Role of the terminal complement complex in intervertebral disc degeneration
Final Report Abstract
The project focused on basic investigations concerning the implication and role of the terminal complement complex (TCC) in disc degeneration (DD). Immunohistochemical analyses in a large patient cohort clearly indicated that TCC deposition is associated with the disease progression and its presence positively correlated with the degree of disc degeneration assessed by the MRI-based Pfirrmann grading. Furthermore, IgG and cathepsin D, which represent triggers of complement activation, were identified in annulus fibrosus (AF), nucleus pulposus (NP) as well as endplate (EP) tissue from DD patients. Subsequent in vitro studies confirmed that cells in human disc tissue and isolated human disc cells cannot protect themselves completely from TCC deposition under exposition to human serum, although the expression of CD46, CD55 and CD59 increased during in vitro culturing. Proinflammatory cytokines like IL-1β and TNF-α, which are known to be involved in the pathogenetic process of DD did not further increase terminal complement activation. In contrast, IL-1β revealed a certain protective influence on TCC deposition in tissue culture. This represents an interesting observation which deserves further investigation. The possibility of direct C5 cleavage in the presence of AF-cells could be shown in a serum-free assay in which the generation of C5a from added C5 was tested. Besides cathepsin D also endplate tissue-conditioned medium from patients with DD could induce this direct activation process of the terminal complement cascade also leading to the generation of C5b as the starting component of TCC formation. To address the question of a functional relevance of terminal complement activation an AF-cell culture model with zymosan and 10% human serum was established which led to robust complement activation associated with anaphylatoxin generation and TCC deposition. Inhibition of terminal complement activation by Eculizumab indicated significant downregulation of several genes which are relevant in DD pathophysiology such as MMP1, ADAMTS4 and COX2. This represents the first evidence of a biological impact of terminal complement activation in human disc cells. Finally, a C6-deficient rabbit model based on a recessive mutation of the C6-gene which is characterized by the inability to form a functional TCC was used to study a potential implication in injury-induced disc degeneration in vivo. Therefore, two types of disc injury were tested: a minor trauma model using a needle puncture with 20 G and a new endplate drilling model (drill-diameter of 1.5 mm) which was developed within the project. In heterozygous C6-sufficient (C6+/-) and homozygous C6-deficient (C6-/-) rabbits the needle puncture did not lead to significant consequences in MRI T2-values, Pfirrmann Score and disc height index (DHI) 12 weeks after injury. In contrast, the drilling model led to a significantly increased Pfirrmann score, and a tendency for lower T2 MRI–values irrespective of the genotype. Interestingly, after drilling the DHI related to untreated control discs of the respective disc levels and genotypes was significantly lower in C6-/- compared to C6+/- animals. This difference indicates that in the endplate drilling model the inability of TCC-formation may have some negative influence concerning preservation of disc height. Overall, endplate drilling could be successfully established as a new disc degeneration model in rabbits. Complete and persistent inability to generate a functional TCC, however, was not associated with an improved outcome after this type of injury.
Publications
- A possible role of the terminal complement complex TCC in intervertebral disc degeneration, ISSLS 2017, Athens, Greece
Neidlinger-Wilke C, Joos H, Saggese T, Brisby H, Barreto Henriksson H, Ruf M, Barth T, Mauer UM, Ignatius A, Brenner R
- Role of the terminal complement complex TCC in intervertebral disc degeneration, Philadelphia Spine Meeting 2017, Lake Harmony, Philadelphia, USA
Neidlinger-Wilke C, Joos H, Saggese T, Brisby H, Barreto Henriksson H, Ruf M, Barth T, Mauer UM, Ignatius A, Brenner R
- Regional differences in terminal complement complex (TCC) deposition suggest a correlation with degenerative changes of intervertebral disc endplates, International Conference on Complement Therapeutics 2018, Chania, Greece
