Creation of Cybrid Cultures Containing mtDNA Mutations m.12315G>A and m.1555G>A, Associated with Atherosclerosis

Abstract

In the present work, a pilot creation of four cybrid cultures with high heteroplasmy level was performed using mitochondrial genome mutations m.12315G>A and m.1555G>A. According to data of our preliminary studies, the threshold heteroplasmy level of mutation m.12315G>A is associated with atherosclerosis. At the same time, for a mutation m.1555G>A, such a heteroplasmy level is associated with the absence of atherosclerosis. Cybrid cultures were created by fusion of rho0-cells and mitochondria from platelets with a high heteroplasmy level of the investigated mutations. To create rho0-cells, THP-1 culture of monocytic origin was taken. According to the results of the study, two cybrid cell lines containing mutation m.12315G>A with the heteroplasmy level above the threshold value (25% and 44%, respectively) were obtained. In addition, two cybrid cell lines containing mutation m.1555G>A with a high heteroplasmy level (24%) were obtained. Cybrid cultures with mtDNA mutation m.12315G>A can be used to model both the occurrence and development of atherosclerosis in cells and the titration of drug therapy for patients with atherosclerosis. With the help of cybrid cultures containing single nucleotide replacement of mitochondrial genome m.1555G>A, it is possible to develop approaches to the gene therapy of atherosclerosis.

The Role of Mitochondrial Dysfunction in Vascular Disease, Tumorigenesis, and Diabetes

“…Such models allow studying the effects of mtDNA mutations in a constant nuclear genetic background (Wilkins et al, 2014). The cybrid models of Parkinson's disease, MELAS, LHON, Alzheimer's disease, Leigh syndrome and atherosclerosis have been already developed and characterized (Arduíno et al, 2015;Jeong et al, 2015;Jiang et al, 2016;Sazonova et al, 2019). The importance of cybrid models is highlighted by the great phenotypic heterogeneity associated with mtDNA mutations described in these pathologies.…”

Some Molecular and Cellular Stress Mechanisms Associated with Neurodegenerative Diseases and Atherosclerosis

“…The authors of the article created two cybrid cultures, one of which contained an atherogenic mutation of the mitochondrial genome (m.12315G>A), and the other contained an antiatherogenic mutation mtDNA (m.1555G>A) [ 240 , 241 , 242 , 243 , 244 ]. These cultures was created on the basis of a monocytic origin culture of THP-1.…”

“…At the same time, in cybrid cells containing an antiatherogenic mutation, the level of oxygen consumption was higher than in the native THP-1 culture. We assume that this data may indicate a negative effect of oxidative stress on a cybrid culture containing an atherogenic mutation of the mitochondrial genome [ 240 , 241 , 242 , 243 , 244 ].…”

“…In addition, our research team found that the number of copies of mtDNA in a cybrid culture containing atherogenic mutation m.12315G>A was significantly lower than in a native culture of THP-1. We assume that this fact indicates the occurrence of replicative stress in this cybrid culture [ 240 , 241 , 242 , 243 , 244 ].…”

A Brief History of Mitochondrial Pathologies

“…Some mtDNA mutations from the most updated list of mutations potentially involved in atherosclerosis’ pathogenetic mechanisms [53] are presently under experimental test to verify their effective role in the disease’s onset. In fact, the creation of cybrids carrying different heteroplasmic levels of two mtDNA mutations, one associated with the presence and the other with the absence of atherosclerosis [54], will allow to thoroughly investigate the occurrence and the development of atherosclerosis in a quite simple cell model. This is the ultimate chronological step, but certainly not the last one along the pathway leading to the progressive understanding of the relevance of mtDNA mutations for monogenic as well as polygenic diseases.…”