title: Reduction of infarct size by the therapeutic protein TAT-Ndi1 in vivo.
abstract: Lethal myocardial ischemia-reperfusion (I/R) injury has been attributed in part to mitochondrial respiratory dysfunction (including damage to complex I) and the resultant excessive production of reactive oxygen species. Recent evidence has shown that reduced nicotinamide adenine dinucleotide-quinone internal oxidoreductase (Ndi1; the single-subunit protein that in yeast serves the analogous function as complex I), transduced by addition of the TAT-conjugated protein to culture media and perfusion buffer, can preserve mitochondrial function and attenuate I/R injury in neonatal rat cardiomyocytes and Langendorff-perfused rat hearts. However, this novel metabolic strategy to salvage ischemic-reperfused myocardium has not been tested in vivo. In this study, TAT-conjugated Ndi1 and placebo-control protein were synthesized using a cell-free system. Mitochondrial uptake and functionality of TAT-Ndi1 were demonstrated in mitochondrial preparations from rat hearts after intraperitoneal administration of the protein. Rats were randomized to receive either TAT-Ndi1 or placebo protein, and 2 hours later all animals underwent 45-minute coronary artery occlusion followed by 2 hours of reperfusion. Infarct size was delineated by tetrazolium staining and normalized to the volume of at-risk myocardium, with all analysis conducted in a blinded manner. Risk region was comparable in the 2 cohorts. Preischemic administration of TAT-Ndi1 was profoundly cardioprotective. These results demonstrate that it is possible to target therapeutic proteins to the mitochondrial matrix and that yeast Ndi1 can substitute for complex I to ameliorate I/R injury in the heart. Moreover, these data suggest that cell-permeable delivery of mitochondrial proteins may provide a novel molecular strategy to treat mitochondrial dysfunction in patients.
authors.affiliation: 1Cardiovascular Research Institute, Wayne State University School of Medicine, Detroit, MI, USA.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Mentzer
authors.forename: Joseph
authors.initials: J
authors.lastname: Wider
authors.forename: Cynthia N
authors.initials: CN
authors.lastname: Perry
authors.forename: Roberta A
authors.initials: RA
authors.lastname: Gottlieb
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D042967:Electron Transport Complex I / Q000096:biosynthesis; D005260:Female; D015696:Gene Products, tat / Q000494:pharmacology*; D008929:Mitochondria, Heart / Q000378:metabolism; D009203:Myocardial Infarction / Q000503:physiopathology*; D015428:Myocardial Reperfusion Injury / Q000517:prevention & control*; D009243:NAD / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D015696:Gene Products, tat; D009243:NAD; D042967:Electron Transport Complex I
keywords: acute myocardial infarction; cardioprotection; complex I; heart disease; ischemia/reperfusion injury; mitochondria; myocardial protection