studies to evaluate the equine joint as a model for adeno-associated virus (AAV)-mediated gene transfer to large, weight-bearing human joints. A self-complementary AAV2 vector containing the coding regions for human interleukin-1-receptor antagonist (hIL-1Ra) or green fluorescent protein was packaged in AAV capsid serotypes 1, 2, 5, 8 and 9. Following infection of human and equine synovial fibroblasts in culture, we found that both were only receptive to transduction with AAV1, 2 and 5. For these serotypes, however, transgene expression from the equine cells was consistently at least 10-fold higher. Analyses of AAV surface receptor molecules and intracellular trafficking of vector genomes implicate enhanced viral uptake by the equine cells. Following delivery of 1 × 1011 vector genomes of serotypes 2, 5 and 8 into the forelimb joints of the horse, all three enabled hIL-1Ra expression at biologically relevant levels and effectively transduced the same cell types, primarily synovial fibroblasts and, to a lesser degree, chondrocytes in articular cartilage. These results provide optimism that AAV vectors can be effectively adapted for gene delivery to large human joints affected by OA.

Perspectives on the use of gene therapy for chronic joint diseases.

Advances in molecular and cellular biology have identified a wide variety of proteins including targeted cytokine inhibitors, immunomodulatory proteins, cytotoxic mediators, angiogenesis inhibitors, and intracellular signalling molecules that could be of great benefit in the treatment of chronic joint diseases, such as osteo- and rheumatoid arthritis. Unfortunately, protein-based drugs are difficult to administer effectively. They have a high rate of turnover, requiring frequent readministration, and exposure in non-diseased tissue can lead to serious side effects. Gene transfer technologies offer methods to enhance the efficacy of protein-based therapies, enabling the body to produce these molecules locally at elevated levels for extended periods. The proof of concept of gene therapies for arthritis has been exhaustively demonstrated in multiple laboratories and in numerous animal models. This review attempts to condense these studies and to discuss the relative benefits and limitations of the methods proposed and to discuss the challenges toward translating these technologies into clinical realities.

Interleukin-1 in rheumatoid arthritis : Its inhibition by IL-1Ra and Anakinra

Objective: To review the biology of interleukin-1 (IL-1) in the pathogenesis of rheumatoid arthritis (RA), as well as the biology of its natural inhibitor, IL receptor antagonist (IL-1Ra), and the clinical efficacy and safety of the recombinant form, anakinra. Data Sources: A MEDLINE search (1966–January 2007) of English-language articles was conducted using the key words anakinra, arthritis, clinical trial, interleukin-1 receptor antagonist, and Kineret. Study Selection and Data Extraction: Over 79 research articles and literature reviews were used to compile a discussion of the biology of IL-1 and IL-1Ra. Ten of these articles were selected to discuss the clinical safety and efficacy of anakinra. Data Synthesis: In RA, IL-1 primarily acts locally to mediate erosion of cartilage and bone. IL-1Ra serves to modulate its activity through competitive inhibition of cellular receptors. Administration of anakinra to animals with experimental arthritis reduced inflammation and joint damage. In clinical trials, anakinra was reasonably well tolerated; however, injection site reactions were frequent and there was a slight increased risk of serious infection. Alone or in combination with methotrexate, anakinra significantly reduced the symptoms and clinical signs of RA at the American College of Rheumatology 20% response level. However, no additive benefit was observed following coadministration with etanercept, a soluble tumor necrosis factor antagonist, and anakinra had no beneficial effect in patients that failed treatment with etanercept. Conclusions: Laboratory studies have indicated that IL-1 is primarily responsible for cartilage destruction and bone erosion in RA. The selective inhibition of IL-1 through administration of anakinra may offer symptomatic relief of RA in some patients.