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1. Asporin expression is highly regulated in human chondrocytes

Author

Duval, Elise, Bigot, Nicolas, Hervieu, Magalie, Kou, Ikuyo, Leclercq, Sylvain, Galéra, Philippe, Boumediene, Karim, and Baugé, Catherine

Subjects
Quantitative Biology - Tissues and Organs
Abstract

A significant association between a polymorphism in the D repeat of the gene encoding asporin and osteoarthritis, the most frequent of articular diseases, has been recently reported. The goal of the present study was to investigate the expression of this new class I small leucine-rich proteoglycan (SLRP) in human articular chondrocytes. First, we studied the modulation of asporin (ASPN) expression by cytokines by Western blot and reverse transcription-polymerase chain reaction. Interleukin-1$\beta$ and tumor necrosis factor-$\alpha$ downregulated ASPN, whereas transforming growth factor-$\beta$1 (when incubated in a serum-free medium) upregulated it. Similarly to proinflammatory cytokines, chondrocyte dedifferentiation induced by a successive passages of cells was accompanied by a decreased asporin expression, whereas their redifferentiation by three-dimensional culture restored its expression. Finally, we found an important role of the transcription factor Sp1 in the regulation of ASPN expression. Sp1 ectopic expression increased ASPN mRNA level and promoter activity. In addition, using gene reporter assay and electrophoretic mobility shift assay, we showed that Sp1 mediated its effect through a region located between -473 and -140 bp upstream of the transcription start site in ASPN gene. In conclusion, this report is the first study on the regulation of asporin expression by different cytokines in human articular chondrocytes. Our data indicate that the expression of this gene is finely regulated in cartilage and suggest a major role of Sp1.

Published
2015
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2. Characterization and use of Equine Bone Marrow Mesenchymal Stem Cells in Equine Cartilage Engineering. Study of their Hyaline Cartilage Forming Potential when Cultured under Hypoxia within a Biomaterial in the Presence of BMP-2 and TGF-ß1

Author

Branly, Thomas, Bertoni, Lélia, Contentin, Romain, Rakic, Rodolphe, Gomez-Leduc, Tangni, Desancé, Mélanie, Hervieu, Magalie, Legendre, Florence, Jacquet, Sandrine, Audigié, Fabrice, Denoix, Jean-Marie, Demoor, Magali, and Galéra, Philippe

Published
2017
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6. Enhanced chondrogenesis of bone marrow-derived stem cells by using a combinatory cell therapy strategy with BMP-2/TGF-β1, hypoxia, and COL1A1/HtrA1 siRNAs

Author

Legendre, Florence, Ollitrault, David, Gómez-Leduc, Tangni, Bouyoucef, Mouloud, Hervieu, Magalie, Gruchy, Nicolas, Mallein-Gerin, Frédéric, Leclercq, Sylvain, Demoor, Magali, Galéra, Philippe, Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Service de Génétique [CHU Caen], CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU), Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Science, Cell- and Tissue-Based Therapy, Bone Morphogenetic Protein 2, Mice, Nude, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Mesenchymal Stem Cell Transplantation, Collagen Type I, Article, Transforming Growth Factor beta1, Mice, Chondrocytes, Bone Marrow, Osteoarthritis, chondrogenesis, Animals, Humans, RNA, Small Interfering, Cells, Cultured, Aged, Aged, 80 and over, Tissue Engineering, hypoxia, Cell Differentiation, Mesenchymal Stem Cells, High-Temperature Requirement A Serine Peptidase 1, Middle Aged, BM-MSC, Collagen Type I, alpha 1 Chain, siRNA, Medicine, Female
Abstract

