Molecular Therapy for Degenerative Disc Disease: Clues from Secretome Analysis of the Notochordal CellRich Nucleus Pulposus

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 TGF beta-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 TGF beta 1 and CTGF as major hubs in protein interaction networks. In vitro treatment with TGF beta 1 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 intradiscal injection of recombinant TGF beta 1 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 TGF beta 1 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.