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A single-cell map of antisense oligonucleotide activity in the brain
Meredith A Mortberg1, Juliana E Gentile1, Naeem M Nadaf1
1Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
Nucleic Acids Research
|May 15, 2023
Summary
Antisense oligonucleotides (ASOs) delivered to the central nervous system (CNS) show broad cell type activity. Single nucleus sequencing reveals ASO effects in all cells, linking CSF biomarkers to neuronal activity for CNS disorder treatments.
Area of Science:
- Neuroscience
- Molecular Biology
- Pharmacology
Background:
- Antisense oligonucleotides (ASOs) offer potential for treating central nervous system (CNS) disorders by modulating RNA.
- Understanding ASO biodistribution and activity across diverse CNS cell types is crucial for therapeutic development.
- Current assessments often rely on bulk tissue analysis and limited biomarker readouts.
Purpose of the Study:
- To investigate the cell type-specific distribution and activity of centrally delivered ASOs in the CNS.
- To correlate ASO activity in individual cells with bulk tissue measurements and cerebrospinal fluid (CSF) biomarkers.
- To establish single nucleus sequencing as a method for evaluating oligonucleotide therapeutic efficacy.
Main Methods:
- Single nucleus RNA sequencing was performed on CNS tissue from mice and non-human primates (NHPs) treated with ASOs targeting Prnp and Malat1.
- Pharmacologic activity and target RNA suppression were quantified across various CNS cell types.
- CSF biomarker levels (PrP) were measured in NHPs treated with a PRNP-targeting ASO.
Main Results:
- ASO pharmacologic activity was detected in all analyzed CNS cell types, with varying magnitudes.
- Target RNA suppression was observed in every sequenced cell, indicating widespread cellular engagement.
- Duration of ASO action differed across cell types, with shorter effects in microglia compared to neurons.
- Neuronal ASO suppression was comparable to or stronger than bulk tissue levels.
- In NHPs, CSF PrP reduction correlated with PRNP knockdown across all cell types, including neurons.
Conclusions:
- Centrally delivered ASOs engage a broad spectrum of CNS cell types, including neurons and microglia.
- Single nucleus sequencing provides a high-resolution method to assess cell type-specific ASO activity.
- CSF biomarkers can reflect ASO pharmacodynamic effects in disease-relevant cells, supporting their use in clinical trials for neuronal disorders.