International audience; Mesenchymal stem cells (MSCs) hold promise for cartilage engineering. Here, we aimed to determine the best culture conditions to induce chondrogenesis of MSCs isolated from bone marrow (BM) of aged osteoarthritis (OA) patients. We showed that these BM-MSCs proliferate slowly, are not uniformly positive for stem cell markers, and maintain their multilineage potential throughout multiple passages. The chondrogenic lineage of BM-MSCs was induced in collagen scaffolds, under normoxia or hypoxia, by BMP-2 and/or TGF-β1. The best chondrogenic induction, with the least hypertrophic induction, was obtained with the combination of BMP-2 and TGF-β1 under hypoxia. Differentiated BM-MSCs were then transfected with siRNAs targeting two markers overexpressed in OA chondrocytes, type I collagen and/or HtrA1 protease. siRNAs significantly decreased mRNA and protein levels of type I collagen and HtrA1, resulting in a more typical chondrocyte phenotype, but with frequent calcification of the subcutaneously implanted constructs in a nude mouse model. Our 3D culture model with BMP-2/TGF-β1 and COL1A1/HtrA1 siRNAs was not effective in producing a cartilage-like matrix in vivo. Further optimization is needed to stabilize the chondrocyte phenotype of differentiated BM-MSCs. Nevertheless, this study offers the opportunity to develop a combinatory cellular therapy strategy for cartilage tissue engineering.

Published
2017
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7. Enhanced chondrogenesis of bone marrow-derived stem cells by using a combinatory cell therapy strategy with BMP-2/TGF-β1, hypoxia, and COL1A1/HtrA1 siRNAs

Author

Branly, Thomas, Contentin, Romain, Desancé, Mélanie, Jacquel, Thibaud, Bertoni, Lelia, JACQUET, Sandrine, Denoix, Jean-Marie, Audigié, Fabrice, Legendre, Florence, Ollitrault, David, Gómez-Leduc, Tangni, Bouyoucef, Mouloud, Hervieu, Magalie, Gruchy, Nicolas, Mallein-Gerin, Frédéric, Leclercq, Sylvain, Demoor, Magali, Galéra, Philippe, Microenvironnement cellulaire et pathologie (MILPAT), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Biomécanique et Pathologie Locomotrice du Cheval (BPLC), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Institut universitaire de formation des maîtres - Champagne-Ardenne (IUFM Champagne-Ardenne), Université de Reims Champagne-Ardenne (URCA), Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Institute of cardiometabolism and nutrition (ICAN), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [APHP]-Sorbonne Université (SU), Normandie Université (NU), Matrice extracellulaire normale et pathologique, Laboratoire de cytogénétique prénatale [CHU Caen], Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), EDF R&D (EDF R&D), EDF (EDF), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], and Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)

Subjects
[SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2017

8. Hypoxia Is a Critical Parameter for Chondrogenic Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells in Type I/III Collagen Sponges

Author

Gómez-Leduc, Tangni, Desancé, Mélanie, Hervieu, Magalie, Legendre, Florence, Ollitrault, David, de Vienne, Claire, Herlicoviez, Michel, Galéra, Philippe, Demoor, Magali, Microenvironnement cellulaire et pathologie (MILPAT), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), and Service de Gynécologie-Obstétrique et Médecine de la Reproduction, CHU Caen

Subjects
Cartilage, Articular, [SDV]Life Sciences [q-bio], Bone Morphogenetic Protein 2, Gene Expression, cartilage tissue engineering, Article, Collagen Type I, Transforming Growth Factor beta1, lcsh:Chemistry, Chondrocytes, chondrogenesis, Humans, Cell Lineage, lcsh:QH301-705.5, Cells, Cultured, ComputingMilieux_MISCELLANEOUS, mesenchymal stem cells, hypoxia, Cell Differentiation, Fetal Blood, Extracellular Matrix, Oxygen, Collagen Type III, Phenotype, lcsh:Biology (General), lcsh:QD1-999, umbilical cord blood, Biomarkers
Abstract

Umbilical cord blood (UCB) is an attractive alternative to bone marrow for isolation of mesenchymal stem cells (MSCs) to treat articular cartilage defects. Here, we set out to determine the growth factors (bone morphogenetic protein 2 (BMP-2) and transforming growth factor-β (TGF-β1)) and oxygen tension effects during chondrogenesis of human UCB-MSCs for cartilage engineering. Chondrogenic differentiation was induced using 3D cultures in type I/III collagen sponges with chondrogenic factors in normoxia (21% O2) or hypoxia (

Published
2017
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9. RNA Interference and BMP-2 Stimulation Allows Equine Chondrocytes Redifferentiation in 3D-Hypoxia Cell Culture Model: Application for Matrix-Induced Autologous Chondrocyte Implantation

