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ResearchIn-Press PreviewGeneticsOphthalmology Open Access | 10.1172/jci.insight.209108

Allotopic Expression of ND6 Restores Vision in a Mitochondrial Disease Model

Cheng Ai,1 Huiying Li,1 Jing Wu,1 Tianwei Zhou,1 Jing Wang,2 Shao-Hui Pan,3 Jun Yu,1 Douglas C. Wallace,4 and Min-Xin Guan1

1Center for Mitochondrial Biomedicine and Department of Ophthalmology, Fourth Affiliated Hospital of School of Medicine, Yiwu, China

2Institute of Genetics, Zhejiang University, Hangzhou, China

3Hangzhou Institute of Medicine, Wenzhou Medical University, Hangzhou, China

4Center for Mitochondrial and Epigenomic Medicine, Children's Hospital of Philadelphia, Philadelphia, United States of America

Find articles by Ai, C. in: PubMed | Google Scholar

1Center for Mitochondrial Biomedicine and Department of Ophthalmology, Fourth Affiliated Hospital of School of Medicine, Yiwu, China

2Institute of Genetics, Zhejiang University, Hangzhou, China

3Hangzhou Institute of Medicine, Wenzhou Medical University, Hangzhou, China

4Center for Mitochondrial and Epigenomic Medicine, Children's Hospital of Philadelphia, Philadelphia, United States of America

Find articles by Li, H. in: PubMed | Google Scholar |

1Center for Mitochondrial Biomedicine and Department of Ophthalmology, Fourth Affiliated Hospital of School of Medicine, Yiwu, China

2Institute of Genetics, Zhejiang University, Hangzhou, China

3Hangzhou Institute of Medicine, Wenzhou Medical University, Hangzhou, China

4Center for Mitochondrial and Epigenomic Medicine, Children's Hospital of Philadelphia, Philadelphia, United States of America

Find articles by Wu, J. in: PubMed | Google Scholar

1Center for Mitochondrial Biomedicine and Department of Ophthalmology, Fourth Affiliated Hospital of School of Medicine, Yiwu, China

2Institute of Genetics, Zhejiang University, Hangzhou, China

3Hangzhou Institute of Medicine, Wenzhou Medical University, Hangzhou, China

4Center for Mitochondrial and Epigenomic Medicine, Children's Hospital of Philadelphia, Philadelphia, United States of America

Find articles by Zhou, T. in: PubMed | Google Scholar

1Center for Mitochondrial Biomedicine and Department of Ophthalmology, Fourth Affiliated Hospital of School of Medicine, Yiwu, China

2Institute of Genetics, Zhejiang University, Hangzhou, China

3Hangzhou Institute of Medicine, Wenzhou Medical University, Hangzhou, China

4Center for Mitochondrial and Epigenomic Medicine, Children's Hospital of Philadelphia, Philadelphia, United States of America

Find articles by Wang, J. in: PubMed | Google Scholar

1Center for Mitochondrial Biomedicine and Department of Ophthalmology, Fourth Affiliated Hospital of School of Medicine, Yiwu, China

2Institute of Genetics, Zhejiang University, Hangzhou, China

3Hangzhou Institute of Medicine, Wenzhou Medical University, Hangzhou, China

4Center for Mitochondrial and Epigenomic Medicine, Children's Hospital of Philadelphia, Philadelphia, United States of America

Find articles by Pan, S. in: PubMed | Google Scholar

1Center for Mitochondrial Biomedicine and Department of Ophthalmology, Fourth Affiliated Hospital of School of Medicine, Yiwu, China

2Institute of Genetics, Zhejiang University, Hangzhou, China

3Hangzhou Institute of Medicine, Wenzhou Medical University, Hangzhou, China

4Center for Mitochondrial and Epigenomic Medicine, Children's Hospital of Philadelphia, Philadelphia, United States of America

Find articles by Yu, J. in: PubMed | Google Scholar

1Center for Mitochondrial Biomedicine and Department of Ophthalmology, Fourth Affiliated Hospital of School of Medicine, Yiwu, China

2Institute of Genetics, Zhejiang University, Hangzhou, China

3Hangzhou Institute of Medicine, Wenzhou Medical University, Hangzhou, China

4Center for Mitochondrial and Epigenomic Medicine, Children's Hospital of Philadelphia, Philadelphia, United States of America

Find articles by Wallace, D. in: PubMed | Google Scholar

1Center for Mitochondrial Biomedicine and Department of Ophthalmology, Fourth Affiliated Hospital of School of Medicine, Yiwu, China

2Institute of Genetics, Zhejiang University, Hangzhou, China

3Hangzhou Institute of Medicine, Wenzhou Medical University, Hangzhou, China

4Center for Mitochondrial and Epigenomic Medicine, Children's Hospital of Philadelphia, Philadelphia, United States of America

Find articles by Guan, M. in: PubMed | Google Scholar |

Published September 8, 2026 - More info

JCI Insight. https://doi.org/10.1172/jci.insight.209108.
Copyright © 2026, Ai et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Published September 8, 2026 - Version history
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Abstract

Mutations in mitochondrial DNA (mtDNA) cause various mitochondrial diseases that are currently incurable. Allotopic expression of nuclear-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 pre-symptomatic 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 provide critical mechanistic insights into its role in treating LHON and other mitochondrial diseases.

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