Neurotropism of SARS-CoV-2 towards differentiated neuronal SH-SY5Y cells and neural progenitor cell line ReNcell

  • Matthieu GUEIBE

Student thesis: Master typesMaster en sciences biomédicales à finalité spécialisée en recherche préclinique

Résumé

The world has been recently facing a pandemic caused by severe acute syndrome coronavirus 2 (SARS-CoV-2), responsible for the Coronavirus disease 2019 (COVID-19). Although it manifests as flu-like symptoms, there is increasing evidence that neurological-related symptoms (anosmia, ageusia, cognitive impairment) emerge during infection but also persist even after the disease has resolved (long-COVID phase). SARS-CoV-2 could therefore reach the central nervous system and disturb some of its functions. To model such neurotropism, one of the aims of this work was to characterize the cytopathic effects caused by SARS-CoV-2 on a neuron-like cell line, SH-SY5Y differentiated into dopaminergic-like cells and whose susceptibility has been previously demonstrated. Infection of these cells caused a significant reduction in the length of their neuritic processes, a decrease in the release of neuropeptide Y and occurrence of plasma membrane eruptions. No difference in the expression of cytoskeletal proteins such as β-III tubulin and tau protein, or even in their post-translational modifications was evidenced using western blot technique. No mis-location of β-III tubulin and tau protein or abnormal expression have been observed in double immunofluorescence experiments. Finally, infection did not compromise the cell viability. In order to counteract the cytopathic effects, five entry inhibitors, targeting TMPRSS2 (camostat mesylate), spike protein (anti-RBD antibody and human recombinant soluble ACE2), CD147 (anti-CD147 antibody) and NRP1 receptors (EG00229) were co-incubated with the viral inoculum. RT-qPCRs on the viral E and RdRp genes showed partial inhibition of entry with anti-RBD antibody and EG00229. Secondly, neuritic processes atrophy could be reversed in the presence of each inhibitor at different concentrations. Finally, higher concentrations of camostat mesylate and hrs ACE2 restored neuropeptide Y secretion to basal levels. In the second aim of this work, the neurotropism of SARS-CoV-2 towards a neural progenitor cell line, ReNcells, was studied, the effect on its differentiation abilities. The ReNcells, grown as undifferentiated cells, were prone to SARS-CoV-2 infection at high MOI, although not reaching statistical significance. Viral replication appeared to occur in a limited number of cells. Based on our preliminary data on protein expression and cell morphology, it was not possible to demonstrate or refute that SARS-CoV-2 had any impact on the differentiation of these cells. In conclusion, SARS-CoV-2 was able to induce mild but significant cytopathic effects on differentiated SH-SY5Y cells, which can be partly prevented by entry inhibitors. Additionally, other investigations are needed to determine whether SARS-CoV-2 has an impact on neural stem cell differentiation.
la date de réponse17 janv. 2023
langue originaleAnglais
L'institution diplômante
  • Universite de Namur
SuperviseurCharles Nicaise (Promoteur) & Nicolas Gillet (Copromoteur)

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