RNAi reduces expression and intracellular retention of mutant cartilage oligomeric matrix protein
| Autor: | Peiman Liu, Jacqueline T. Hecht, Karen L. Posey, Huiqiu R. Wang, Alka C. Veerisetty, Joseph L. Alcorn |
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| Rok vydání: | 2010 |
| Předmět: |
musculoskeletal diseases
Cellular pathology animal structures lcsh:Medicine Osteochondrodysplasias Chondrocyte Multiple epiphyseal dysplasia Rheumatology/Cartilage Biology and Osteoarthritis Small hairpin RNA Pseudoachondroplasia Mice fluids and secretions Chondrocytes medicine Extracellular Animals Matrilin Proteins RNA Messenger RNA Small Interfering lcsh:Science Glycoproteins Cartilage oligomeric matrix protein Extracellular Matrix Proteins Multidisciplinary biology Cartilage Genetics and Genomics/Gene Therapy lcsh:R Cell Biology/Extra-Cellular Matrix medicine.disease musculoskeletal system Molecular biology carbohydrates (lipids) Genetics and Genomics/Gene Function medicine.anatomical_structure Genetics and Genomics/Disease Models biology.protein lcsh:Q Mutant Proteins RNA Interference Rheumatology/Connective Tissue Disease Research Article |
| Zdroj: | PLoS ONE PLoS ONE, Vol 5, Iss 4, p e10302 (2010) |
| ISSN: | 1932-6203 |
| Popis: | Mutations in cartilage oligomeric matrix protein (COMP), a large extracellular glycoprotein expressed in musculoskeletal tissues, cause two skeletal dysplasias, pseudoachondroplasia and multiple epiphyseal dysplasia. These mutations lead to massive intracellular retention of COMP, chondrocyte death and loss of growth plate chondrocytes that are necessary for linear growth. In contrast, COMP null mice have only minor growth plate abnormalities, normal growth and longevity. This suggests that reducing mutant and wild-type COMP expression in chondrocytes may prevent the toxic cellular phenotype causing the skeletal dysplasias. We tested this hypothesis using RNA interference to reduce steady state levels of COMP mRNA. A panel of shRNAs directed against COMP was tested. One shRNA (3B) reduced endogenous and recombinant COMP mRNA dramatically, regardless of expression levels. The activity of the shRNA against COMP mRNA was maintained for up to 10 weeks. We also demonstrate that this treatment reduced ER stress. Moreover, we show that reducing steady state levels of COMP mRNA alleviates intracellular retention of other extracellular matrix proteins associated with the pseudoachondroplasia cellular pathology. These findings are a proof of principle and the foundation for the development of a therapeutic intervention based on reduction of COMP expression. |
| Databáze: | OpenAIRE |
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