title: Molecular Therapy for Degenerative Disc Disease: Clues from Secretome Analysis of the Notochordal Cell-Rich Nucleus Pulposus.
abstract: Degenerative disc disease (DDD) is associated with spinal pain often leading to long-term disability. However, the non-chondrodystrophic canine intervertebral disc is protected from the development of DDD, ostensibly due to its retention of notochordal cells (NC) in the nucleus pulposus (NP). In this study, we hypothesized that secretome analysis of the NC-rich NP will lead to the identification of key proteins that delay the onset of DDD. Using mass-spectrometry, we identified 303 proteins including components of TGFβ- and Wnt-signaling, anti-angiogeneic factors and proteins that inhibit axonal ingrowth in the bioactive fractions of serum free, notochordal cell derived conditioned medium (NCCM). Ingenuity Pathway Analysis revealed TGFβ1 and CTGF as major hubs in protein interaction networks. In vitro treatment with TGFβ1 and CTGF promoted the synthesis of healthy extra-cellular matrix proteins, increased cell proliferation and reduced cell death in human degenerative disc NP cells. A single intra-discal injection of recombinant TGFβ1 and CTGF proteins in a pre-clinical rat-tail disc injury model restored the NC and stem cell rich NP. In conclusion, we demonstrate the potential of TGFβ1 and CTGF to mitigate the progression of disc degeneration and the potential use of these molecules in a molecular therapy to treat the degenerative disc.
authors.affiliation: Krembil Research Institute, Toronto Western Hospital, Toronto, Ontario, Canada.
authors.forename: Ajay
authors.initials: A
authors.lastname: Matta
authors.affiliation: Krembil Research Institute, Toronto Western Hospital, Toronto, Ontario, Canada.
authors.forename: M Zia
authors.initials: MZ
authors.lastname: Karim
authors.affiliation: Department of Biochemistry, Faculty of Medicine, University of Toronto, Toronto, Ontario, Canada.
authors.forename: David E
authors.initials: DE
authors.lastname: Isenman
authors.affiliation: Krembil Research Institute, Toronto Western Hospital, Toronto, Ontario, Canada.|Department of Surgery, University of Toronto, Toronto, Ontario, Canada.|Division of Research, Canadian Memorial Chiropractic College, Canada.
authors.forename: W Mark
authors.initials: WM
authors.lastname: Erwin
mesh_terms: D000818:Animals; D055511:CCN Intercellular Signaling Proteins / Q000378:metabolism; D002470:Cell Survival; D002478:Cells, Cultured; D055513:Connective Tissue Growth Factor / Q000378:metabolism; D017077:Culture Media, Conditioned; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D055959:Intervertebral Disc Degeneration / Q000378:metabolism* / Q000473:pathology / Q000628:therapy*; D013058:Mass Spectrometry; D009672:Notochord / Q000378:metabolism*; D000070614:Nucleus Pulposus / Q000378:metabolism*; D060066:Protein Interaction Maps; D017208:Rats, Wistar; D012097:Repressor Proteins / Q000378:metabolism; D015398:Signal Transduction; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: D055511:CCN Intercellular Signaling Proteins; C526317:CCN2 protein, human; C413323:CCN5 protein, human; D017077:Culture Media, Conditioned; D012097:Repressor Proteins; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; D055513:Connective Tissue Growth Factor