Revelação de alvos imunomoduladores secretos no disco intervertebral para tratar a dor lombar

The IMMUNOSECRET project has the ambition of revealing novel therapeutic targets for LBP and revolutionize the field of anti-inflammatory drugs for this disorder, leader in years lived with disability and rehabilitation services and an enormous burden to healthcare systems worldwide. This is an enormous challenge, that is fully aligned with the goal 3 of ONU Agenda 2030 for Sustainable Development: “Ensure healthy lives and promote well-being for all at all ages”. Our team has been gathering knowledge on the relevance of inflammation in IVD degeneration. In particular, the crucial role of IL-1b, as the most prevalent pro-inflammatory mediator in LBP (Risbud & Shapiro, 2014). IMMUNOSECRET hypothesizes that inflammation activates myeloid-derived IVD cells, crucial for disease progression, and that tackling specifically these cells with immunomodulatory therapies, we can stop/revert IVD degenerative cascade. This hypothesis goes beyond the state of the art in IVD biology. To test the hypothesis, a combination of knowledge in immunology, cell biology, orthopedics, biomaterials and bioengineering will be aligned to address the following objectives: 1) To disclose a signature of IVD myeloid-derived cells Resident IVD tissue-myeloid cells will be identified in the bovine (healthy) IVD, using a multi-color flow cytometry panel of surface markers, previously optimized for bovine adipose tissue (Oliveira et al., 2020). Bovine will overcome lack of human healthy IVD and immune cell infiltration in degenerated IVD. IVD myeloid-derived cells spatial transcriptomics and proteomics profile will be traced compared with the non-myeloid cell fraction. 2) To engineer a 3D microenvironment to maintain/expand IVD myeloid-derived cells in vitro Hydrogels based on decellularized extracellular matrix (dECM) from healthy bovine IVD, as recently been established (Fiordalisi et al., 2022), with adequate structural, biochemical and biomechanical properties, will be developed by a team with strong expertise in hydrogels that recapitulate native ECM (Neves et al., 2020), to create robust models to study IVD cell subpopulations, as IVD myeloid-derived cells. 3) To understand the function of IVD myeloid-derived cells in disease progression The response of IVD myeloid-derived cells to degenerative microenvironment, i.e. pro-inflammatory environment and mechanical loaded, that act in synergy, promoting IVD catabolism (Saggese T, 2019), will be investigated by a team expert in inflammation and bioreactors (Gonçalves, 2022). Cell response in terms of production of ECM, pro-inflammatory, pro-angiogenic and neurotrophic factors will allow to understand the role of these cells in disease progression. 4) To screen anti-inflammatory/immunomodulatory therapies on IVD myeloid-derived cells to stop/revert disease progression The effects of non-steroidal anti-inflammatory (NSAID) drugs, MSC-based immunomodulatory products (secretome, EVs) or alternative drugs will be screened in IVD-myeloid derived cells, as used by the team in other contexts (Cunha et al., 2017; Ferreira et al., 2021; Pereira et al., 2016; Teixeira, Leite Pereira, et al., 2016; Teixeira et al., 2018). Primed MSC secretome/EVs has demonstrated a superior immunomodulatory capacity and will be tested here, expecting to validate the relevance of IVD myeloid-derived cells in IVD degeneration.. Overall, this project will uncover novel targets for LBP, bringing new light to immunomodulatory therapies for LBP.