title: Mutagenesis of mitochondrial DNA in Fuchs endothelial corneal dystrophy.
abstract: Fuchs endothelial corneal dystrophy (FECD) is an age-related, slowly progressive disease, which may lead to loss of vision resulting from apoptosis of corneal endothelial (CE) cells, dysfunction of Descemet membrane (DM) and corneal edema. A growing body of evidence suggests that oxidative stress may play a major role in the pathogenesis of FECD and that mitochondria of CE cells are its main target. Mitochondrial DNA (mtDNA) is particularly prone to oxidative stress and changes in mtDNA were reported in FECD patients. In the present work we studied mtDNA damage and repair, mtDNA copy number, and the 4977bp common deletion in mtDNA in DM cells and peripheral blood lymphocytes (PBLs) isolated from FECD patients. PBLs from 35 FECD patients and 32 controls were challenged for 10min with hydrogen peroxide at 20μM and then left in a fresh medium for 3h, resulting in a decrease in mtDNA copy number in both groups. Damage to mtDNA was not fully repaired after 3h and the extent of remaining lesions was significantly higher in the patients than the controls. We observed a higher copy number and an increased extent of mtDNA damage as well as a higher ratio of the common 4977bp deletion in DM cells of FECD patients than the controls. Our results confirm that mutagenesis of mtDNA may be involved in FECD pathogenesis and disturbance in mtDNA sensitivity to damaging agent as well as changes in mtDNA damage repair along with alternations in mtDNA copy number may underline this involvement.
authors.affiliation: Department of Molecular Genetics, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland.
authors.forename: P
authors.initials: P
authors.lastname: Czarny
authors.affiliation: Department of Molecular Genetics, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Seda
authors.affiliation: Department of Ophthalmology, Medical University of Warsaw and Samodzielny Publiczny Kliniczny Szpital Okulistyczny, Sierakowskiego 13, 03-710 Warsaw, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Wielgorski
authors.affiliation: Department of Ophthalmology, Medical University of Warsaw and Samodzielny Publiczny Kliniczny Szpital Okulistyczny, Sierakowskiego 13, 03-710 Warsaw, Poland.
authors.forename: E
authors.initials: E
authors.lastname: Binczyk
authors.affiliation: Department of Molecular Genetics, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland.
authors.forename: B
authors.initials: B
authors.lastname: Markiewicz
authors.affiliation: Department of Molecular Genetics, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland.
authors.forename: E
authors.initials: E
authors.lastname: Kasprzak
authors.affiliation: University of Málaga, Malaga, Spain.
authors.forename: M P
authors.initials: MP
authors.lastname: Jiménez-García
authors.affiliation: Warsaw Eye Bank, Al. Solidarnosci 67, 03-401 Warsaw, Poland; Ophthalmology Clinic, Medical Center of Postgraduate Education, ul. Czerniakowska 231, 00-416 Warsaw, Poland.
authors.forename: I
authors.initials: I
authors.lastname: Grabska-Liberek
authors.affiliation: Department of Orthodontics, Medical University of Lodz, Pomorska 251, 92-216 Lodz, Poland.
authors.forename: E
authors.initials: E
authors.lastname: Pawlowska
authors.affiliation: Department of Molecular Genetics, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland.
authors.forename: J
authors.initials: J
authors.lastname: Blasiak
authors.affiliation: Department of Ophthalmology, Medical University of Warsaw and Samodzielny Publiczny Kliniczny Szpital Okulistyczny, Sierakowskiego 13, 03-710 Warsaw, Poland.
authors.forename: J
authors.initials: J
authors.lastname: Szaflik
authors.affiliation: Department of Ophthalmology, Medical University of Warsaw and Samodzielny Publiczny Kliniczny Szpital Okulistyczny, Sierakowskiego 13, 03-710 Warsaw, Poland. Electronic address: szaflik@ophthalmology.pl.
authors.forename: J P
authors.initials: JP
authors.lastname: Szaflik
mesh_terms: D000368:Aged; D017209:Apoptosis; D016022:Case-Control Studies; D056915:DNA Copy Number Variations; D004249:DNA Damage / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics*; D005260:Female; D005642:Fuchs' Endothelial Dystrophy / Q000235:genetics* / Q000473:pathology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D008928:Mitochondria / Q000235:genetics / Q000473:pathology*; D016296:Mutagenesis; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D017384:Sequence Deletion
chemical_list: D004272:DNA, Mitochondrial; D016877:Oxidants; D006861:Hydrogen Peroxide
keywords: 4977bp mtDNA common deletion; AMD; AS-OCT; CD; CE; DM; Descemet membrane; ETC; FECD; Fuchs endothelial corneal dystrophy; IVCM; Oxidative stress; PBLs; PBS; ROS; SLR rt-PCR; age-related macular degeneration; anterior segment optical coherence tomography; common deletion; corneal endothelial; electron transport chain; in vivo confocal microscopy; mitochondrial DNA; mtDNA; mtDNA copy number; mtDNA damage and repair; peripheral blood lymphocytes; phosphate buffered saline; qPCR; quantitative PCR; reactive oxygen species; semi-long rt-PCR; tRNA; transfer RNA