title: Potential Transfer of Polyglutamine and CAG-Repeat RNA in Extracellular Vesicles in Huntington's Disease: Background and Evaluation in Cell Culture.
abstract: In Huntington's disease (HD) the imperfect expanded CAG repeat in the first exon of the HTT gene leads to the generation of a polyglutamine (polyQ) protein, which has some neuronal toxicity, potentially mollified by formation of aggregates. Accumulated research, reviewed here, implicates both the polyQ protein and the expanded repeat RNA in causing toxicity leading to neurodegeneration in HD. Different theories have emerged as to how the neurodegeneration spreads throughout the brain, with one possibility being the transport of toxic protein and RNA in extracellular vesicles (EVs). Most cell types in the brain release EVs and these have been shown to contain neurodegenerative proteins in the case of prion protein and amyloid-beta peptide. In this study, we used a model culture system with an overexpression of HTT-exon 1 polyQ-GFP constructs in human 293T cells and found that the EVs did incorporate both the polyQ-GFP protein and expanded repeat RNA. Striatal mouse neural cells were able to take up these EVs with a consequent increase in the green fluorescent protein (GFP) and polyQ-GFP RNAs, but with no evidence of uptake of polyQ-GFP protein or any apparent toxicity, at least over a relatively short period of exposure. A differentiated striatal cell line expressing endogenous levels of Hdh mRNA containing the expanded repeat incorporated more of this mRNA into EVs as compared to similar cells expressing this mRNA with a normal repeat length. These findings support the potential of EVs to deliver toxic expanded trinucleotide repeat RNAs from one cell to another, but further work will be needed to evaluate potential EV and cell-type specificity of transfer and effects of long-term exposure. It seems likely that expanded HD-associated repeat RNA may appear in biofluids and may have use as biomarkers of disease state and response to therapy.
authors.affiliation: Molecular Neurogenetics Unit, Department of Neurology, Massachusetts General Hospital-East, 13th Street, Building 149, Charlestown, MA, 02129, USA.|Center for Molecular Imaging Research, Department of Radiology, Massachusetts General Hospital, Boston, MA, USA.|Center for NeuroDiscovery, Harvard Medical School, Boston, MA, USA.
authors.forename: Xuan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Molecular Neurogenetics Unit, Department of Neurology, Massachusetts General Hospital-East, 13th Street, Building 149, Charlestown, MA, 02129, USA.|Center for Molecular Imaging Research, Department of Radiology, Massachusetts General Hospital, Boston, MA, USA.|Center for NeuroDiscovery, Harvard Medical School, Boston, MA, USA.
authors.forename: Erik R
authors.initials: ER
authors.lastname: Abels
authors.affiliation: Department of Pharmaceutical Sciences, University of Colorado Denver Skaggs School of Pharmacy and Pharmaceutical Sciences, Aurora, CO, USA.
authors.forename: Jasmina S
authors.initials: JS
authors.lastname: Redzic
authors.affiliation: Gladstone Institute of Neurological Disease and the Departments of Neurology and Physiology, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Julia
authors.initials: J
authors.lastname: Margulis
authors.affiliation: Gladstone Institute of Neurological Disease and the Departments of Neurology and Physiology, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Steve
authors.initials: S
authors.lastname: Finkbeiner
authors.affiliation: Molecular Neurogenetics Unit, Department of Neurology, Massachusetts General Hospital-East, 13th Street, Building 149, Charlestown, MA, 02129, USA. breakefield@hms.harvard.edu.|Center for Molecular Imaging Research, Department of Radiology, Massachusetts General Hospital, Boston, MA, USA. breakefield@hms.harvard.edu.|Center for NeuroDiscovery, Harvard Medical School, Boston, MA, USA. breakefield@hms.harvard.edu.
authors.forename: Xandra O
authors.initials: XO
authors.lastname: Breakefield
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D000067128:Extracellular Vesicles / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D000071058:Huntingtin Protein / Q000378:metabolism; D006816:Huntington Disease / Q000378:metabolism*; D051379:Mice; D017072:Neostriatum / Q000378:metabolism / Q000473:pathology; D010455:Peptides / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D019680:Trinucleotide Repeat Expansion / Q000235:genetics*
chemical_list: D000071058:Huntingtin Protein; D010455:Peptides; D049452:Green Fluorescent Proteins; C097188:polyglutamine; D012313:RNA
keywords: Exosomes; Huntington’s disease; Neurodegeneration; Trinucleotide repeat