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Difference between revisions of "Keeney 2009 Hum Gene Ther"

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{{Publication
{{Publication
|title=Keeney PM, Quigley CK, Dunham LD, Papageorge CM, Iyer S, Thomas RR, Schwarz KM, Trimmer PA, Khan SM, Portell FR, Bergquist KE, Bennett JP (2009) Mitochondrial gene therapy augments mitochondrial physiology in a Parkinson's disease cell model. Hum Gene Ther 20: 897-907.
|title=Keeney PM, Quigley CK, Dunham LD, Papageorge CM, Iyer S, Thomas RR, Schwarz KM, Trimmer PA, Khan SM, Portell FR, Bergquist KE, Bennett JP (2009) Mitochondrial gene therapy augments mitochondrial physiology in a Parkinson's disease cell model. Hum Gene Ther 20:897-907.
|info=[http://www.ncbi.nlm.nih.gov/pubmed/19374590 PMID: 19374590 Open Access]
|info=[http://www.ncbi.nlm.nih.gov/pubmed/19374590 PMID: 19374590 Open Access]
|authors=Keeney PM, Quigley CK, Dunham LD, Papageorge CM, Iyer S, Thomas RR, Schwarz KM, Trimmer PA, Khan SM, Portell FR, Bergquist KE, Bennett JP
|authors=Keeney PM, Quigley CK, Dunham LD, Papageorge CM, Iyer S, Thomas RR, Schwarz KM, Trimmer PA, Khan SM, Portell FR, Bergquist KE, Bennett JP
Line 6: Line 6:
|journal=Hum Gene Ther
|journal=Hum Gene Ther
|abstract=Neurodegeneration in Parkinsonā€™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 (ā€˜ā€˜ProtoFectionā€™ā€™) 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. MTDā€“TFAM (MTDĀ¼PTDĆ¾MLSĀ¼ā€˜ā€˜mitochondrial transduction domainā€™ā€™) binds mtDNA and rapidly transports it across plasma membranes to mitochondria. For therapeutic proof-of-principle we tested ProtoFection technology in Parkinsonā€™s disease cybrid cells, using mtDNA generated from commercially available human genomic DNA (gDNA; Roche). Nine to 11 weeks after single exposures to MTDā€“ TFAMĆ¾mtDNA 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 MTDā€“TFAM protein alone or MTDā€“TFAMĆ¾mtDNA complex increased expression of [[PGC-1Ī±]], 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.
|abstract=Neurodegeneration in Parkinsonā€™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 (ā€˜ā€˜ProtoFectionā€™ā€™) 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. MTDā€“TFAM (MTDĀ¼PTDĆ¾MLSĀ¼ā€˜ā€˜mitochondrial transduction domainā€™ā€™) binds mtDNA and rapidly transports it across plasma membranes to mitochondria. For therapeutic proof-of-principle we tested ProtoFection technology in Parkinsonā€™s disease cybrid cells, using mtDNA generated from commercially available human genomic DNA (gDNA; Roche). Nine to 11 weeks after single exposures to MTDā€“ TFAMĆ¾mtDNA 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 MTDā€“TFAM protein alone or MTDā€“TFAMĆ¾mtDNA complex increased expression of [[PGC-1Ī±]], 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.
|keywords=SH-SY5Y neuroblastoma-derived cells
|mipnetlab=US VA Richmond Bennett JP, US VA Richmond Iyer S
|mipnetlab=US VA Richmond Bennett JP, US VA Richmond Iyer S
|discipline=Biomedicine, Pharmacology; Biotechnology
|discipline=Biomedicine, Pharmacology; Biotechnology
}}
}}
{{Labeling
{{Labeling
|diseases=Parkinson's
|organism=Human
|organism=Human
|tissues=Nervous system
|tissues=Nervous system, Neuroblastoma
|preparations=Intact organism
|preparations=Intact cells
|injuries=Mitochondrial disease
|couplingstates=LEAK, ROUTINE, OXPHOS, ET
|diseases=Parkinson's
|pathways=ROX
|couplingstates=OXPHOS
|instruments=Oxygraph-2k
|instruments=Oxygraph-2k
|discipline=Biomedicine, Pharmacology; Biotechnology
|discipline=Biomedicine, Pharmacology; Biotechnology
}}
}}

Latest revision as of 13:52, 13 November 2017

Publications in the MiPMap
Keeney PM, Quigley CK, Dunham LD, Papageorge CM, Iyer S, Thomas RR, Schwarz KM, Trimmer PA, Khan SM, Portell FR, Bergquist KE, Bennett JP (2009) Mitochondrial gene therapy augments mitochondrial physiology in a Parkinson's disease cell model. Hum Gene Ther 20:897-907.

Ā» PMID: 19374590 Open Access

Keeney PM, Quigley CK, Dunham LD, Papageorge CM, Iyer S, Thomas RR, Schwarz KM, Trimmer PA, Khan SM, Portell FR, Bergquist KE, Bennett JP (2009) Hum Gene Ther

Abstract: Neurodegeneration in Parkinsonā€™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 (ā€˜ā€˜ProtoFectionā€™ā€™) 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. MTDā€“TFAM (MTDĀ¼PTDĆ¾MLSĀ¼ā€˜ā€˜mitochondrial transduction domainā€™ā€™) binds mtDNA and rapidly transports it across plasma membranes to mitochondria. For therapeutic proof-of-principle we tested ProtoFection technology in Parkinsonā€™s disease cybrid cells, using mtDNA generated from commercially available human genomic DNA (gDNA; Roche). Nine to 11 weeks after single exposures to MTDā€“ TFAMĆ¾mtDNA 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 MTDā€“TFAM protein alone or MTDā€“TFAMĆ¾mtDNA complex increased expression of PGC-1Ī±, 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. ā€¢ Keywords: SH-SY5Y neuroblastoma-derived cells

ā€¢ O2k-Network Lab: US VA Richmond Bennett JP, US VA Richmond Iyer S


Labels: Pathology: Parkinson's 

Organism: Human  Tissue;cell: Nervous system, Neuroblastoma  Preparation: Intact cells 


Coupling state: LEAK, ROUTINE, OXPHOS, ET  Pathway: ROX  HRR: Oxygraph-2k