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.

Public Health Relevance

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.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY010804-18
Application #
8589139
Study Section
Therapeutic Approaches to Genetic Diseases (TAG)
Program Officer
Araj, Houmam H
Project Start
1994-12-01
Project End
2016-08-31
Budget Start
2013-09-30
Budget End
2014-08-31
Support Year
18
Fiscal Year
2013
Total Cost
$383,750
Indirect Cost
$133,750
Name
University of Miami School of Medicine
Department
Neurology
Type
Schools of Medicine
DUNS #
052780918
City
Coral Gables
State
FL
Country
United States
Zip Code
33146
Wang, Xiao; Peralta, Susana; Moraes, Carlos T (2013) Mitochondrial alterations during carcinogenesis: a review of metabolic transformation and targets for anticancer treatments. Adv Cancer Res 119:127-60
Pickrell, Alicia M; Pinto, Milena; Moraes, Carlos T (2013) Mouse models of Parkinson's disease associated with mitochondrial dysfunction. Mol Cell Neurosci 55:87-94
Pinto, Milena; Pickrell, Alicia M; Fukui, Hirokazu et al. (2013) Mitochondrial DNA damage in a mouse model of Alzheimer's disease decreases amyloid beta plaque formation. Neurobiol Aging 34:2399-407
Moraes, Carlos T (2013) Adrenoleukodystrophy and the mitochondrial connection: clues for supplementing Lorenzo's oil. Brain 136:2339-41
Bacman, Sandra R; Williams, Sion L; Pinto, Milena et al. (2013) Specific elimination of mutant mitochondrial genomes in patient-derived cells by mitoTALENs. Nat Med 19:1111-3
Wang, Xiao; Pickrell, Alicia M; Rossi, Susana G et al. (2013) Transient systemic mtDNA damage leads to muscle wasting by reducing the satellite cell pool. Hum Mol Genet 22:3976-86
Diaz, Francisca; Enriquez, Jose Antonio; Moraes, Carlos T (2012) Cells lacking Rieske iron-sulfur protein have a reactive oxygen species-associated decrease in respiratory complexes I and IV. Mol Cell Biol 32:415-29
Dillon, Lloye M; Williams, Sion L; Hida, Aline et al. (2012) Increased mitochondrial biogenesis in muscle improves aging phenotypes in the mtDNA mutator mouse. Hum Mol Genet 21:2288-97
Dillon, Lloye M; Rebelo, Adriana P; Moraes, Carlos T (2012) The role of PGC-1 coactivators in aging skeletal muscle and heart. IUBMB Life 64:231-41
Wenz, Tina; Wang, Xiao; Marini, Matteo et al. (2011) A metabolic shift induced by a PPAR panagonist markedly reduces the effects of pathogenic mitochondrial tRNA mutations. J Cell Mol Med 15:2317-25

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