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Updated: Jan 8, 2026
Adeno-Associated Virus-Mediated Delivery of CRISPR for Cardiac Gene Editing in Mice
Published on: August 2, 2018
Partial restoration of mitochondrial dysfunction by AAV-Ant1 protects from dilated cardiomyopathy in Ant1-/- plus
Alessia Angelin1,2, Kierstin Keller1, Peiran Lu1
1Center for Mitochondrial and Epigenomic Medicine, The Children's Hospital of Philadelphia, Philadelphia, PA, USA.
Abstract:
Primary mitochondrial disease (PMD) patients manifesting cardiomyopathy are twice as likely to die as other PMD patients. One PMD with cardiomyopathy is caused by null mutations in the heart-muscle isoform of the adenine nucleotide translocator (SLC25A4, ANT1) gene, with the severity of cardiomyopathy mediated by mitochondrial DNA. To optimize strategies for addressing mitochondrial cardiomyopathy, we generated an Ant1 null mouse and combined it with the ND6P25L mitochondrial DNA mutation to mimic the hypertrophic versus dilated cardiomyopathies observed in patients. Here, we transduce the neonatal Ant1-/- and Ant1-/-+ND6P25L mouse hearts with an AAV2/9-pDes-Gfp-mAnt1 cDNA vector. We show that restoration of just 10% of Ant1 gene expression was sufficient to ameliorate the cardiomyopathies in these mice. Proteomics and single-nucleus RNA sequencing reveal the reversal of dysregulated mitochondrial metabolic genes, including PGC1α, as well as cardiac contractile and extracellular matrix proteins. Hence, a modest increase in cardiac mitochondrial energetics can have profound benefits on cardiac function and is effective in treating mitochondrial cardiomyopathy.
Insights
Restoring even 10% of adenine nucleotide translocator (ANT1) gene expression ameliorated mitochondrial cardiomyopathy in mice. This finding offers a promising therapeutic strategy for heart conditions caused by mitochondrial dysfunction.
Area of Science:
- Cardiology
- Genetics
- Mitochondrial Biology
Background:
- Primary mitochondrial diseases (PMD) with cardiomyopathy significantly increase mortality risk.
- Adenine nucleotide translocator (ANT1) gene mutations are a known cause of PMD cardiomyopathy, with severity influenced by mitochondrial DNA.
- Existing mouse models do not fully recapitulate the spectrum of human mitochondrial cardiomyopathy.
Purpose of the Study:
- To develop and validate a mouse model for studying ANT1-related mitochondrial cardiomyopathy.
- To investigate the therapeutic potential of restoring ANT1 gene expression in affected hearts.
- To elucidate the molecular mechanisms underlying therapeutic benefits.
Main Methods:
- Generation of Ant1 null mice, combined with a specific mitochondrial DNA mutation (ND6P25L) to model human cardiomyopathies.
- Cardiac gene therapy using an adeno-associated virus vector (AAV2/9-pDes-Gfp-mAnt1 cDNA) to restore ANT1 expression in neonatal mice.
- Proteomic and single-nucleus RNA sequencing analyses to assess molecular changes post-treatment.
Main Results:
- Restoration of approximately 10% of cardiac ANT1 gene expression was sufficient to significantly improve cardiomyopathy in both Ant1 null and Ant1-/-+ND6P25L mice.
- Therapeutic intervention reversed dysregulation in key mitochondrial metabolic genes, including PGC1α.
- Cardiac contractile and extracellular matrix protein profiles were normalized, indicating functional recovery.
Conclusions:
- Modest enhancement of cardiac mitochondrial energetics can profoundly benefit heart function in mitochondrial cardiomyopathy.
- Gene therapy targeting ANT1 represents a viable and effective strategy for treating this debilitating condition.
- This study provides a foundation for developing targeted therapies for mitochondrial cardiomyopathies.
