Clinically translatable mitochondrial gene therapy in muscle using tandem mtZFN architecture

Abstract Mutations in the mitochondrial genome (mtDNA) often lead to clinical pathologies. Mitochondrially-targeted zinc finger nucleases (mtZFNs) have been successful in reducing the levels of mutation-bearing mtDNA both in vivo and in vitro, resulting in a shift in the genetic makeup of affected m...

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Main Authors: Pavel A Nash, Keira M Turner, Christopher A Powell, Lindsey Van Haute, Pedro Silva-Pinheiro, Felix Bubeck, Ellen Wiedtke, Eloïse Marques, Dylan G Ryan, Dirk Grimm, Payam A Gammage, Michal Minczuk
Format: Article
Language:English
Published: Springer Nature 2025-04-01
Series:EMBO Molecular Medicine
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Online Access:https://doi.org/10.1038/s44321-025-00231-5
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author Pavel A Nash
Keira M Turner
Christopher A Powell
Lindsey Van Haute
Pedro Silva-Pinheiro
Felix Bubeck
Ellen Wiedtke
Eloïse Marques
Dylan G Ryan
Dirk Grimm
Payam A Gammage
Michal Minczuk
author_facet Pavel A Nash
Keira M Turner
Christopher A Powell
Lindsey Van Haute
Pedro Silva-Pinheiro
Felix Bubeck
Ellen Wiedtke
Eloïse Marques
Dylan G Ryan
Dirk Grimm
Payam A Gammage
Michal Minczuk
author_sort Pavel A Nash
collection DOAJ
description Abstract Mutations in the mitochondrial genome (mtDNA) often lead to clinical pathologies. Mitochondrially-targeted zinc finger nucleases (mtZFNs) have been successful in reducing the levels of mutation-bearing mtDNA both in vivo and in vitro, resulting in a shift in the genetic makeup of affected mitochondria and subsequently to phenotypic rescue. Given the uneven distribution in the mtDNA mutation load across tissues in patients, and a great diversity in pathogenic mutations, it is of interest to develop mutation-specific, selective gene therapies that could be delivered to particular tissues. This study demonstrates the effectiveness of in vivo mitochondrial gene therapy using a novel mtZFN architecture on skeletal muscle using adeno-associated viral (AAV) platforms in a murine model harboring a pathogenic mtDNA mutation. We observed effective reduction in mutation load of cardiac and skeletal muscle, which was accompanied by molecular phenotypic rescue. The gene therapy treatment was shown to be safe when markers of immunity and inflammation were assessed. These results highlight the potential of curative approaches for mitochondrial diseases, paving the way for targeted and effective treatments.
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spelling doaj-art-0ea527046bec41c6b2e7dd9f4a54ca992025-08-20T02:06:35ZengSpringer NatureEMBO Molecular Medicine1757-46842025-04-011761222123710.1038/s44321-025-00231-5Clinically translatable mitochondrial gene therapy in muscle using tandem mtZFN architecturePavel A Nash0Keira M Turner1Christopher A Powell2Lindsey Van Haute3Pedro Silva-Pinheiro4Felix Bubeck5Ellen Wiedtke6Eloïse Marques7Dylan G Ryan8Dirk Grimm9Payam A Gammage10Michal Minczuk11MRC Mitochondrial Biology Unit, University of CambridgeMRC Mitochondrial Biology Unit, University of CambridgeMRC Mitochondrial Biology Unit, University of CambridgeMRC Mitochondrial Biology Unit, University of CambridgeMRC Mitochondrial Biology Unit, University of CambridgeDepartment of Infectious Diseases/Virology, Section Viral Vector Technologies, Medical Faculty, Heidelberg UniversityDepartment of Infectious Diseases/Virology, Section Viral Vector Technologies, Medical Faculty, Heidelberg UniversityMRC Mitochondrial Biology Unit, University of CambridgeMRC Mitochondrial Biology Unit, University of CambridgeDepartment of Infectious Diseases/Virology, Section Viral Vector Technologies, Medical Faculty, Heidelberg UniversityMRC Mitochondrial Biology Unit, University of CambridgeMRC Mitochondrial Biology Unit, University of CambridgeAbstract Mutations in the mitochondrial genome (mtDNA) often lead to clinical pathologies. Mitochondrially-targeted zinc finger nucleases (mtZFNs) have been successful in reducing the levels of mutation-bearing mtDNA both in vivo and in vitro, resulting in a shift in the genetic makeup of affected mitochondria and subsequently to phenotypic rescue. Given the uneven distribution in the mtDNA mutation load across tissues in patients, and a great diversity in pathogenic mutations, it is of interest to develop mutation-specific, selective gene therapies that could be delivered to particular tissues. This study demonstrates the effectiveness of in vivo mitochondrial gene therapy using a novel mtZFN architecture on skeletal muscle using adeno-associated viral (AAV) platforms in a murine model harboring a pathogenic mtDNA mutation. We observed effective reduction in mutation load of cardiac and skeletal muscle, which was accompanied by molecular phenotypic rescue. The gene therapy treatment was shown to be safe when markers of immunity and inflammation were assessed. These results highlight the potential of curative approaches for mitochondrial diseases, paving the way for targeted and effective treatments.https://doi.org/10.1038/s44321-025-00231-5Gene TherapyZinc Finger Nuclease (mtZFN)Adeno-Associated Viruses (AAV)mtDNA Heteroplasmy ModificationSkeletal Muscle
spellingShingle Pavel A Nash
Keira M Turner
Christopher A Powell
Lindsey Van Haute
Pedro Silva-Pinheiro
Felix Bubeck
Ellen Wiedtke
Eloïse Marques
Dylan G Ryan
Dirk Grimm
Payam A Gammage
Michal Minczuk
Clinically translatable mitochondrial gene therapy in muscle using tandem mtZFN architecture
EMBO Molecular Medicine
Gene Therapy
Zinc Finger Nuclease (mtZFN)
Adeno-Associated Viruses (AAV)
mtDNA Heteroplasmy Modification
Skeletal Muscle
title Clinically translatable mitochondrial gene therapy in muscle using tandem mtZFN architecture
title_full Clinically translatable mitochondrial gene therapy in muscle using tandem mtZFN architecture
title_fullStr Clinically translatable mitochondrial gene therapy in muscle using tandem mtZFN architecture
title_full_unstemmed Clinically translatable mitochondrial gene therapy in muscle using tandem mtZFN architecture
title_short Clinically translatable mitochondrial gene therapy in muscle using tandem mtZFN architecture
title_sort clinically translatable mitochondrial gene therapy in muscle using tandem mtzfn architecture
topic Gene Therapy
Zinc Finger Nuclease (mtZFN)
Adeno-Associated Viruses (AAV)
mtDNA Heteroplasmy Modification
Skeletal Muscle
url https://doi.org/10.1038/s44321-025-00231-5
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