Teixeira GQ, Saggese T, Yong Z, Kuhn A, Goncalves RM, Ruf M, Mauer UM, Ignatius A, Brenner R, Neidlinger- Wilke C
- (2019) Reduced terminal complement complex formation in mice manifests in low bone mass and impaired fracture healing. Am J Pathol. 189(1):147-161
Mödinger Y, Rapp AE, Vikman A, Ren Z, Fischer V, Bergdolt S, Haffner-Luntzer M, Song WC, Lambris JD, Huber-Lang M, Neidlinger- Wilke C, Brenner RE, Ignatius A
(See online at https://doi.org/10.1016/j.ajpath.2018.09.011) - Is the terminal complement complex a possible target for therapeutic innervation in intervertebral disc degeneration? Annual Meeting of the International Society for the Study of the Lumbar Spine (ISSLS) 2019, Kyoto, Japan
Teixeira GQ, Yong Z, Goncalves RM, Kuhn A, Ruf M, Nerlich A, Mauer UM, Ignatius A, Brenner R, Neidlinger-Wilke C
- Terminal complement complex (TCC): a possible target for in intervertebral disc degeneration therapeutics. Eurospine 2019, Helsinki, Finland
Teixeira GQ, Yong Z, Goncalves RM, Kuhn A, Ruf M, Nerlich A, Barth T, Mauer UM, Ignatius A, Brenner R, Neidlinger-Wilke C
(See online at https://doi.org/10.1007/s00586-019-06101-2) - Terminal complement complex formation in degenerated discs correlates with degree of degeneration. DWG 2019, Munich
Teixeira GQ, Yong Z, Goncalves RM, Kuhn A, Ruf M, Nerlich A, Mauer UM, Barth T, Ignatius A, Brenner R, Neidlinger-Wilke C
(See online at https://doi.org/10.1007/s00586-020-06630-1) - (2020) Crucial role of the terminal complement complex in chondrocyte death and hypertrophy after cartilage trauma. Osteoarthritis Cartilage. 28(5):685-697
Riegger J, Huber-Lang M, Brenner RE
(See online at https://doi.org/10.1016/j.joca.2020.01.004) - Can disc degeneration-associated factors activate terminal complement complex (TCC) formation? German Spine Conference (DWG) 2020, Virtual
Teixeira GQ, Yong Z, Kuhn A, Riegger J, Ruf M, Ignatius A, Brenner R, Neidlinger-Wilke C
(See online at https://doi.org/10.1007/s00586-020-06630-1) - IL-1β and Cathepsin-D modulate cellular terminal complement complex formation in human disc tissue. Orthopaedic Research Society (ORS) Annual Meeting 2020, Phoenix, Arizona, USA
Yong Z, Teixeira GQ, Kuhn A, Ruf M, Mauer UM, Ignatius A, Brenner R, Neidlinger-Wilke C
- Modulation of the terminal complement complex formation in human disc tissue cultures. Eurospine 2020, Virtual
Teixeira GQ, Yong Z, Kuhn A, Ruf M, Mauer UM, Ignatius A, Brenner R, Neidlinger-Wilke C
(See online at https://doi.org/10.1007/s00586-020-06683-2) - (2021) Interleukin-1β and cathepsin D modulate formation of the terminal complement complex in cultured human disc tissue. Eur Spine J. 30:2247-2256
Teixeira GQ, Yong Z, Kuhn A, Riegger J, Goncalves RM, Ruf M, Mauer UM, Huber-Lang M, Ignatius A, Brenner RE, Neidlinger-Wilke C
(See online at https://doi.org/10.1007/s00586-021-06901-5) - (2021) Mesenchymal stem cell secretome decreases the inflammatory response in annulus fibrosus organ cultures, Eur Cell Mater. 41:1-19
Neidlinger-Wilke C, Ekkerlein A, Goncalves RM, Ferreira JR, Ignatius A, Wilke HJ, Teixeira GQ
(See online at https://doi.org/10.22203/ecm.v042a01) - (2021) Terminal complement complex formation is associated with intervertebral disc degeneration. Eur Spine J. 30:217-226
Teixeira GQ, Yong Z, Goncalves RM, Kuhn A, Riegger-J, Brisby H, Henriksson HB, Ruf M, Barth T, Mauer UM, Ignatius A, Brenner R, Neidlinger-Wilke C
(See online at https://doi.org/10.1007/s00586-020-06592-4) - Deposition of terminal complement complex on intervertebral disc cells is associated with upregulation of MMP-1 and ADAMTS-4. Joint Meeting of the French and German Society of Matrix Biology 2021, Virtual
Kuhn A, Riegger J, Teixeira GQ, Ruf M, Neidlinger-Wilke C, Brenner R
- Inhibition of terminal complement complex deposition on intervertebral disc cells using eculizumab. DKOU 2021, Berlin, Germany
Kuhn A, Riegger J, Teixeira GQ, Ruf M, Neidlinger-Wilke C, Brenner R
(See online at https://doi.org/10.3205/21dkou393) - Interleukin-1β and cathepsin-D modulate the terminal complement complex formation in human disc tissue cultures. TERMIS World Congress 2021, Virtual
Teixeira GQ, Yong Z, Kuhn A, Riegger J, Ruf M, Ignatius A, Brenner R, Neidlinger-Wilke C
