Mitochondrial Gene Therapy Augments Mitochondrial Physiology in a Parkinson's Disease Cell Model
| Autor: | Kristen E. Bergquist, Kathleen M. Schwarz, Patricia A. Trimmer, Shilpa Iyer, Ravindar R. Thomas, Christina M. Papageorge, James P. Bennett, Shaharyar M. Khan, Lisa D. Dunham, Caitlin K. Quigley, Paula M. Keeney, Francisco R. Portell |
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| Rok vydání: | 2009 |
| Předmět: |
Scientific Articles
Mitochondrial DNA Cell Respiration SOD2 Substantia nigra Protein Sorting Signals Mitochondrion Biology DNA Mitochondrial Human mitochondrial genetics Mitochondrial Proteins Cell Line Tumor Genetics medicine Humans Molecular Biology Neurodegeneration Parkinson Disease Genetic Therapy TFAM medicine.disease Mitochondria Protein Structure Tertiary Cell biology DNA-Binding Proteins Genes Mitochondrial Gene Expression Regulation Small Ubiquitin-Related Modifier Proteins Molecular Medicine Transcription Factors Protofection |
| Zdroj: | Human Gene Therapy. 20:897-907 |
| ISSN: | 1557-7422 1043-0342 |
| DOI: | 10.1089/hum.2009.023 |
| Popis: | Neurodegeneration in Parkinson's disease (PD) affects mainly dopaminergic neurons in the substantia nigra, where age-related, increasing percentages of cells lose detectable respiratory activity associated with depletion of intact mitochondrial DNA (mtDNA). Replenishment of mtDNA might improve neuronal bioenergetic function and prevent further cell death. We developed a technology ("ProtoFection") that uses recombinant human mitochondrial transcription factor A (TFAM) engineered with an N-terminal protein transduction domain (PTD) followed by the SOD2 mitochondrial localization signal (MLS) to deliver mtDNA cargo to the mitochondria of living cells. MTD-TFAM (MTD = PTD + MLS = "mitochondrial transduction domain") binds mtDNA and rapidly transports it across plasma membranes to mitochondria. For therapeutic proof-of-principle we tested ProtoFection technology in Parkinson's disease cybrid cells, using mtDNA generated from commercially available human genomic DNA (gDNA; Roche). Nine to 11 weeks after single exposures to MTD-TFAM + mtDNA complex, PD cybrid cells with impaired respiration and reduced mtDNA genes increased their mtDNA gene copy numbers up to 24-fold, mtDNA-derived RNAs up to 35-fold, TFAM and ETC proteins, cell respiration, and mitochondrial movement velocities. Cybrid cells with no or minimal basal mitochondrial impairments showed reduced or no responses to treatment, suggesting the possibility of therapeutic selectivity. Exposure of PD but not control cybrid cells to MTD-TFAM protein alone or MTD-TFAM + mtDNA complex increased expression of PGC-1alpha, suggesting activation of mitochondrial biogenesis. ProtoFection technology for mitochondrial gene therapy holds promise for improving bioenergetic function in impaired PD neurons and needs additional development to define its pharmacodynamics and delineate its molecular mechanisms. It also is unclear whether single-donor gDNA for generating mtDNA would be a preferred therapeutic compared with the pooled gDNA used in this study. |
| Databáze: | OpenAIRE |
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