Abstract
Objective
To investigate the role of endogenous TSG-6 in human osteoarthritis (OA) and assess the disease-modifying potential of a TSG-6-based biological treatment in cell, explant and animal models of OA.
Design
Knee articular cartilages from OA patients were analyzed for TSG-6 protein and mRNA expression using immunohistochemistry and RNAscope, respectively. The inhibitory activities of TSG-6 and its isolated Link module (Link_TSG6) on cytokine-induced degradation of OA cartilage explants were compared. Human mesenchymal stem/stromal cell-derived chondrocyte pellet cultures were used to determine the effects of Link_TSG6 and full-length TSG-6 on IL-1α-, IL-1β-, or TNF-stimulated ADAMTS4, ADAMTS5, and MMP13 mRNA expression. Link_TSG6 was administered i.a. to the rat ACLTpMMx model; cartilage damage and tactile allodynia were assessed.
Results
TSG-6 is predominantly associated with chondrocytes in regions of cartilage damage where high TSG-6 expression aligns with low MMP13, the major collagenase implicated in OA progression. Link_TSG6 is more potent than full-length TSG-6 at inhibiting cytokine-mediated matrix breakdown in human OA cartilage explants;>50% of donor cartilages, from 59 tested, were responsive to Link_TSG6 treatment. Link_TSG6 also displayed more potent effects in 3D pellet cultures, suppressing ADAMTS4, ADAMTS5, and MMP13 gene expression, which was consistent with reduced aggrecanase and collagenase activities in explant cultures. Link_TSG6 treatment reduced touch-evoked pain behavior and dose-dependently inhibited cartilage damage in a rodent model of surgically-induced OA.
Conclusions
Link_TSG6 has enhanced chondroprotective activity compared to the full-length TSG-6 protein and shows potential as a disease modifying OA drug via its inhibition of aggrecanase and collagenase activity.
Abbreviations
ACLTpMMxAnterior Cruciate Ligament Transection with partial Medial MeniscectomyADAMTSA Disintegrin and Metalloproteinase with Thrombospondin motifsAMGAnteromedial GonarthrosisANCOVAAnalysis of CovarianceANOVAAnalysis of VarianceBMIBody Mass IndexCTXIIC-terminal Cross-linked Telopeptide of Type II CollagenCXCL8C-X-C motif Chemokine Ligand 8DMBdimethylmethylene blueDMOADDisease Modifying Osteoarthritis DrugELISAEnzyme-linked Immunosorbent AssayGAGglycosaminoglycanHAhyaluronanhMSChuman Mesenchymal Stem/Stromal Celli.a.intra articularIL-1Interleukin-1JNKc-Jun N-terminal KinaseLink_TSG6the Link module of human TSG-6MMPMatrix MetalloproteinasemRNAmessenger Ribonucleic AcidNF-kBNuclear Factor kappa-light-chain-enhancer of activated B cellsOAosteoarthritisOSMOncostatin MqPCRquantitative Polymerase Chain ReactionRArheumatoid arthritisrhTSG-6recombinant human Tumour Necrosis Factor-Stimulated Gene-6TKAtotal knee arthroplastyTNAIP6Tumour Necrosis Factor-Induced Protein 6TNFTumour Necrosis FactorTSG-6Tumour Necrosis Factor-Stimulated Gene-6
To investigate the role of endogenous TSG-6 in human osteoarthritis (OA) and assess the disease-modifying potential of a TSG-6-based biological treatment in cell, explant and animal models of OA.
Design
Knee articular cartilages from OA patients were analyzed for TSG-6 protein and mRNA expression using immunohistochemistry and RNAscope, respectively. The inhibitory activities of TSG-6 and its isolated Link module (Link_TSG6) on cytokine-induced degradation of OA cartilage explants were compared. Human mesenchymal stem/stromal cell-derived chondrocyte pellet cultures were used to determine the effects of Link_TSG6 and full-length TSG-6 on IL-1α-, IL-1β-, or TNF-stimulated ADAMTS4, ADAMTS5, and MMP13 mRNA expression. Link_TSG6 was administered i.a. to the rat ACLTpMMx model; cartilage damage and tactile allodynia were assessed.
Results
TSG-6 is predominantly associated with chondrocytes in regions of cartilage damage where high TSG-6 expression aligns with low MMP13, the major collagenase implicated in OA progression. Link_TSG6 is more potent than full-length TSG-6 at inhibiting cytokine-mediated matrix breakdown in human OA cartilage explants;>50% of donor cartilages, from 59 tested, were responsive to Link_TSG6 treatment. Link_TSG6 also displayed more potent effects in 3D pellet cultures, suppressing ADAMTS4, ADAMTS5, and MMP13 gene expression, which was consistent with reduced aggrecanase and collagenase activities in explant cultures. Link_TSG6 treatment reduced touch-evoked pain behavior and dose-dependently inhibited cartilage damage in a rodent model of surgically-induced OA.
Conclusions
Link_TSG6 has enhanced chondroprotective activity compared to the full-length TSG-6 protein and shows potential as a disease modifying OA drug via its inhibition of aggrecanase and collagenase activity.
Abbreviations
ACLTpMMxAnterior Cruciate Ligament Transection with partial Medial MeniscectomyADAMTSA Disintegrin and Metalloproteinase with Thrombospondin motifsAMGAnteromedial GonarthrosisANCOVAAnalysis of CovarianceANOVAAnalysis of VarianceBMIBody Mass IndexCTXIIC-terminal Cross-linked Telopeptide of Type II CollagenCXCL8C-X-C motif Chemokine Ligand 8DMBdimethylmethylene blueDMOADDisease Modifying Osteoarthritis DrugELISAEnzyme-linked Immunosorbent AssayGAGglycosaminoglycanHAhyaluronanhMSChuman Mesenchymal Stem/Stromal Celli.a.intra articularIL-1Interleukin-1JNKc-Jun N-terminal KinaseLink_TSG6the Link module of human TSG-6MMPMatrix MetalloproteinasemRNAmessenger Ribonucleic AcidNF-kBNuclear Factor kappa-light-chain-enhancer of activated B cellsOAosteoarthritisOSMOncostatin MqPCRquantitative Polymerase Chain ReactionRArheumatoid arthritisrhTSG-6recombinant human Tumour Necrosis Factor-Stimulated Gene-6TKAtotal knee arthroplastyTNAIP6Tumour Necrosis Factor-Induced Protein 6TNFTumour Necrosis FactorTSG-6Tumour Necrosis Factor-Stimulated Gene-6
| Original language | English |
|---|---|
| Journal | Osteoarthritis Cartilage |
| Early online date | 29 May 2023 |
| DOIs | |
| Publication status | E-pub ahead of print - 29 May 2023 |
Keywords
- TSG-6
- Link_TSG6
- Osteoarthritis
- Chondrocyte
- Inflammation
- OA models
