TY - JOUR
T1 - Stimulation of proteoglycan synthesis by glucuronosyltransferase-1 gene delivery: a strategy to promote cartilage repair
AU - Venkatesan, N.
AU - Barre, L.
AU - Benani, A.
AU - Netter, Patrick
AU - Magdalou, Jacques
AU - Fournel-Gigleux, Sylvie
AU - Ouzzine, Mohamed
N1 -
dc.publisher: National Academy of Sciences
dc.description.sponsorship: Fonds National pour la Science (Actions Concertées Incitatives)
IT2B program
Institut National de la Santé et de la Recherche Médicale (Programme National de Recherches sur les Maladies Ostéo-Articulaire)
Ligue Régionale contre le Cancer
Contrat de Programme de Recherche Clinique
Programme Hospitalier de Recherche Clinique Centre Hospitalier Universitaire
PY - 2004/12
Y1 - 2004/12
N2 - Osteoarthritis is a degenerative joint disease characterized by a progressive loss of articular cartilage components, mainly proteoglycans (PGs), leading to destruction of the tissue. We investigate a therapeutic strategy based on stimulation of PG synthesis by gene transfer of the glycosaminoglycan (GAG)-synthesizing enzyme, ß1,3-glucuronosyltransferase-I (GlcAT-I) to promote cartilage repair. We previously reported that IL-1ß down-regulated the expression and activity of GlcAT-I in primary rat chondrocytes. Here, by using antisense oligonucleotides, we demonstrate that GlcAT-I inhibition impaired PG synthesis and deposition in articular cartilage explants, emphasizing the crucial role of this enzyme in PG anabolism. Thus, primary chondrocytes and cartilage explants were engineered by lipid-mediated gene delivery to efficiently overexpress a human GlcAT-I cDNA. Interestingly, GlcAT-I overexpression significantly enhanced GAG synthesis and deposition as evidenced by 35S-sulfate incorporation, histology, estimation of GAG content, and fluorophore-assisted carbohydrate electrophoresis analysis. Metabolic labeling and Western blot analyses further suggested that GlcAT-I expression led to an increase in the abundance rather than in the length of GAG chains. Importantly, GlcAT-I delivery was able to overcome IL-1ß-induced PG depletion and maintain the anabolic activity of chondrocytes. Moreover, GlcAT-I also restored PG synthesis to a normal level in cartilage explants previously depleted from endogenous PGs by IL-1ß-treatment. In concert, our investigations strongly indicated that GlcAT-I was able to control and reverse articular cartilage defects in terms of PG anabolism and GAG content associated with IL-1ß. This study provides a basis for a gene therapy approach to promote cartilage repair in degenerative joint diseases.
AB - Osteoarthritis is a degenerative joint disease characterized by a progressive loss of articular cartilage components, mainly proteoglycans (PGs), leading to destruction of the tissue. We investigate a therapeutic strategy based on stimulation of PG synthesis by gene transfer of the glycosaminoglycan (GAG)-synthesizing enzyme, ß1,3-glucuronosyltransferase-I (GlcAT-I) to promote cartilage repair. We previously reported that IL-1ß down-regulated the expression and activity of GlcAT-I in primary rat chondrocytes. Here, by using antisense oligonucleotides, we demonstrate that GlcAT-I inhibition impaired PG synthesis and deposition in articular cartilage explants, emphasizing the crucial role of this enzyme in PG anabolism. Thus, primary chondrocytes and cartilage explants were engineered by lipid-mediated gene delivery to efficiently overexpress a human GlcAT-I cDNA. Interestingly, GlcAT-I overexpression significantly enhanced GAG synthesis and deposition as evidenced by 35S-sulfate incorporation, histology, estimation of GAG content, and fluorophore-assisted carbohydrate electrophoresis analysis. Metabolic labeling and Western blot analyses further suggested that GlcAT-I expression led to an increase in the abundance rather than in the length of GAG chains. Importantly, GlcAT-I delivery was able to overcome IL-1ß-induced PG depletion and maintain the anabolic activity of chondrocytes. Moreover, GlcAT-I also restored PG synthesis to a normal level in cartilage explants previously depleted from endogenous PGs by IL-1ß-treatment. In concert, our investigations strongly indicated that GlcAT-I was able to control and reverse articular cartilage defects in terms of PG anabolism and GAG content associated with IL-1ß. This study provides a basis for a gene therapy approach to promote cartilage repair in degenerative joint diseases.
KW - Osteoarthritis
KW - Gene transfer
KW - Chondrocyte
KW - Glycosyltransferase
KW - Glycosaminoglycans
U2 - 10.1073/pnas.0404504102
DO - 10.1073/pnas.0404504102
M3 - Article
C2 - 15601778
SN - 0027-8424
VL - 101
SP - 18087
EP - 18092
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 52
ER -