Stimulation of proteoglycan synthesis by glucuronosyltransferase-1 gene delivery: a strategy to promote cartilage repair

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 -