Disease- and cell-type-specific transcriptional targeting of vectors for osteoarthritis gene therapy: further development of a clinical canine model

• Published in: Rheumatology (Oxford, England) Vol. 44; no. 6; pp. 735 - 743
• Main Authors: Campbell, S. E., Bennett, D., Nasir, L., Gault, E. A., Argyle, D. J.
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01.06.2005; Oxford Publishing Limited (England)
• Subjects: Animals; Base Sequence; Biological and medical sciences; Canine model; Cell Line, Tumor; Collagen Type XI - genetics; Disease Models, Animal; Diseases of the osteoarticular system; Dogs; Gene Deletion; Gene therapy; Genetic Therapy - methods; Genetic Vectors - genetics; High Mobility Group Proteins - genetics; Interleukin-1 - genetics; Introns - genetics; Matrix Metalloproteinase 9 - genetics; Medical sciences; Mice; Miscellaneous. Osteoarticular involvement in other diseases; Mutation - genetics; NF-kappa B - genetics; Osteoarthritis; Osteoarthritis - genetics; Osteoarthritis - therapy; Promoter Regions, Genetic - genetics; SOX9 Transcription Factor; Transcription; Transcription Factors - genetics; Transcription, Genetic - genetics; Tumor Necrosis Factor-alpha - genetics; Fissipedia; Carnivora; Targeting; Transcription; Diseases of the osteoarticular system; Cuspid; Vertebrata; Canine model; Mammalia; Treatment; Animal; Arthropathy; Degenerative disease; Models; Gene therapy; Osteoarthritis; Dog; Vector
• ISSN: 1462-0324; 1462-0332
• DOI: 10.1093/rheumatology/keh590

Objectives. The potential for undesirable systemic effects related to constitutive expression of certain therapeutic transgenes may be limited through the development of transcriptionally targeted disease- and cell-type-specific vectors. The objective of this study was to analyse the canine matrix metalloproteinase-9 (MMP-9) promoter and deletion constructs for its ability to drive expression in response to pro-inflammatory cytokines (interleukin-1β and tumour necrosis factor-α). Methods. Initial analysis of MMP-9 deletion constructs was made using a luciferase reporter system. The promoter was subsequently engineered to incorporate multiple NF-κB sites. In parallel experiments we used the mouse collagen type XI promoter to study cell-type-specific promoter activity in chondrocyte-specific cells (SW1353) and undifferentiated chondroprogenitor cells (ATDC5). Results. Incorporation of multiple NF-κB sites into the MMP-9 promoter enhanced activity while maintaining disease specificity. Further, manipulation of the mouse collagen type XI (mColXI) promoter by the incorporation of SOX9 enhancer sites downstream of a reporter gene, increased gene activity while maintaining cell type specificity. Conclusions. Manipulation of promoter and enhancer regions can improve transcriptionally targeted genes. A combination of these systems, in the context of the canine model, has the potential to improve the safety of osteoarthritis gene therapy vectors.