Allotopic expression of ND6 restores vision in a mitochondrial disease model
5 affiliations
- 1Center for Mitochondrial Biomedicine and Department of Ophthalmology, the Fourth Affiliated Hospital of School of Medicine, Zhejiang University, Yiwu, Zhejiang, China.
- 2Center for Genetic Medicine, International School of Medicine and International Institute of Medicine, and
- 3Institute of Genetics, Zhejiang University, Hangzhou, Zhejiang, China.
- 4Eye Research Center, Hangzhou Institute of Medicine, Chinese Academy of Sciences, Eye Hospital, Wenzhou Medical University, Hangzhou, China.
- 5Center for Mitochondrial and Epigenomic Medicine, Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania, USA.
Abstract
Mutations in mitochondrial DNA (mtDNA) cause various mitochondrial diseases that are currently incurable. Allotopic expression of nucleus-recoded mitochondrial genes represents a promising therapeutic strategy, given its demonstrated capacity to restore mitochondrial function in human cell models harboring mtDNA mutations. However, the in vivo evaluation of allotopic gene therapy has been hindered by optimization challenges and the lack of appropriate animal models. Here, we overcome these limitations by utilizing an optimized AAV2-ND6 construct with codon optimization and mitochondrial targeting sequence in a mouse model bearing the homoplasmic ND6P25L mutation, which recapitulates Leber hereditary optic neuropathy (LHON). High-dose administration of the AAV2-ND6 construct resulted in robust, sustained expression within the retina and optic nerve without apparent systemic toxicity. Strikingly, we compared the therapeutic efficacy in mutant mice at different ages and presymptomatic intervention with AAV2-ND6 effectively attenuated disease progression, mitigated retinal cellular deficiencies and optic nerve damage, and restored visual function in ND6P25L mice. Mechanistically, allotopic ND6 expression markedly rescued the mitochondrial dysfunction, corrected dysregulated retinol metabolism and phototransduction pathways, and suppressed apoptotic processes in the mutant retina. Our study validates the safety and therapeutic potential of allotopic expression in vivo and provides critical mechanistic insights into its role in treating LHON and other mitochondrial diseases.
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BibTeX
@article{ai2026allotopic,
title = {{Allotopic expression of ND6 restores vision in a mitochondrial disease model}},
author = {Cheng Ai and Huiying Li and Jing Wu and Tianwei Zhou and Jing Wang and Shao-Hui Pan and Jun Yu and Douglas C. Wallace and Min-Xin Guan},
journal = {JCI Insight},
year = {2026},
month = sep,
doi = {10.1172/jci.insight.209108},
url = {https://doi.org/10.1172/jci.insight.209108},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Allotopic expression of ND6 restores vision in a mitochondrial disease model
AU - Cheng Ai
AU - Huiying Li
AU - Jing Wu
AU - Tianwei Zhou
AU - Jing Wang
AU - Shao-Hui Pan
AU - Jun Yu
AU - Douglas C. Wallace
AU - Min-Xin Guan
PY - 2026
DA - 2026/09/08
JO - JCI Insight
DO - 10.1172/jci.insight.209108
UR - https://doi.org/10.1172/jci.insight.209108
AB - Mutations in mitochondrial DNA (mtDNA) cause various mitochondrial diseases that are currently incurable. Allotopic expression of nucleus-recoded mitochondrial genes represents a promising therapeutic strategy, given its demonstrated capacity to restore mitochondrial function in human cell models harboring mtDNA mutations. However, the in vivo evaluation of allotopic gene therapy has been hindered by optimization challenges and the lack of appropriate animal models. Here, we overcome these limitations by utilizing an optimized AAV2-ND6 construct with codon optimization and mitochondrial targeting sequence in a mouse model bearing the homoplasmic ND6P25L mutation, which recapitulates Leber hereditary optic neuropathy (LHON). High-dose administration of the AAV2-ND6 construct resulted in robust, sustained expression within the retina and optic nerve without apparent systemic toxicity. Strikingly, we compared the therapeutic efficacy in mutant mice at different ages and presymptomatic intervention with AAV2-ND6 effectively attenuated disease progression, mitigated retinal cellular deficiencies and optic nerve damage, and restored visual function in ND6P25L mice. Mechanistically, allotopic ND6 expression markedly rescued the mitochondrial dysfunction, corrected dysregulated retinol metabolism and phototransduction pathways, and suppressed apoptotic processes in the mutant retina. Our study validates the safety and therapeutic potential of allotopic expression in vivo and provides critical mechanistic insights into its role in treating LHON and other mitochondrial diseases.
SN - 2379-3708
ER -