title: rAAV-mediated overexpression of sox9, TGF-β and IGF-I in minipig bone marrow aspirates to enhance the chondrogenic processes for cartilage repair.
abstract: Administration of therapeutic gene sequences coding for chondrogenic and chondroreparative factors in bone marrow aspirates using the clinically adapted recombinant adeno-associated virus (rAAV) vector may provide convenient, single-step approaches to improve cartilage repair. Here, we tested the ability of distinct rAAV constructs coding for the potent SOX9, transforming growth factor beta (TGF-β) and insulin-like growth factor I (IGF-I) candidate factors to modify marrow aspirates from minipigs to offer a preclinical large animal model system adapted for a translational evaluation of cartilage repair upon transplantation in sites of injury. Our results demonstrate that high, prolonged rAAV gene transfer efficiencies were achieved in the aspirates (up to 100% for at least 21 days) allowing to produce elevated amounts of the transcription factor SOX9 that led to increased levels of matrix synthesis and chondrogenic differentiation and of the growth factors TGF-β and IGF-I that both increased cell proliferation, matrix synthesis and chondrogenic differentiation (although to a lower level than SOX9) compared with control (lacZ) condition. Remarkably, application of the candidate SOX9 vector also led to reduced levels of hypertrophic differentiation in the aspirates, possibly by modulating the β-catenin, Indian hedgehog and PTHrP pathways. The present findings show the benefits of modifying minipig marrow concentrates via rAAV gene transfer as a future means to develop practical strategies to promote cartilage repair in a large animal model.
authors.affiliation: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Frisch
authors.affiliation: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Rey-Rico
authors.affiliation: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany.
authors.forename: J K
authors.initials: JK
authors.lastname: Venkatesan
authors.affiliation: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Schmitt
authors.affiliation: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany.|Department of Orthopaedic Surgery, Saarland University Medical Center, Homburg/Saar, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Madry
authors.affiliation: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Cucchiarini
mesh_terms: D000818:Animals; D016026:Bone Marrow Transplantation; D002356:Cartilage / Q000293:injuries; D020219:Chondrogenesis; D000229:Dependovirus / Q000235:genetics*; D005822:Genetic Vectors / Q000627:therapeutic use*; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000627:therapeutic use*; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000627:therapeutic use*; D013552:Swine; D013556:Swine, Miniature; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000627:therapeutic use*
chemical_list: D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D016212:Transforming Growth Factor beta; D007334:Insulin-Like Growth Factor I