A novel glucosylceramide synthase inhibitor attenuates alpha synuclein pathology and lysosomal dysfunction in preclinical models of synucleinopathy.

Abstract

Mutations in the lysosomal enzyme glucocerebrosidase (GCase, GBA1 gene) are the most common genetic risk factor for developing Parkinson's disease (PD). GCase metabolizes the glycosphingolipids glucosylceramide (GlcCer) and glucosylsphingosine (GlcSph). Mutations in GBA1 reduce enzyme activity and the resulting accumulation of glycosphingolipids may contribute to the underlying pathology of PD, possibly via altering lysosomal function. While reduction of GCase activity exacerbates α-synuclein (α-syn) aggregation, it has not been determined that this effect is the result of altered glycosphingolipid levels and lysosome function or some other effect of altering GCase. The glycosphingolipid GlcCer is synthesized by a single enzyme, glucosylceramide synthase (GCS), and small molecule inhibitors (GCSi) reduce cellular glycosphingolipid levels. In the present studies, we utilize a preformed fibril (PFF) rodent primary neuron in vitro model of α-syn pathology to investigate the relationship between glycosphingolipid levels, α-syn pathology, and lysosomal function. In primary cultures, pharmacological inhibition of GCase and D409V GBA1 mutation enhanced accumulation of glycosphingolipids and insoluble phosphorylated α-syn. Administration of a novel small molecule GCSi, benzoxazole 1 (BZ1), significantly decreased glycosphingolipid concentrations in rodent primary neurons and reduced α-syn pathology. BZ1 rescued lysosomal deficits associated with the D409V GBA1 mutation and α-syn PFF administration, and attenuated α-syn induced neurodegeneration of dopamine neurons. In vivo studies revealed BZ1 had pharmacological activity and reduced glycosphingolipids in the mouse brain to a similar extent observed in neuronal cultures. These data support the hypothesis that reduction of glycosphingolipids through GCS inhibition may impact progression of synucleinopathy and BZ1 is useful tool to further examine this important biology.

• Authors: Mali Cosden, Sarah Jinn, Lihang Yao, Cheryl A Gretzula, Monika Kandebo, Dawn Toolan, Nathan G Hatcher, Lei Ma, Wei Lemaire, Gregory C Adam, Christine Burlein, Christina Minnick, Rose Flick, Marla L Watt, James Mulhearn, Mark Fraley, Robert E Drolet, Jacob N Marcus, Sean M Smith
• Journal: Neurobiology of disease (Neurobiol Dis) Vol. 159 Pg. 105507 (11 2021) ISSN: 1095-953X [Electronic] United States
• PMID: 34509608 (Publication Type: Journal Article)
• Copyright: Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.

Chemical References

• Benzoxazoles
• Glycosphingolipids
• Protein Aggregates
• alpha-Synuclein
• Glucosyltransferases
• ceramide glucosyltransferase
• Glucosylceramidase

Topics

• Animals
• Benzoxazoles (pharmacology)
• Dopaminergic Neurons (drug effects, metabolism)
• Glucosylceramidase (genetics)
• Glucosyltransferases (antagonists & inhibitors)
• Glycosphingolipids (metabolism)
• In Vitro Techniques
• Lysosomes (drug effects, metabolism)
• Mice
• Neurons (drug effects, metabolism)
• Parkinson Disease (genetics, metabolism)
• Primary Cell Culture
• Protein Aggregates
• Rats
• Synucleinopathies (genetics, metabolism)
• alpha-Synuclein (drug effects, metabolism)