Targeted transgenic overexpression of mitochondrial thymidine kinase (TK2) alters mitochondrial DNA (mtDNA) and mitochondrial polypeptide abundance: transgenic TK2, mtDNA, and antiretrovirals.
Mitochondrial toxicity limits nucleoside reverse transcriptase inhibitors (NRTIs) for acquired immune deficiency syndrome. NRTI triphosphates, the active moieties, inhibit human immunodeficiency virus reverse transcriptase and eukaryotic mitochondrial DNA polymerase pol-gamma. NRTI phosphorylation seems to correlate with mitochondrial toxicity, but experimental evidence is lacking. Transgenic mice (TGs) with cardiac overexpression of thymidine kinase isoforms (mitochondrial TK2 and cytoplasmic TK1) were used to study NRTI mitochondrial toxicity. Echocardiography and nuclear magnetic resonance imaging defined cardiac performance and structure. TK gene copy and enzyme activity, mitochondrial (mt) DNA and polypeptide abundance, succinate dehydrogenase and cytochrome oxidase histochemistry, and electron microscopy correlated with transgenesis, mitochondrial structure, and biogenesis. Antiretroviral combinations simulated therapy. Untreated hTK1 or TK2 TGs exhibited normal left ventricle mass. In TK2 TGs, cardiac TK2 gene copy doubled, activity increased 300-fold, and mtDNA abundance doubled. Abundance of the 17-kd subunit of complex I, succinate dehydrogenase histochemical activity, and cristae density increased. NRTIs increased left ventricle mass 20% in TK2 TGs. TK activity increased 3 logs in hTK1 TGs, but no cardiac phenotype resulted. NRTIs abrogated functional effects of transgenically increased TK2 activity but had no effect on TK2 mtDNA abundance. Thus, NRTI mitochondrial phosphorylation by TK2 is integral to clinical NRTI mitochondrial toxicity.
Hosseini
Seyed H
SH
Department of Pathology, Emory University School of Medicine, 7117 Woodruff Memorial Building, 101 Woodruff Circle, Atlanta, GA 30322, USA.
Kohler
James J
JJ
Haase
Chad P
CP
Tioleco
Nina
N
Stuart
Tami
T
Keebaugh
Erin
E
Ludaway
Tomika
T
Russ
Rodney
R
Green
Elgin
E
Long
Robert
R
Wang
Liya
L
Eriksson
Staffan
S
Lewis
William
W
D000818:Animals; D004272:DNA, Mitochondrial / Q000187:drug effects / Q000378:metabolism; D004452:Echocardiography; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000187:drug effects / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008822:Mice, Transgenic; D046529:Microscopy, Electron, Transmission; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology; D010455:Peptides / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D018894:Reverse Transcriptase Inhibitors / Q000633:toxicity*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013385:Succinate Dehydrogenase / Q000187:drug effects; D013937:Thymidine Kinase / Q000187:drug effects* / Q000378:metabolism*; D019076:Transgenes; D015854:Up-Regulation
D004272:DNA, Mitochondrial; D007527:Isoenzymes; D010455:Peptides; D018894:Reverse Transcriptase Inhibitors; D013385:Succinate Dehydrogenase; C105339:thymidine kinase 2; D013937:Thymidine Kinase