title: Co-overexpression of TGF-β and SOX9 via rAAV gene transfer modulates the metabolic and chondrogenic activities of human bone marrow-derived mesenchymal stem cells.
abstract: BACKGROUND
  Articular cartilage has a limited potential for self-healing. Transplantation of genetically modified progenitor cells like bone marrow-derived mesenchymal stem cells (MSCs) is an attractive strategy to improve the intrinsic repair capacities of damaged articular cartilage.
  
  
  METHODS
  In this study, we examined the potential benefits of co-overexpressing the pleiotropic transformation growth factor beta (TGF-β) with the cartilage-specific transcription factor SOX9 via gene transfer with recombinant adeno-associated virus (rAAV) vectors upon the biological activities of human MSCs (hMSCs). Freshly isolated hMSCs were transduced over time with separate rAAV vectors carrying either TGF-β or sox9 in chondrogenically-induced aggregate cultures to evaluate the efficacy and duration of transgene expression and to monitor the effects of rAAV-mediated genetic modification upon the cellular activities (proliferation, matrix synthesis) and chondrogenic differentiation potency compared with control conditions (lacZ treatment, sequential transductions).
  
  
  RESULTS
  Significant, prolonged TGF-β/sox9 co-overexpression was achieved in chondrogenically-induced hMSCs upon co-transduction via rAAV for up to 21 days, leading to enhanced proliferative, biosynthetic, and chondrogenic activities relative to control treatments, especially when co-applying the candidate vectors at the highest vector doses tested. Optimal co-administration of TGF-β with sox9 also advantageously reduced hypertrophic differentiation of the cells in the conditions applied here.
  
  
  CONCLUSIONS
  The present findings demonstrate the possibility of modifying MSCs by combined therapeutic gene transfer as potent future strategies for implantation in clinically relevant animal models of cartilage defects in vivo.
authors.affiliation: Institute of Arthritis, Peking University People's Hospital, Beijing, 100044, P.R. China. taokepku@gmail.com.|Peking University Health Science Center, Beijing, 100191, P.R. China. taokepku@gmail.com.|Center of Experimental Orthopedics, Saarland University Medical Center, Kirrbergerstraße Bldg 37, Homburg/Saar, D-66421, Germany. taokepku@gmail.com.
authors.forename: Ke
authors.initials: K
authors.lastname: Tao
authors.affiliation: Center of Experimental Orthopedics, Saarland University Medical Center, Kirrbergerstraße Bldg 37, Homburg/Saar, D-66421, Germany. janina.frisch@gmx.de.
authors.forename: Janina
authors.initials: J
authors.lastname: Frisch
authors.affiliation: Center of Experimental Orthopedics, Saarland University Medical Center, Kirrbergerstraße Bldg 37, Homburg/Saar, D-66421, Germany. ana.rey.rico@gmail.com.
authors.forename: Ana
authors.initials: A
authors.lastname: Rey-Rico
authors.affiliation: Center of Experimental Orthopedics, Saarland University Medical Center, Kirrbergerstraße Bldg 37, Homburg/Saar, D-66421, Germany. jegadish.venki@gmail.com.
authors.forename: Jagadeesh K
authors.initials: JK
authors.lastname: Venkatesan
authors.affiliation: Center of Experimental Orthopedics, Saarland University Medical Center, Kirrbergerstraße Bldg 37, Homburg/Saar, D-66421, Germany. gertrud.schmitt@uniklinikum-saarland.de.
authors.forename: Gertrud
authors.initials: G
authors.lastname: Schmitt
authors.affiliation: Center of Experimental Orthopedics, Saarland University Medical Center, Kirrbergerstraße Bldg 37, Homburg/Saar, D-66421, Germany. henning.madry@uks.eu.|Department of Orthopaedic Surgery, Saarland University Medical Center, Kirrbergerstr. Bldg 37, Homburg/Saar, D-66421, Germany. henning.madry@uks.eu.
authors.forename: Henning
authors.initials: H
authors.lastname: Madry
authors.affiliation: Institute of Arthritis, Peking University People's Hospital, Beijing, 100044, P.R. China. jianhao_lin@yahoo.com.|Peking University Health Science Center, Beijing, 100191, P.R. China. jianhao_lin@yahoo.com.
authors.forename: Jianhao
authors.initials: J
authors.lastname: Lin
authors.affiliation: Center of Experimental Orthopedics, Saarland University Medical Center, Kirrbergerstraße Bldg 37, Homburg/Saar, D-66421, Germany. mmcucchiarini@hotmail.com.
authors.forename: Magali
authors.initials: M
authors.lastname: Cucchiarini
mesh_terms: D000368:Aged; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D020219:Chondrogenesis; D000229:Dependovirus / Q000235:genetics; D005260:Female; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D055758:SOX9 Transcription Factor / Q000096:biosynthesis* / Q000235:genetics; D014161:Transduction, Genetic; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D016212:Transforming Growth Factor beta
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