Background and Aims: Tandem telomere repeats serve to maintain the stability of the nuclear genome. Telomere shortening is associated with factors such as unfavorable environmental factors and various diseases; i.e. with states characterized by the intensification of oxidative stress. The aim of this work was to study the association of telomere repeat length variants with coronary heart disease with old myocardial infarction (CHD with old MI). Methods: We analyzed relative telomeric repeat length (RTRL) variants in 150 DNA samples from samples of patients with CHD with old MI and 150 apparently healthy study participants by real-time PCR method on a BIO-RADCFX 96 Real-Time System amplifier. The results are presented as a percentage of the calibrator. DNA isolated from HeLa cell line was used as a calibrator. It should be noted that the RTRL variants have designations, depending on their relative length in percent to the calibrator. Results: According to the obtained results, the mean value of RTRL was significantly lower in patients with coronary heart disease with old myocardial infarction (by 25.0%), compared to apparently healthy study participants. Correlation analysis revealed an association of CHD with old MI with 5 RTRL variants: RTRL-46, RTRL-49, RTRL-51, RTRL-53 and RTRL-56. At the same time, 6 RTRL variants have a protective effect in CHD with old MI: RTRL-63, RTRL-65, RTRL-68, RTRL-70, RTRL-71 and RTRL-73. Conclusions: Five variants of RTRL are associated with CHD with old MI. This study was supported by Russian Science Foundation (Grant # 20-15-00364).

Background and Aims : The aim of the present study was to create cybrid lines with mtDNA mutation m.13513G>A. In our previous studies, it was found that, after reaching the threshold level of heteroplasmy, mutation m.13513G>A had a protective effect in atherosclerosis (33,5%).

Background and Aims : The aim of this study was a comparative electron microscopic analysis of mitochondria from lipofibrous plaques and normal aortic intima.Methods: Electron microscopic analysis of normal segments of intima and atherosclerotic plaques of aortas tissue samples was conducted using microscope Hitachi H7000. We investigated 315 mitochondria from cells of 38 aortas, 156 mitochondria of them were from atherosclerotic plaques and 159 mitochondria were from normal intima.Results: According to the obtained results, only 2% of mitochondria from normal intima (3 from 159) had changes in mitochondrial structure. At the same time, such changes were present in 15% of mitochondria from lipofibrous plaques of aortas (23 from 156). In mitochondria from atherosclerotic plaques, changes in the structure of cristae, edema of matrix, vacuole-like and myelin-like structures formation were observed.Conclusions: In mitochondria from atherosclerotic plaques, destructive changes of cristae, matrix edema, vacuole-like and myelin-like structures were found. The presence of structural changes in mitochondria of calls from atherosclerotic lesions allows us to suppose that there can be disorders on biochemical and genetic level in mitochondria. This study was supported by Russian Science Foundation (Grant # 20-15-00364). Background and Aims : The aim of this study was a comparative electron microscopic analysis of mitochondria from lipofibrous plaques and normal aortic intima. Methods: Electron microscopic analysis of normal segments of intima and atherosclerotic plaques of aortas tissue samples was conducted using microscope Hitachi H7000. We investigated 315 mitochondria from cells of 38 aortas, 156 mitochondria of them were from atherosclerotic plaques and 159 mitochondria were from normal intima. Results: According to the obtained results, only 2% of mitochondria from normal intima (3 from 159) had changes in mitochondrial structure. At the same time, such changes were present in 15% of mitochondria from lipofibrous plaques of aortas (23 from 156). In mitochondria from atherosclerotic plaques, changes in the structure of cristae, edema of matrix, vacuole-like and myelin-like structures formation were observed. Conclusions: In mitochondria from atherosclerotic plaques, destructive changes of cristae, matrix edema, vacuole-like and myelin-like structures were found. The presence of structural changes in mitochondria of calls from atherosclerotic lesions allows us to suppose that there can be disorders on biochemical and genetic level in mitochondria. This study was supported by Russian Science Foundation (Grant # 20-15-00364).

