Intraarticular expression of biologically active interleukin 1-receptor-antagonist protein by ex vivo gene transfer.

Abstract

Gene therapy offers a radical different approach to the treatment of arthritis. Here we have demonstrated that two marker genes (lacZ and neo) and cDNA coding for a potentially therapeutic protein (human interleukin 1-receptor-antagonist protein; IRAP or IL-lra) can be delivered, by ex vivo techniques, to the synovial lining of joints; intraarticular expression of IRAP inhibited intraarticular responses to interleukin 1. To achieve this, lapine synoviocytes were first transduced in culture by retroviral infection. The genetically modified synovial cells were then transplanted by intraarticular I jection into the knee joints of rabbits, where they efficiently colonized the synovium. Assay of joint lavages confirmed the in vivo expression of biologically active human IRAP. With allografted cells, IRAP expression was lost by 12 days after transfer. In contrast, autografted synoviocytes continued to express IRAP for -5 weeks. Knee joints expressing human IRAP were protected from the leukocytosis that otherwise follows the intraarticular injection of recombinant human interleukin 1p3. Thus, we report the intraarticular expression and activity of a potentially therapeutic protein by genetransfer technology; these experiments demonstrate the feasibility of treating arthritis and other joint disorders with gene therapy.Arthritis is a chronic, debilitating condition affecting over 30 million Americans (1). Presently incurable, it remains the agent of considerable suffering and economic loss. Therapeutic intervention in arthritis is hindered by a number of factors, including difficulties in targeting drugs to joints. Proteins are particularly vulnerable to this restriction, which is of special concern, as many new agents with considerable antiarthritic potential are proteins. As an alternative to traditional methods of drug delivery, we have suggested the transfer oftherapeutic genes to the synovial lining ofjoints (2,3). Expression of these genes would overcome proteindelivery problems and lead to the intraarticular accumulation of the gene products at the site of disease, with reduced exposure of nontarget organs.Using the rabbit knee joint as a model system, we are therefore developing in vivo and ex vivo methods for delivering genes to joints. This model takes advantage of the similarity in size between the knee joint of the rabbit and the human proximal interphalangeal joint, a common site of rheumatoid arthritis. Moreover, there exists a large body of literature on the biology of the rabbit's knee, including well-established methods for synovial cell culture (e.g., refs. 4-6). Here we report the transfer to synovium of two marker genes and one potentially therapeutic gene by an ex vivo approach. Intraarticular expression of the interleukin 1-re-The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. (rhIL-1,8). These results demonstrate the feasibility of ...

Intraarticular induction of interleukin‐1β expression in the adult mouse, with resultant temporomandibular joint pathologic changes, dysfunction, and pain

“…The lack of inflammatory cells or inflamed pannus has also been previously described in the rat model of iodoacetate-induced OA, where an inflammatory process that is not associated with cell infiltration may underlie the development of OA, and there is pain behavior related to the joints (29). Conversely, previous studies using IL-1␤ in rodent models of arthritis have shown RA-like pathologic development in the joint for up to 1 month following intraarticular cytokine injection of methylated bovine serum albumin (10) or ex vivo administration of IL-1-transduced synovial cells (30,31), which resolved shortly thereafter. In contrast to these short-term models, the Col1-IL-1␤ XAT -transgenic mouse may serve as a model for long-term studies of arthritis.…”

Intra-articular delivery of a herpes simplex virus IL-1Ra gene vector reduces inflammation in a rabbit model of arthritis

“…28 Plasmid P41ICPO-IL-1Ra: This plasmid was constructed in the same manner as p41ICP0-TNFSRexcept that the IL1Ra cDNA fused to the SV40 polyadenylation sequence was inserted into p41ICP0lacZ in place of the TNFSR cDNA. 3 Vectors S/0-TNFSR and T/0-sTNF␣R: The TNFSR recombination plasmid described above was introduced into the genome of SOZ.1 using the previously described PacI recombination strategy. 26 Briefly, PacI endonuclease digested SOZ.1 genomic DNA was co-transfected along with the recombination plasmid p41ICP0-TNFSRinto 7b cells using standard calcium phosphate methods.…”

“…For this procedure, cells were surgically harvested from the synovial lining of the joint, cultured and transduced by a retroviral vector in vitro, and the genetically modified cells injected into the joint space to recolonize the synovium. 3,4 Ex vivo delivery of the interleukin-1 receptor antagonist cDNA to the joints of rabbits with experimental arthritis was found to have significant therapeutic benefit. 5 More recently, a phase I clinical trial was implemented to evaluate the safety and feasibility of using ex vivo gene delivery in the treatment of RA.…”

Induction of apoptosis in the rheumatoid synovium by Fas ligand gene transfer

“…Several recent studies have been successful in introducing genes into the synovium of the knee joints of rabbits by both ex vivo 30,31 and in vitro 32,33 methods. One potentially therapeutic gene encoding the human interleukin-1 (IL-1) receptor antagonist has been tested in some detail.…”