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Ribozymes as tools for therapeutic target validation in arthritis.
- Source :
-
Journal of immunology (Baltimore, Md. : 1950) [J Immunol] 2000 Jul 01; Vol. 165 (1), pp. 493-8. - Publication Year :
- 2000
-
Abstract
- In this paper we describe a method for validating therapeutic gene targets in arthritic disease. Ribozymes are catalytic oligonucleotides capable of highly sequence-specific cleavage of RNA. We designed ribozymes that cleave the mRNA encoding stromelysin, a matrix metalloproteinase implicated in cartilage catabolism. Ribozymes were initially screened in cultured fibroblasts to identify sites in the mRNA that were accessible for binding and cleavage. Accessible sites for ribozyme binding were found in various regions of the mRNA, including the 5' untranslated region, the coding region, and the 3' untranslated region. Several ribozymes that mediated sequence-specific and dose-dependent inhibition of stromelysin expression were characterized. Site selection in cell culture was predictive of in vivo bioactivity. An assay for measuring cartilage catabolism in rabbit articular cartilage explants was developed. Ribozymes inhibited IL-1-stimulated stromelysin mRNA expression in articular cartilage explants, yet failed to inhibit proteoglycan degradation. This indicated that up-regulation of stromelysin was not essential for IL-1-induced cartilage catabolism. Broad applications of this approach in therapeutic target validation are discussed.
- Subjects :
- Animals
Arthritis genetics
Arthritis metabolism
Cartilage, Articular enzymology
Cartilage, Articular metabolism
Cells, Cultured
Dose-Response Relationship, Drug
Female
Fibroblasts enzymology
Humans
Hydrolysis
Injections, Intra-Articular
Male
Matrix Metalloproteinase 3 genetics
Matrix Metalloproteinase 3 metabolism
Matrix Metalloproteinase 3 physiology
Matrix Metalloproteinase Inhibitors
Organ Culture Techniques
RNA, Catalytic administration & dosage
RNA, Catalytic metabolism
Rabbits
Reproducibility of Results
Substrate Specificity
Synovial Membrane enzymology
Synovial Membrane metabolism
Arthritis enzymology
Arthritis therapy
Gene Targeting methods
RNA, Catalytic therapeutic use
Subjects
Details
- Language :
- English
- ISSN :
- 0022-1767
- Volume :
- 165
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Journal of immunology (Baltimore, Md. : 1950)
- Publication Type :
- Academic Journal
- Accession number :
- 10861088
- Full Text :
- https://doi.org/10.4049/jimmunol.165.1.493