Author

Rakic, Rodolphe, Bourdon, Bastien, Hervieu, Magalie, Branly, Thomas, Legendre, Florence, Saulnier, Nathalie, Audigié, Fabrice, Maddens, Stéphane, Demoor, Magali, Galéra, Philippe, Biologie, génétique et thérapies ostéoarticulaires et respiratoires (BIOTARGEN), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Microenvironnement cellulaire et pathologie (MILPAT), Vetbiobank, Biomécanique et Pathologie Locomotrice du Cheval (BPLC), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), EFS Rhône-Alpes, and EFS

Subjects
Cartilage, Articular, [SDV]Life Sciences [q-bio], type II collagen, Cell Culture Techniques, Bone Morphogenetic Protein 2, Article, Collagen Type I, lcsh:Chemistry, Chondrocytes, BMP-2, Animals, matrix-associated autologous chondrocyte implantation (MACI), Horses, RNA, Small Interfering, lcsh:QH301-705.5, ComputingMilieux_MISCELLANEOUS, hypoxia, dedifferentiation, Cell Differentiation, equine model, Cell Hypoxia, Extracellular Matrix, Col1a1, Collagen Type III, Phenotype, lcsh:Biology (General), lcsh:QD1-999, tissue engineering, siRNA, chondrocyte, HtrA1, RNA Interference, Biomarkers
Abstract

International audience; As in humans, osteoarthritis (OA) causes considerable economic loss to the equine industry. New hopes for cartilage repair have emerged with the matrix-associated autologous chondrocyte implantation (MACI). Nevertheless, its limitation is due to the dedifferentiation occurring during the chondrocyte amplification phase, leading to the loss of its capacity to produce a hyaline extracellular matrix (ECM). To enhance the MACI therapy efficiency, we have developed a strategy for chondrocyte redifferentiation, and demonstrated its feasibility in the equine model. Thus, to mimic the cartilage microenvironment, the equine dedifferentiated chondrocytes were cultured in type I/III collagen sponges for 7 days under hypoxia in the presence of BMP-2. In addition, chondrocytes were transfected by siRNA targeting Col1a1 and Htra1 mRNAs, which are overexpressed during dedifferentiation and OA. To investigate the quality of the neo-synthesized ECM, specific and atypical cartilage markers were evaluated by RT-qPCR and Western blot. Our results show that the combination of 3D hypoxia cell culture, BMP-2 (Bone morphogenetic protein-2), and RNA interference, increases the chondrocytes functional indexes (Col2a1/Col1a1, Acan/Col1a1), leading to an effective chondrocyte redifferentiation. These data represent a proof of concept for this process of application, in vitro, in the equine model, and will lead to the improvement of the MACI efficiency for cartilage tissue engineering therapy in preclinical/clinical trials, both in equine and human medicine.

Published
2017
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11. RNA Interference and BMP-2 Stimulation Allows Equine Chondrocytes Redifferentiation in 3D-Hypoxia Cell Culture Model: Application for Matrix-Induced Autologous Chondrocyte Implantation.

Author

Bourdon, Bastien, Hervieu, Magalie, Branly, Thomas, Legendre, Florence, Demoor, Magali, Galera, Philippe, Rakic, Rodolphe, Saulnier, Nathalie, Maddens, Stéphane, and Audigié, Fabrice

Subjects
*RNA interference, *BONE morphogenetic proteins, *CARTILAGE cells, *OSTEOARTHRITIS, *HORSE diseases, *HYALINE membrane disease
Abstract

