title: TGF-β1 and GDF5 Act Synergistically to Drive the Differentiation of Human Adipose Stromal Cells toward Nucleus Pulposus-like Cells.
abstract: Degenerative disc disease (DDD) primarily affects the central part of the intervertebral disc namely the nucleus pulposus (NP). DDD explains about 40% of low back pain and is characterized by massive cellular alterations that ultimately result in the disappearance of resident NP cells. Thus, repopulating the NP with regenerative cells is a promising therapeutic approach and remains a great challenge. The objectives of this study were to evaluate the potential of growth factor-driven protocols to commit human adipose stromal cells (hASCs) toward NP-like cell phenotype and the involvement of Smad proteins in this differentiation process. Here, we demonstrate that the transforming growth factor-β1 and the growth differentiation factor 5 synergistically drive the nucleopulpogenic differentiation process. The commitment of the hASCs was robust and highly specific as attested by the expression of NP-related genes characteristic of young healthy human NP cells. In addition, the engineered NP-like cells secreted an abundant aggrecan and type II collagen rich extracellular matrix comparable with that of native NP. Furthermore, we demonstrate that these in vitro engineered cells survived, maintained their specialized phenotype and secretory activity after in vivo transplantation in nude mice subcutis. Finally, we provide evidence suggesting that the Smad 2/3 pathway mainly governed the acquisition of the NP cell molecular identity while the Smad1/5/8 pathway controlled the NP cell morphology. This study offers valuable insights for the development of biologically-inspired treatments for DDD by generating adapted and exhaustively characterized autologous regenerative cells.
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.
authors.forename: Pauline
authors.initials: P
authors.lastname: Colombier
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.|Université de Nantes, UFR Sciences Biologiques et Pharmaceutiques, Nantes, France.|CHU Nantes, Pharmacie Centrale, PHU 11, Nantes, France.
authors.forename: Johann
authors.initials: J
authors.lastname: Clouet
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Boyer
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.
authors.forename: Maëva
authors.initials: M
authors.lastname: Ruel
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Bonin
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Lesoeur
authors.affiliation: Université de Nantes, UFR Médecine, Nantes, France.|CHU Nantes, Service d'Anatomopathologie, Nantes, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Moreau
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.|CRIP, Centre de Recherche et d'Investigations Précliniques, ONIRIS, Nantes, France.
authors.forename: Borhane-Hakim
authors.initials: BH
authors.lastname: Fellah
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.|CHU Nantes, PHU 4 OTONN, Nantes, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Weiss
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.|Université de Nantes, UFR Sciences et Techniques, Nantes, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Lescaudron
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Camus
authors.affiliation: INSERM UMRS 791, Laboratoire d'Ingénierie Osteo Articulaire et Dentaire (LIOAD), Nantes, France.|Université de Nantes, UFR Odontologie, Nantes, France.|CHU Nantes, PHU 4 OTONN, Nantes, France.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Guicheux
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000637:transplantation; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D060846:Cell Engineering / Q000379:methods; D005109:Extracellular Matrix; D055428:Growth Differentiation Factor 5 / Q000235:genetics* / Q000627:therapeutic use; D006801:Humans; D055959:Intervertebral Disc Degeneration / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D017116:Low Back Pain; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D051379:Mice; D000070614:Nucleus Pulposus / Q000166:cytology / Q000637:transplantation; D051785:Smad Proteins / Q000235:genetics; D053773:Transforming Growth Factor beta1 / Q000235:genetics* / Q000627:therapeutic use
chemical_list: C525893:GDF5 protein, human; D055428:Growth Differentiation Factor 5; D051785:Smad Proteins; D053773:Transforming Growth Factor beta1
keywords: Intervertebral disc; Regenerative medicine; Smad-driven nucleopulpogenic differentiation; Stem cells