Intervertebral disc cells from human back pain subjects exhibit TNFR1-mediated senescence and lack TNFR2-modulated repair capacity

Gansau, J.; Grossi, E.; Rodriguez, L.; Wang, M.; Laudier, D.; Chaudhary, S.; Hecht, A.; Fu, W.; Sebra, R.; Liu, C.; Iatridis, J. C.

doi:10.1101/2024.02.22.581620

bioRxiv · 10.1101/2024.02.22.581620

Intervertebral disc cells from human back pain subjects exhibit TNFR1-mediated senescence and lack TNFR2-modulated repair capacity

Gansau, J.·Grossi, E.·Rodriguez, L.·Wang, M.·Laudier, D.·Chaudhary, S.·Hecht, A.·Fu, W.·Sebra, R.·Liu, C.·Iatridis, J. C.

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Abstract

Poor intervertebral disc (IVD) healing causes IVD degeneration (IVDD) and progression to herniation and back pain. This study identified distinct roles of TNF-receptors (TNFRs) in contributing to poor healing in painful IVDD. We first isolated IVDD tissue of back pain subjects and determined the complex pro-inflammatory mixture contained many chemokines for recruiting inflammatory cells. Single-cell RNA-sequencing of human IVDD tissues revealed these pro- inflammatory cytokines were dominantly expressed by a small macrophage-population. Human annulus fibrosus (hAF) cells treated with IVDD-conditioned media (CM) underwent senescence with greatly reduced metabolic rates and limited inflammatory responses. TNFR1 inhibition partially restored hAF cell metabolism sufficiently to enable a robust chemokine and cytokine response to CM. We showed that the pro-reparative TNFR2 was very limited on hIVD cell membranes so that TNFR2 inhibition with blocking antibodies or activation using Atsttrin had no effect on hAF cells with CM challenge. However, TNFR2 was expressed in high levels on macrophages identified in scRNA-seq analyses, suggesting their role in repair responses. Results therefore point to therapeutic strategies for painful IVDD involving immunomodulation of TNFR1 signaling in IVD cells to enhance metabolism and enable a more robust inflammatory response including recruitment or delivery of TNFR2 expressing immune cells to enhance IVD repair. SUMMARY STATEMENTTNFR1 signaling drives cells towards senesce and muted inflammatory response in painful intervertebral disc degeneration, while limited TNFR2 signaling may limit disc cell repair responses.

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Gansau, J., Grossi, E., Rodriguez, L., Wang, M., Laudier, D., Chaudhary, S., Hecht, A., Fu, W., Sebra, R., Liu, C., Iatridis, J. C.. 2024-02-25. Intervertebral disc cells from human back pain subjects exhibit TNFR1-mediated senescence and lack TNFR2-modulated repair capacity. https://doi.org/10.1101/2024.02.22.581620

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