Avaliação do tratamento com VU0409551 em astrócitos derivados de hiPSCs estimulados com citocinas pró-inflamatórias associadas à Esclerose Múltipla

Abstract

Multiple sclerosis (MS) is a neuroinflammatory, autoimmune, chronic and disabling disease that affects the central nervous system due to an autoimmune response against myelin proteins, along with the formation of inflammatory infiltrate, demyelination, excitotoxicity, astrogliosis and neurodegeneration. Astrocytes are the most abundant cell type in the mammalian central nervous system (CNS) and, upon injury, they become reactive. Activation of metabotropic glutamate receptor subtype 5 (mGluR5) has been shown to be neuroprotective upon glutamate insult and can inhibit pro-inflammatory microglial signaling in vitro, as well as induce microglial and astrocytic BDNF expression in a murine model of MS. Currently, there are no therapeutic approache that could mitigate both MS-related neuroinflamation and neurogeneration. In the present study, we aimed to establish a pro-inflammatory stimulus with cytokines associated with MS pathology and astrocytic reactivity, tumor necrosis factor alpha (TNF -α), interleukin 1 beta (IL-1β) and granulocyte and macrophage colony stimulating factor (GM-CSF) and to evaluate the effect of the positive modulator (PAM) of mGluR551, VU0409551, in stimulated astrocytes derived from human induced pluripotency cells (hiPSCs). We observed an increase in the expression of TNF-α and IL-1β 24 hours following TNF-α and IL-1β stimulation. VU0409551 was found to increase the expression of IL-6, regardless of TNF-α and IL-1β stimulation. Furthermore, a marginal effect of treatment with 1μM of VU0409551 was observed in the prevention of the elevation of IL-1β expression induced by the pro-inflammatory stimulus with TNF-α and IL-1β over 24h. The results indicate that TNF-α, IL-1β and VU0409551 treatment does not change the expression of the other evaluated targets IL-10, GNR, BDNF, EAAT1, and does not modify glutamate levels in astrocyte conditioned media. Therefore, the assumed anti- inflammatory effect of VU0409551 was not confirmed by the results in this study.