Examining the role of glycosphingolipids in the progression and heterogeneity of synucleinopathies

Project: Research project

Description

Parkinson's disease (PD) is characterized by the presence of Lewy body inclusions in the nervous system comprised of insoluble alpha-synuclein (a-syn). The mechanisms that dictate the conversion of a-syn from its physiological state into pathogenic aggregates are not completely understood. Genetic studies have provided clues into the etiology of PD and causative factors that might promote a-syn inclusions. Among these, mutations in the lysosomal system have emerged as key risk factors for PD. Since lysosomes can degrade physiological a-syn, loss-of-lysosomal function may promote a-syn accumulation and drive aggregation. Mutations in lysosomal GBA1 have been identified as a strong risk factor for PD, and are highly associated with early onset dementia in PD patients. GBA1 mutations are also strong risk factors for a related synucleinopathy, Dementia with Lewy bodies (odds ratio=8.28), that often involves the co-aggregation of other disease-linked proteins linked to Alzheimer's disease such as tau and A-beta. GBA1 encodes lysosomal beta-Glucocerebrosidase (GCase), which degrades glycosphingolipids (GSLs) such as glucosylceramide (GluCer) in lysosomes. Homozygous GBA1 mutations cause the rare lysosomal storage disorder, Gaucher's disease (GD), which is also characterized by Lewy body inclusions in the nervous system. All of the GD or PD associated GBA1 mutations identified to date have been well characterized and cause loss-of-enzymatic function. Additionally, GluCer substrate accumulation has been documented in patient-derived tissues and iPSC-derived neurons of both diseases. We previously showed that GluCer directly converts physiological a-syn conformers into pathogenic assembly-state species in GD iPSC-neuronal models and patient derived midbrain neurons. Here, we propose to study the mechanism of GluCer-induced aggregation by examining the how aggregates propagate and persist in cell free, cell culture, and mouse models. Pathogenic a-syn oligomers persist for hundreds of days in patient-derived iPSC-neurons, and our studies will examine potential mechanisms to explain this phenomenon. We will determine if the unique conformational information of GluCer-induced oligomers can be replicated by interaction and conversion of physiological a-syn conformers, their ability to cross-seed aggregates with tau or A-beta, neuron-to-neuron dissemination, and transmission of metabolic dysfunction. It is well known that PD and other synucleinopathies demonstrate remarkable heterogeneity for which there is no mechanistic explanation. Our studies will begin to examine this process by testing the cross-seeding potential of GluCer-induced a-syn oligomers. We plan to study these processes in human iPSC-midbrain models, which should provide a highly relevant disease model since they are derived directly from PD or GD patients. To supplement the iPSC studies, we will employ primary neuronal cultures, established cell lines, and mouse models of GD to test our hypotheses in vivo. We will relate GluCer-induced changes in pathological a-syn to comorbid pathologies and neuronal toxicity in vitro and in vivo. If successful, these studies will provide novel insight into the mechanisms of how loss of GBA1 function leads to PD and dementia. Our studies may contribute more broadly to protein aggregation diseases, by determining if the unique conformational features of a-syn, identified in our lab, can be transferred to newly made, physiological a-syn. We will determine if these unique structures can cross-seed A-beta and tau, providing a novel explanation for how
StatusActive
Effective start/end date3/1/192/29/24

Funding

  • National Institute of Neurological Disorders and Stroke (1R01NS109157-01)

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Glycosphingolipids
Glucosylceramides
Parkinson Disease
Gaucher Disease
Neurons
Mutation
Lewy Bodies
Mesencephalon
Lysosomes
Nervous System
Dementia
Seeds
Glucosylceramidase
Lewy Body Disease
alpha-Synuclein
Aptitude
Alzheimer Disease
Proteins
Cell Culture Techniques
Odds Ratio