AAV-delivered suppressor tRNA overcomes a nonsense mutation in mice
Authors
Wang, JiamingZhang, Yue
Mendonca, Craig A.
Yukselen, Onur
Muneeruddin, Khaja
Ren, Lingzhi
Liang, Jialing
Zhou, Chen
Xie, Jun
Li, Jia
Jiang, Zhong
Kucukural, Alper
Shaffer, Scott A.
Gao, Guangping
Wang, Dan
UMass Chan Affiliations
RNA Therapeutics InstituteDepartment of Pathology
Department of Microbiology and Physiological Systems
Department of Biochemistry and Molecular Pharmacology
Mass Spectrometry Facility
Bioinformatics Core
Program in Molecular Medicine
Horae Gene Therapy Center
Document Type
Journal ArticlePublication Date
2022-03-23
Metadata
Show full item recordAbstract
Gene therapy is a potentially curative medicine for many currently untreatable diseases, and recombinant adeno-associated virus (rAAV) is the most successful gene delivery vehicle for in vivo applications(1-3). However, rAAV-based gene therapy suffers from several limitations, such as constrained DNA cargo size and toxicities caused by non-physiological expression of a transgene(4-6). Here we show that rAAV delivery of a suppressor tRNA (rAAV.sup-tRNA) safely and efficiently rescued a genetic disease in a mouse model carrying a nonsense mutation, and effects lasted for more than 6 months after a single treatment. Mechanistically, this was achieved through a synergistic effect of premature stop codon readthrough and inhibition of nonsense-mediated mRNA decay. rAAV.sup-tRNA had a limited effect on global readthrough at normal stop codons and did not perturb endogenous tRNA homeostasis, as determined by ribosome profiling and tRNA sequencing, respectively. By optimizing the AAV capsid and the route of administration, therapeutic efficacy in various target tissues was achieved, including liver, heart, skeletal muscle and brain. This study demonstrates the feasibility of developing a toolbox of AAV-delivered nonsense suppressor tRNAs operating on premature termination codons (AAV-NoSTOP) to rescue pathogenic nonsense mutations and restore gene function under endogenous regulation. As nonsense mutations account for 11% of pathogenic mutations, AAV-NoSTOP can benefit a large number of patients. AAV-NoSTOP obviates the need to deliver a full-length protein-coding gene that may exceed the rAAV packaging limit, elicit adverse immune responses or cause transgene-related toxicities. It therefore represents a valuable addition to gene therapeutics.Source
Wang J, Zhang Y, Mendonca CA, Yukselen O, Muneeruddin K, Ren L, Liang J, Zhou C, Xie J, Li J, Jiang Z, Kucukural A, Shaffer SA, Gao G, Wang D. AAV-delivered suppressor tRNA overcomes a nonsense mutation in mice. Nature. 2022 Apr;604(7905):343-348. doi: 10.1038/s41586-022-04533-3. Epub 2022 Mar 23. PMID: 35322228. Link to article on publisher's site
DOI
10.1038/s41586-022-04533-3Permanent Link to this Item
http://hdl.handle.net/20.500.14038/25964PubMed ID
35322228Related Resources
ae974a485f413a2113503eed53cd6c53
10.1038/s41586-022-04533-3