O renascer da coluna: uma receita de inspiração fetal feita à medida para regenerar o disco intervertebral

IVD degeneration, is the leading cause of severe LBP in over 70% of the world population ranking first in terms of disability. Age-related cell decline and ECM changes trigger this debilitating condition for which current treatments lack efficacy. In addition, required treatment is often expensive, lengthy, and very invasive rarely treating the underlying disease, which frequently translates in recurrent episodes of pain and reoperations. Indeed, one of the major CHALLENGES in the field is to be able to restore a functional IVD, in order to promote a long-term regenerative effect in a minimally invasive way. To overcome the drawbacks of existing strategies REBORN_DISC will develop a revolutionary strategy to treat IVD degeneration. Project MAIN OBJECTIVES are: 1) to pursue the development of a superior fetal cell-derived matrix (fMSC-DM), using CRISPR engineering to target COL12A1 and COL14A1 activation in Mesenchymal Stem Cells (MSCs) and 2) unravel the mechanisms behind fetal matrix-cell communication, responsible for maintaining tissue homeostasis. THE HYPOTHESIS behind this ambitious proposal is that the fMSC-DM that recapitulates an early stage microenvironment will rejuvenate human degenerated NP cells, promoting endogenous repair by favouring the maintenance, proliferation and differentiation of adult NP stem/progenitor cell niches. To reach the above goals, the PLAN encompasses: a) generation of simple and efficient biofactories of fMSC-DM production by CRISPR engineering of MSCs; b) characterization of the biochemical, biomechanical and ultrastructural profile of the fMSC-DM; c) evaluation of the capacity of this engineered matrix to maintain human NP progenitors and to rejuvenate patient derived NP cells; d) in vivo validation of the potential of the fMSC-DM to regenerate the IVD and solve LBP in rats. The greatest NOVELTY of this project is the cell-derived matrix fetal tailoring by CRISPR engineering. Cell-derived matrices are an attractive alternative to surpass limitations of animal-derived scaffolds, circumventing zoonosis, reducing heterogeneity and overcoming donor shortage. In addition by being genetically customized to mimic a prenatal microenvironment through fetal gene reactivation, this fetal- inspired ECM will boost tissue regeneration favoring proliferation, differentiation and matrix synthesis of adult cells and resident progenitors, potentiating endogenous repair. This solution based on cutting-edge technology, also holds potential to improve the already promising MSC-based therapies. The huge gains on fine tuning this genetically tailored fetal matrix will go significantly BEYOND CURRENT KNOWLEDGE, having a broader impact on other fields, namely those of tissue regeneration in general and oncology. The immense benefit of such cell-based/cell-free solutions stems from removing the risks faced when using genetically modified cells themselves. All in all, REBORN_DISC will pave the way for human intervention and for a successful LBP treatment, thus promoting healthy and active ageing opening new avenues for other far-reaching applications in cartilage, skin, cornea and heart regeneration among others.