TY - JOUR
T1 - Targeted genome editing restores auditory function in adult mice with progressive hearing loss caused by a human microRNA mutation
AU - Zhu, Wenliang
AU - Du, Wan
AU - Rameshbabu, Arun Prabhu
AU - Armstrong, Ariel Miura
AU - Silver, Stewart
AU - Kim, Yehree
AU - Wei, Wei
AU - Shu, Yilai
AU - Liu, Xuezhong
AU - Lewis, Morag A.
AU - Steel, Karen P.
AU - Chen, Zheng Yi
N1 - Publisher Copyright:
Copyright © 2024 The Authors, some rights reserved
PY - 2024/7/10
Y1 - 2024/7/10
N2 - Mutations in microRNA-96 (MIR96) cause autosomal dominant deafness-50 (DFNA50), a form of delayed-onset hearing loss. Genome editing has shown efficacy in hearing recovery through intervention in neonatal mice, yet editing in the adult inner ear is necessary for clinical applications, which has not been done. Here, we developed a genome editing therapy for the MIR96 mutation 14C>A by screening different CRISPR systems and optimizing Cas9 expression and the sgRNA scaffold for efficient and specific mutation editing. AAV delivery of the KKH variant of Staphylococcus aureus Cas9 (SaCas9-KKH) and sgRNA to the cochleae of presymptomatic (3-week-old) and symptomatic (6-week-old) adult Mir9614C>A/+ mutant mice improved hearing long term, with efficacy increased by injection at a younger age. Adult inner ear delivery resulted in transient Cas9 expression without evidence of AAV genomic integration, indicating the good safety profile of our in vivo genome editing strategy. We developed a dual-AAV system, including an AAV-sgmiR96-master carrying sgRNAs against all known human MIR96 mutations. Because mouse and human MIR96 sequences share 100% homology, our approach and sgRNA selection for efficient and specific hair cell editing for long-term hearing recovery lay the foundation for the development of treatment for patients with DFNA50 caused by MIR96 mutations.
AB - Mutations in microRNA-96 (MIR96) cause autosomal dominant deafness-50 (DFNA50), a form of delayed-onset hearing loss. Genome editing has shown efficacy in hearing recovery through intervention in neonatal mice, yet editing in the adult inner ear is necessary for clinical applications, which has not been done. Here, we developed a genome editing therapy for the MIR96 mutation 14C>A by screening different CRISPR systems and optimizing Cas9 expression and the sgRNA scaffold for efficient and specific mutation editing. AAV delivery of the KKH variant of Staphylococcus aureus Cas9 (SaCas9-KKH) and sgRNA to the cochleae of presymptomatic (3-week-old) and symptomatic (6-week-old) adult Mir9614C>A/+ mutant mice improved hearing long term, with efficacy increased by injection at a younger age. Adult inner ear delivery resulted in transient Cas9 expression without evidence of AAV genomic integration, indicating the good safety profile of our in vivo genome editing strategy. We developed a dual-AAV system, including an AAV-sgmiR96-master carrying sgRNAs against all known human MIR96 mutations. Because mouse and human MIR96 sequences share 100% homology, our approach and sgRNA selection for efficient and specific hair cell editing for long-term hearing recovery lay the foundation for the development of treatment for patients with DFNA50 caused by MIR96 mutations.
UR - http://www.scopus.com/inward/record.url?scp=85198460539&partnerID=8YFLogxK
U2 - 10.1126/scitranslmed.adn0689
DO - 10.1126/scitranslmed.adn0689
M3 - Article
C2 - 38985856
AN - SCOPUS:85198460539
SN - 1946-6234
VL - 16
JO - Science translational medicine
JF - Science translational medicine
IS - 755
M1 - eadn0689
ER -
