An RNA-targeting CRISPR-Cas13d system alleviates disease-related phenotypes in Huntington's disease models

Kathryn H Morelli; Qian Wu; Maya L Gosztyla; Hongshuai Liu; Minmin Yao; Chuangchuang Zhang; Jiaxu Chen; Ryan J Marina; Kari Lee; Krysten L Jones; Megan Y Huang; Allison Li; Charlene Smith-Geater; Leslie M Thompson; Wenzhen Duan; Gene W Yeo

doi:10.1038/s41593-022-01207-1

An RNA-targeting CRISPR-Cas13d system alleviates disease-related phenotypes in Huntington's disease models

Nat Neurosci. 2023 Jan;26(1):27-38. doi: 10.1038/s41593-022-01207-1. Epub 2022 Dec 12.

Authors

Kathryn H Morelli^#¹, Qian Wu^#^{2

3}, Maya L Gosztyla¹, Hongshuai Liu², Minmin Yao², Chuangchuang Zhang², Jiaxu Chen³, Ryan J Marina¹, Kari Lee¹, Krysten L Jones¹, Megan Y Huang¹, Allison Li¹, Charlene Smith-Geater⁴, Leslie M Thompson^{4

5}, Wenzhen Duan^{6

7

8}, Gene W Yeo^{9

10

11}

Affiliations

¹ Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA.
² Division of Neurobiology, Department of Psychiatry and Behavioral Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
³ School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China.
⁴ Department of Psychiatry and Human Behavior, University of California Irvine, Irvine, CA, USA.
⁵ Department of Neurobiology and Behavior, University of California Irvine, Irvine, CA, USA.
⁶ Division of Neurobiology, Department of Psychiatry and Behavioral Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, USA. [email protected].
⁷ The Solomon H Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA. [email protected].
⁸ Program in Cellular and Molecular Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA. [email protected].
⁹ Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA. [email protected].
¹⁰ Institute for Genomic Medicine, University of California San Diego, La Jolla, CA, USA. [email protected].
¹¹ Stem Cell Program, University of California San Diego, La Jolla, CA, USA. [email protected].

^# Contributed equally.

Abstract

Huntington's disease (HD) is a fatal, dominantly inherited neurodegenerative disorder caused by CAG trinucleotide expansion in exon 1 of the huntingtin (HTT) gene. Since the reduction of pathogenic mutant HTT messenger RNA is therapeutic, we developed a mutant allele-sensitive CAG^EX RNA-targeting CRISPR-Cas13d system (Cas13d-CAG^EX) that eliminates toxic CAG^EX RNA in fibroblasts derived from patients with HD and induced pluripotent stem cell-derived neurons. We show that intrastriatal delivery of Cas13d-CAG^EX via an adeno-associated viral vector selectively reduces mutant HTT mRNA and protein levels in the striatum of heterozygous zQ175 mice, a model of HD. This also led to improved motor coordination, attenuated striatal atrophy and reduction of mutant HTT protein aggregates. These phenotypic improvements lasted for at least eight months without adverse effects and with minimal off-target transcriptomic effects. Taken together, we demonstrate proof of principle of an RNA-targeting CRISPR-Cas13d system as a therapeutic approach for HD, a strategy with implications for the treatment of other dominantly inherited disorders.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Clustered Regularly Interspaced Short Palindromic Repeats
Corpus Striatum / metabolism
Disease Models, Animal
Huntingtin Protein / genetics
Huntingtin Protein / metabolism
Huntington Disease* / genetics
Huntington Disease* / metabolism
Huntington Disease* / therapy
Mice
Phenotype
RNA
RNA, Messenger / metabolism

Substances

RNA
RNA, Messenger
Huntingtin Protein

Abstract

Publication types

MeSH terms

Substances

Grants and funding