D-003 | In vitro and in vivo effects of the intransasal administration of extracellular vesicles loaded with the neuronal glycoprotein M6a on chronic stress related disorders.

Cellular and Molecular Neurobiology
Author: María Victoria Bühler | Email: mvbuhler@estudiantes.unsam.edu.ar

María Victoria Bühler^1°2°3°, Camila Attonaty^1°2°3°, Marcela Brocco^1°2°3°,  Melisa Carolina Monteleone^1°2°3°

^1° Instituto de Investigaciones Biotecnológicas IIBIO
^2° Escuela de Bio y Nanotecnologías EByN
^3° Universidad Nacional de General San Martín UNSAM

Chronic stress is involved in the etiology of various pathologies, including depression. Due to the heterogeneity of symptoms, existing treatments present low remission rates and undesirable side effects. Therefore, an alternative strategy is proposed: the administration of extracellular vesicles (EVs) containing therapeutic factors added in vitro. Since M6a is a glycoprotein that contributes to neuronal plasticity and its levels are reduced under stress conditions, we propose using EVs loaded with the M6a plasmid for intranasal administration in mice exposed to chronic restraint stress. This approach will allow us to evaluate whether some of the behavioral and/or molecular phenotypes associated with the pathology are reversed.
First, we standardized the isolation of EVs from the HT22 cell line through ultracentrifugation and characterized them using Western blot, TEM, and NTA. Next, we loaded the EVs and analyzed potential changes in their surface. Finally, we administered the modified EVs to C57 mice subjected to a restraint stress protocol and evaluated their behavior through sucrose preference and forced swim tests, their weight and M6a expression in the hippocampus. Our results indicate that we obtained particle populations of approximately 125 nm in diameter with morphology and sizes compatible with EVs. Animals treated with loaded EVs, unlike those treated with PBS, showed differences in weight, in the forced swim test and in M6a expression.