Our long term goal is to develop genetic approaches that eventually could be used for the treatment of Mitochondrial Disorders associated with mitochondrial DNA (mtDNA) mutations. Although some of the approaches tested by us and others in the past showed some promise, they are inefficient and limited to correcting problems in a few genes. We now propose to develop approaches that are more efficient in improving oxidative phosphorylation in cells harboring heteroplasmic mtDNA mutations (i.e. a mixture of wild-type and mutated mtDNA). The approach is based on the """"""""mtDNA heteroplasmy shift"""""""" concept, where, by genetic manipulation, levels of the wild-type genome are increased in relation to the mutated genome. Our proposal will apply two related but different tools to induce mtDNA heteroplasmy shift. The first one is the use mitochondrially-targeted restriction endonucleases that can specifically cleave mitochondrial genomes containing mutated sites. We have shown that such approach can shift mtDNA heteroplasmy efficiently in cultured cells. We now propose to continue these studies in cultured cells and to test the efficiency and safety of the procedure in transgenic mice. Because restriction endonucleases are also limited by the number of recognition sites, we propose to expand this approach by using a second tool, namely mitochondrially-targeted chimeric Zinc Finger Proteins and type IIS restriction endonucleases' catalytic domain. Zinc Finger Proteins can be designed and fined tuned for the recognition of novel DNA sites. Once the DNA binding specificity is obtained, the addition of the catalytic domain from type IIS restriction endonucleases (Fokl) to the Zinc Finger DNA binding region would create molecules with a potential to shift mtDNA heteroplasmy of mutations that are prevalent in the patient population.

National Institute of Health (NIH)
National Eye Institute (NEI)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-SSS-G (03))
Program Officer
Chin, Hemin R
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Miami School of Medicine
Schools of Medicine
Coral Gables
United States
Zip Code
Nissanka, Nadee; Moraes, Carlos T (2018) Mitochondrial DNA damage and reactive oxygen species in neurodegenerative disease. FEBS Lett 592:728-742
Pinto, Milena; Nissanka, Nadee; Moraes, Carlos T (2018) Lack of Parkin Anticipates the Phenotype and Affects Mitochondrial Morphology and mtDNA Levels in a Mouse Model of Parkinson's Disease. J Neurosci 38:1042-1053
Peralta, Susana; Goffart, Steffi; Williams, Sion L et al. (2018) ATAD3 controls mitochondrial cristae structure in mouse muscle, influencing mtDNA replication and cholesterol levels. J Cell Sci 131:
Garcia, Sofia; Nissanka, Nadee; Mareco, Edson A et al. (2018) Overexpression of PGC-1? in aging muscle enhances a subset of young-like molecular patterns. Aging Cell 17:
Arguello, Tania; Köhrer, Caroline; RajBhandary, Uttam L et al. (2018) Mitochondrial methionyl N-formylation affects steady-state levels of oxidative phosphorylation complexes and their organization into supercomplexes. J Biol Chem 293:15021-15032
Madsen, Pernille M; Pinto, Milena; Patel, Shreyans et al. (2017) Mitochondrial DNA Double-Strand Breaks in Oligodendrocytes Cause Demyelination, Axonal Injury, and CNS Inflammation. J Neurosci 37:10185-10199
Pinto, Milena; Pickrell, Alicia M; Wang, Xiao et al. (2017) Transient mitochondrial DNA double strand breaks in mice cause accelerated aging phenotypes in a ROS-dependent but p53/p21-independent manner. Cell Death Differ 24:288-299
Tengan, Celia H; Moraes, Carlos T (2017) NO control of mitochondrial function in normal and transformed cells. Biochim Biophys Acta Bioenerg 1858:573-581
Peralta, Susana; Garcia, Sofia; Yin, Han Yang et al. (2016) Sustained AMPK activation improves muscle function in a mitochondrial myopathy mouse model by promoting muscle fiber regeneration. Hum Mol Genet 25:3178-3191
Pinto, Milena; Nissanka, Nadee; Peralta, Susana et al. (2016) Pioglitazone ameliorates the phenotype of a novel Parkinson's disease mouse model by reducing neuroinflammation. Mol Neurodegener 11:25

Showing the most recent 10 out of 88 publications