Reprogramação de astrócitos corticais em neurônios utilizando coquetel de pequenas moléculas

Abstract

Recently, many techniques have been described to generate neurons that could be used in cell-based therapies to neurodegenerative diseases or acute central nervous system injuries. One of such techniques is the lineage reprogramming of differentiated cells into neurons, either directly or indirectly. However, most protocols for lineage reprogramming rely on the expression of ectopic genes, which could lead to other unexpected transcriptional changes in the reprogrammed cells. To circumvent this possibility, we here evaluate the possibility to reprogram astrocytes from post-natal mouse cortex into neurons through the use of a cocktail of small molecules in the cell culture medium. This cocktail contains molecules associated with different cellular pathways, including gene expression regulation, neurogenesis modulation and cell cycle control, and it has been previously shown to reprogram fibroblasts into neurons. To assess the cellular phenotype after treatment of astrocytes cultures with small molecules, we evaluated the expression of neuron-specific proteins, morphology and gene expression by immunocytochemistry and RT-qPCR. Our preliminary data show that astrocytes adopt morphologies and gene expression compatible with a neuronal phenotype, indicating that small molecules can be used to reprogram astroytes into induced neurons.