As in humans, osteoarthritis (OA) causes considerable economic loss to the equine industry. New hopes for cartilage repair have emerged with the matrix-associated autologous chondrocyte implantation (MACI). Nevertheless, its limitation is due to the dedifferentiation occurring during the chondrocyte amplification phase, leading to the loss of its capacity to produce a hyaline extracellular matrix (ECM). To enhance the MACI therapy efficiency, we have developed a strategy for chondrocyte redifferentiation, and demonstrated its feasibility in the equine model. Thus, to mimic the cartilage microenvironment, the equine dedifferentiated chondrocytes were cultured in type I/III collagen sponges for 7 days under hypoxia in the presence of BMP-2. In addition, chondrocytes were transfected by siRNA targeting Col1a1 and Htra1 mRNAs, which are overexpressed during dedifferentiation and OA. To investigate the quality of the neo-synthesized ECM, specific and atypical cartilage markers were evaluated by RT-qPCR and Western blot. Our results show that the combination of 3D hypoxia cell culture, BMP-2 (Bone morphogenetic protein-2), and RNA interference, increases the chondrocytes functional indexes (Col2a1/Col1a1, Acan/Col1a1), leading to an effective chondrocyte redifferentiation. These data represent a proof of concept for this process of application, in vitro, in the equine model, and will lead to the improvement of the MACI efficiency for cartilage tissue engineering therapy in preclinical/clinical trials, both in equine and human medicine. [ABSTRACT FROM AUTHOR]

Published
2017
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12. Enhanced Hyaline Cartilage Matrix Synthesis in Collagen Sponge Scaffolds by Using siRNA to Stabilize Chondrocytes Phenotype Cultured with Bone Morphogenetic Protein-2 Under Hypoxia

Author

Legendre, Florence, primary, Ollitrault, David, additional, Hervieu, Magalie, additional, Baugé, Catherine, additional, Maneix, Laure, additional, Goux, Didier, additional, Chajra, Hanane, additional, Mallein-Gerin, Frédéric, additional, Boumediene, Karim, additional, Galera, Philippe, additional, and Demoor, Magali, additional

Published
2013
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15. PARP-1 Inhibition Increases Oxidative Stress in Ets-1-Expressing MDA-MB-231 Breast Cancer Cells.

Author

Hervieu M, Legrand AJ, Floquet E, Idziorek T, Spriet C, Monté D, Villeret V, Aumercier M, and Choul-Li S

Subjects
Humans, Female, Cell Line, Tumor, Reactive Oxygen Species metabolism, Cell Proliferation drug effects, Gene Expression Regulation, Neoplastic drug effects, DNA Damage drug effects, Phenanthrenes, Oxidative Stress drug effects, Breast Neoplasms pathology, Breast Neoplasms drug therapy, Breast Neoplasms metabolism, Breast Neoplasms genetics, Poly (ADP-Ribose) Polymerase-1 metabolism, Poly (ADP-Ribose) Polymerase-1 antagonists & inhibitors, Poly(ADP-ribose) Polymerase Inhibitors pharmacology, Proto-Oncogene Protein c-ets-1 metabolism, Proto-Oncogene Protein c-ets-1 genetics
Abstract

Background: The Ets-1 transcription factor plays a primordial role in regulating the expression of numerous genes implicated in cancer progression. In a previous study, we revealed that poly(ADP-ribose) polymerase-1 (PARP-1) inhibition by PJ-34 results in Ets-1 level increase in cells, which is related with cell death of Ets-1-expressing cancer cells., Aims: The mechanism of the antitumor effect of PARP-1 inhibition was investigated in the Ets-1-expressing MDA-MB-231 breast cancer cells., Methods and Results: We tested the effects of four PARP inhibitors (PARPi) (PJ-34, Veliparib, Olaparib, and Rucaparib). We first demonstrated that PARPi reduced cells growth through G2/M cell cycle arrest. Next, we evaluated PARP-1 inhibition effect on oxidative DNA damage in Ets-1-overexpressing and Ets-1-non-expressing breast cancer cells and we showed that PARPi led only Ets-1-overexpressing cells to accumulate it, which triggers the DNA damage response as revealed by the increase in the level of a panel of DNA damage-related proteins. Importantly, we demonstrated that PARPi increased reactive oxygen species (ROS), only in Ets-1-overexpressing cells and this is accompanied by upregulation of p47 phox expression, a subunit of the NAPDH oxidase (NOX)., Conclusion: These preliminary findings correlate PARPi-induced oxidative DNA damage/oxidative stress to Ets-1 expression in breast cancer cells., (© 2025 The Author(s). Cancer Reports published by Wiley Periodicals LLC.)

Published
2025
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