Title:Mitochondrial Dysfunction Associated with mtDNA Mutation: Mitochondrial Genome Editing in Atherosclerosis Research
Volume: 33
Issue: 8
Author(s): Victoria A. Khotina*, Andrey Y. Vinokurov, Vasily V. Sinyov, Alexander D. Zhuravlev, Daniil Y. Popov, Vasily N. Sukhorukov, Igor A. Sobenin and Alexander N. Orekhov
Affiliation:
- Institute of General Pathology and Pathophysiology, 8 Baltiyskaya Street, Moscow, 125315, Russia
Keywords:
Atherosclerosis, mitochondrial DNA mutations, mitochondrial respiration, mitochondrial membrane potential, reactive oxygen species, lipid peroxidation.
Abstract:
Background: Atherosclerosis is a complex cardiovascular disease often associated
with mitochondrial dysfunction, which can lead to various cellular and metabolic
abnormalities. Within the mitochondrial genome, specific mutations have been implicated
in contributing to mitochondrial dysfunction. Atherosclerosis-associated
m.15059G>A mutation has been of particular interest due to its potential role in altering
mitochondrial function and cellular health.
Objective: This study aims to investigate the role of the atherosclerosis-associated
m.15059G>A mutation in the development of mitochondrial dysfunction in monocytelike
cells.
Methods: Monocyte-like cytoplasmic hybrid cell line TC-HSMAM1, which contains
the m.15059G>A mutation in mtDNA, was used. The MitoCas9 vector was utilized to
eliminate mtDNA copies carrying the m.15059G>A mutation from TC-HSMAM1 cybrids.
Mitochondrial membrane potential, generation of reactive oxygen species, and
lipid peroxidation levels were assessed using flow cytometry. Cellular reduced glutathione
levels were assessed using the confocal microscopy. The oxygen consumption
rate was measured using polarographic oxygen respirometry.
Results: The elimination of the m.15059G>A mutation resulted in a significant increase
in mitochondrial membrane potential and improved mitochondrial efficiency while also
causing a decrease in the generation of reactive oxygen species, lipid peroxidation, as
well as cellular bioenergetic parameters, such as proton leak and non-mitochondrial oxygen
consumption. At the same time, no changes were found in the intracellular antioxidant
system after the mitochondrial genome editing.
Conclusion: The presence of the m.15059G>A mutation contributes to mitochondrial
dysfunction by reducing mitochondrial membrane potential, increasing the generation of
reactive oxygen species and lipid peroxidation, and altering mitochondrial bioenergetics.
Elimination of the mtDNA containing atherogenic mutation leads to an improvement in
mitochondrial function.
