Allele-specific RNA interference prevents neuropathy in Charcot-Marie-Tooth disease type 2D mouse models.

Authors

Kathryn H Morelli, Jackson Laboratory & University of Maine
Laurie B Griffin, University of Michigan
Nettie K Pyne, Center for Gene Therapy, Research Institute at Nationwide Children's Hospital
Lindsay M Wallace, Center for Gene Therapy, Research Institute at Nationwide Children's Hospital
Allison M Fowler, Center for Gene Therapy, Research Institute at Nationwide Children's Hospital
Stephanie N Oprescu, The University Of Michigan
Ryuichi Takase, Thomas Jefferson University
Na Wei, Scripps Research Institute
Rebecca Meyer-Schuman, University of Michigan
Dattatreya Mellacheruvu, University of Michigan
Jacob O Kitzman, University of Michigan
Samuel G Kocen, Jackson Labratory
Timothy J Hines, Jackson Laboratory
Emily L Spaulding, Jackson Laboratory & University of Maine
James R Lupski, Baylor College of Medicine & Texas Children's Hospital
Alexey Nesvizhskii, University of Michigan
Pedro Mancias, University of Texas Health Science Center at Houston
Ian J Butler, University of Texas Health Science Center at Houston
Xiang-Lei Yang, Scripps Research Institute
Ya-Ming Hou, Thomas Jefferson UniversityFollow
Anthony Antonellis, University Michigan
Scott Q Harper, Center for Gene Therapy, Research Institute at Nationwide Children's Hospital & Ohio State University College of Medicine
Robert W Burgess, Jackson Laboratory & University of Maine

Document Type

Article

Publication Date

12-2-2019

Comments

This article has been peer-reviewed. It is the authors' final published version in the Journal of Clinical Investigation. Volume 129, Issue 12, December 2019, Pages 5568-5583.

The published version is available at https://doi.org/10.1172/JCI130600. Copyright © American Society for Clinical Investigation

Abstract

Gene therapy approaches are being deployed to treat recessive genetic disorders by restoring the expression of mutated genes. However, the feasibility of these approaches for dominantly inherited diseases - where treatment may require reduction in the expression of a toxic mutant protein resulting from a gain-of-function allele - is unclear. Here we show the efficacy of allele-specific RNAi as a potential therapy for Charcot-Marie-Tooth disease type 2D (CMT2D), caused by dominant mutations in glycyl-tRNA synthetase (GARS). A de novo mutation in GARS was identified in a patient with a severe peripheral neuropathy, and a mouse model precisely recreating the mutation was produced. These mice developed a neuropathy by 3-4 weeks of age, validating the pathogenicity of the mutation. RNAi sequences targeting mutant GARS mRNA, but not wild-type, were optimized and then packaged into AAV9 for in vivo delivery. This almost completely prevented the neuropathy in mice treated at birth. Delaying treatment until after disease onset showed modest benefit, though this effect decreased the longer treatment was delayed. These outcomes were reproduced in a second mouse model of CMT2D using a vector specifically targeting that allele. The effects were dose dependent, and persisted for at least 1 year. Our findings demonstrate the feasibility of AAV9-mediated allele-specific knockdown and provide proof of concept for gene therapy approaches for dominant neuromuscular diseases.

Recommended Citation

Morelli, Kathryn H; Griffin, Laurie B; Pyne, Nettie K; Wallace, Lindsay M; Fowler, Allison M; Oprescu, Stephanie N; Takase, Ryuichi; Wei, Na; Meyer-Schuman, Rebecca; Mellacheruvu, Dattatreya; Kitzman, Jacob O; Kocen, Samuel G; Hines, Timothy J; Spaulding, Emily L; Lupski, James R; Nesvizhskii, Alexey; Mancias, Pedro; Butler, Ian J; Yang, Xiang-Lei; Hou, Ya-Ming; Antonellis, Anthony; Harper, Scott Q; and Burgess, Robert W, "Allele-specific RNA interference prevents neuropathy in Charcot-Marie-Tooth disease type 2D mouse models." (2019). Department of Biochemistry and Molecular Biology Faculty Papers. Paper 163.
https://jdc.jefferson.edu/bmpfp/163

PubMed ID

31557132

Language

English