A regenerative approach to the treatment of Multiple Sclerosis: identification of drug targets

Remyelination is the process that restores the myelin sheath after a degenerative process, or a viral or auto-immune attack. This process occurs throughout life in the Central Nervous System (CNS) of healthy individuals, thanks to the proliferation, migration and differentiation of oligodendrocyte precursor cells (OPCs). Despite the process underlying OPCs repopulation in the damaged area is actively studied, the molecular mechanisms are still unclear. In aged individuals or during the secondary phase of Relapsing Remitting Multiple Sclerosis (RRMS), the inefficiency of the process leads to permanent disability. Several immune-modulatory treatments that limit neuronal damage in RRMS patients are now available but none of these stimulate CNS remyelination efficiently. Novel pharmacological remyelination approaches are conceived as co-therapies. In order to reverse neuronal damage and improve patients’ standard of living, we screened a Prestwick chemical library for the property to upregulate myelin basic protein (MBP) in order to identify promyelinating drugs with known biological activity to combine with immunomodulatory therapy. The identification of pharmacological target(s) for small molecules screened and selected for a specific property, is essential. For this reason, the aim of this work is to validate the hypothetical targets previously identified by chemical-protein interactome of selected pro-myelinating drugs using OlineuM cellular model. A high-content screening of agonists and antagonists receptors potentially involved in the myelination, as well as the evaluation of perturbations at network level, allowed to reveal the molecular mechanisms underlying the cross-talk of protein complex networks, in the oligodendrocyte differentiation and myelination process. Additionally, we identified combined treatments that highly increased MBP expression and strongly induced morphological changes in cell shape. We took advantage of 3D polystyrene microfibers mimicking neurons for validating the efficacy of this drugs and observing the neuronal engagement.