POLYMERIC STABILIZED REVERSE MICELLES FOR GALC BRAIN DELIVERY IN KRABBE DISEASE TREATMENT

Krabbe disease (KD) is a severe disorder leading to oligodendrocytes loss, demyelination, and neuroinflammation with detrimental effects on health for those affected. KD is encompassed within the category of lysosomal storage disorders (LSDs), due to enzymatic defects/deficiencies within the lysosomes. In particular, KD is caused by mutations in the galactosylceramidase (GALC) enzyme, provoking the accumulation of the neurotoxic metabolite psychosine^​. The enzyme replacement therapy (ERT), consisting in the systemic administration of the recombinant functional enzyme, is one of the approaches that can be beneficial for some LSDs not primarily affecting the brain. Although not able to cure cerebral damage, ERT can weaken the symptoms and decelerate the worsening of the pathology^​. Previously, we successfully developed stabilized reversed polymeric micelles (SRMs) as carrier of hydrophilic macromolecules^​. Here, we test this nanotechnology as potential vector for GALC delivery in KD. GALC-loaded SRMs were produced and characterized by size, surface charge and enzymatic activity, showing physico-chemical characteristics suitable for brain delivery through systemic administration. We performed preliminary in vitro experiments on oligodendrocytes to ascertain SRMs cytotoxicity by WST-1 assay, and to characterize cell internalization via advanced microscopy analysis. Our results indicate that SRMs are non-toxic, and that they are conveyed into the lysosomes 24h post-administration. In addition, we evaluated the recovery of enzymatic activity in GALC-knock-out oligodendrocytes as in vitro model of KD, revealing enzymatic activity retrieval. In conclusion, GALC-loaded SRMs are biocompatible and ensure enzyme stability, representing a promising tool for ERT in KD. This investigation is part of REMEDY project (FISA2022-00627).