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Published on: March 12, 2018
In vivo CRISPR base editing for treatment of Huntington's disease
Shraddha Shirguppe1, Michael Gapinske1,2, Devyani Swami1
1Department of Bioengineering, The Grainger College of Engineering, University of Illinois Urbana-Champaign, Urbana, IL, USA.
Insights
CRISPR base editing successfully reduced toxic Huntington
Area of Science:
- Neuroscience
- Genetics
- Molecular Biology
Background:
- Huntington's disease (HD) is a fatal neurodegenerative disorder.
- It is caused by an expanded CAG repeat in the huntingtin (HTT) gene.
- Proteolytic cleavage of mutant HTT produces toxic fragments, a key pathogenic event.
Purpose of the Study:
- To develop CRISPR base editors to create proteolysis-resistant HTT.
- To disrupt the splice acceptor of HTT exon 13, which contains cleavage sites.
Main Methods:
- CRISPR base editors were designed to target HTT exon 13.
- Editors were delivered to the striatum of a rodent model of HD.
- HTT fragment formation, aggregation, and behavioral deficits were assessed.
Main Results:
- Base editing reduced the formation of toxic HTT fragments.
- HTT aggregation and neuronal dysfunction were decreased.
- Functional deficits and brain atrophy were attenuated in the HD model.
Conclusions:
- CRISPR base editing targeting HTT exon 13 is a potential therapeutic strategy for HD.
- This approach mitigates mutant HTT toxicity by preventing fragment production.
- Splice-site modulation offers a promising avenue for treating Huntington's disease.
Abstract:
Huntington's disease (HD) is a fatal neurodegenerative disorder caused by an expanded CAG repeat within exon 1 of the huntingtin (HTT) gene, resulting in a mutant protein that drives neuronal dysfunction and loss. A key event in the pathogenesis of HD is proteolytic cleavage of mutant HTT, which generates aggregation-prone N-terminal fragments that contribute to toxicity. Strategies that prevent this process thus hold therapeutic potential. Here we develop CRISPR base editors that generate proteolysis-resistant HTT isoforms by disrupting the splice acceptor of HTT exon 13, an exon that encodes critical proteolytic cleavage sites implicated in N-terminal fragment production. When delivered to the striatum of an HD rodent model, these editors reduced HTT fragment formation, decreased aggregation, improved functional deficits and attenuated brain atrophy. Collectively, these results demonstrate the potential of base editing and splice-site modulation to mitigate mutant HTT toxicity in HD.
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