Our long-term goal is to develop genetic approaches that could be used for the treatment of mitochondrial disorders associated with mitochondrial DNA (mtDNA) mutations. We propose to develop methods that improve oxidative phosphorylation function in cells harboring heteroplasmic mtDNA mutations (i.e. a mixture of wild-type and mutated mtDNA). Our approach is based on the "mtDNA heteroplasmy shift" concept, where, by genetic manipulation, the levels of the mutated genome are decreased in relation to the wild-type genome, thereby improving cellular health. In the previous funding period we showed that mitochondria-targeted restriction endonucleases are powerful tools to achieve mtDNA heteroplasmy shift. The present proposal will further expand this approach to any mtDNA mutation. To do so, we will take advantage of new developments in the field of designed nucleases. TAL-effector nucleases have emerged as highly flexible molecules that can recognize DNA regions based on a modular arrangement of their DNA binding domains. We have obtained preliminary data showing that this approach is feasible and propose to better characterize the system in vitro, ex vivo and in vivo.
We propose to use mitochondrial-targeted designer nucleases to eliminate the mutated mitochondrial genome in cells harboring a mixture of mutated and wild-type mitochondrial DNA. This change would lead to a correction of biochemical defects with a potential curative outcome in certain patients.
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|Luo, Xueting; Ribeiro, Marcio; Bray, Eric R et al. (2016) Enhanced Transcriptional Activity and Mitochondrial Localization of STAT3 Co-induce Axon Regrowth in the Adult Central Nervous System. Cell Rep 15:398-410|
|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|
|Pinto, Milena; Moraes, Carlos T (2015) Mechanisms linking mtDNA damage and aging. Free Radic Biol Med 85:250-8|
|(2015) Retraction Notice to: mTERF2 Regulates Oxidative Phosphorylation by Modulating mtDNA Transcription. Cell Metab 22:751|
|Hashimoto, Masami; Bacman, Sandra R; Peralta, Susana et al. (2015) MitoTALEN: A General Approach to Reduce Mutant mtDNA Loads and Restore Oxidative Phosphorylation Function in Mitochondrial Diseases. Mol Ther 23:1592-9|
|Reddy, Pradeep; Ocampo, Alejandro; Suzuki, Keiichiro et al. (2015) Selective elimination of mitochondrial mutations in the germline by genome editing. Cell 161:459-69|
|Pinto, Milena; Moraes, Carlos T (2014) Mitochondrial genome changes and neurodegenerative diseases. Biochim Biophys Acta 1842:1198-207|
|Bacman, Sandra R; Williams, Sion L; Pinto, Milena et al. (2014) The use of mitochondria-targeted endonucleases to manipulate mtDNA. Methods Enzymol 547:373-97|
|Moraes, Carlos T; Bacman, Sandra R; Williams, Sion L (2014) Manipulating mitochondrial genomes in the clinic: playing by different rules. Trends Cell Biol 24:209-11|
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