Background: The present review article considers some chronic diseases of vascular and metabolic genesis, the causes of which may be mitochondrial dysfunction. Very often, in the long course of the disease, complications may occur, leading to myocardial infarction or ischemic stroke and, as a result, death. In particular, a large percentage of human deaths nowadays belongs to cardiovascular diseases, such as coronary heart disease (CHD), arterial hypertension, cardiomyopathies, and type 2 diabetes mellitus. Objective: The aim of the present review was the analysis of literature sources, devoted to an investigation of a link of mitochondrial DNA mutations with chronic diseases of vascular and metabolic genesis. Results: The analysis of literature indicates the association of the mitochondrial genome mutations with coronary heart disease, type 2 diabetes mellitus, hypertension, and various types of cardiomyopathies. Conclusion: The detected mutations can be used to analyze the predisposition to chronic diseases of vascular and metabolic genesis. They can also be used to create molecular-cell models necessary to evaluate the effectiveness of drugs developed for the treatment of these pathologies. MtDNA mutations associated with the absence of diseases of vascular and metabolic genesis could be potential candidates for gene therapy of the said diseases.

Background and Aims: Cytoplasmatic hybrids (cybrids) are widely used in scientific researches as a cell model for investigating different human diseases. The aim of the present study was creation of cybrid lines with mtDNA mutation m.14846G>A. In our preliminary studies, it was found that, after reaching the threshold level of heteroplasmy, mutation m.14846G>A had a protective effect in atherosclerosis (17.5%).

Background and Aims: The aim of the study was to identify heteroplasmic mutations mtDNA associated with atherosclerotic plaques in carotid arteries (AP CA) by the NGS (next-generation sequencing) method.

Введение. В настоящее время все больший интерес ученых мира вызывают цибридные клеточные модели, которые являются одним из лучших объектов для изучения патологических процессов в организме человека. Например, сотрудниками нашей лаборатории были впервые созданы подобные модели для изучения протективного эффекта некоторых мутаций митохондриального генома, защищающих организм человека от дисфункции митохондрий и атеросклеротических поражений. Цель: исследования - создание цибридных культур с высоким уровнем гетероплазмии по мутации митохондриального генома m.1555A>G, локализованной в кодирующем регионе митохондриального генома человека в гене MT-RNR1. В наших предварительных исследованиях было установлено, что пороговый уровень гетероплазмии мутации m.1555A>G имеет при атеросклерозе протективный эффект. Методика. Цибридные культуры были созданы путем слияния rho0(безмитохондриальных)-клеток и митохондрий из тромбоцитов с высоким уровнем гетероплазмии исследуемых мутаций. Для получения безмитохондриальных клеток была использована культура моноцитарного происхождения THP-1. Результаты. Получены 4 цибридные клеточные линии, содержащие мутацию m.1555A>G с уровнем гетероплазмии выше порогового значения. Заключение. В данной работе были созданы 4 цибридные культуры с высоким уровнем гетероплазмии по мутации мтДНК m.1555A>G, имеющей при атеросклерозе протективный эффект. Полученные цибридные клеточные линии могут служить моделями для отработки методов генотерапии у пациентов с атеросклерозом. Кроме того, с помощью данных цибридных клеточных моделей можно будет изучать молекулярно-клеточные механизмы, защищающие клетки от митохондриальной дисфункции. Introduction. Cybrid cell models are one of the best objects for studying pathological processes in the human body, and they are of increasing interest to scientists worldwide. Our laboratory was the first to create such models for studying the protective effect of mutations in the mitochondrial genome that protect the human body from mitochondrial dysfunction and atherosclerotic lesions. Aim: To create cybrid cultures with a high heteroplasmy level for the mitochondrial genome mutation m.1555A>G localized within the coding region of the human mitochondrial genome in the MT-RNR1 gene. Preliminary studies showed that the threshold heteroplasmy level for the m.1555A>G mutation has a protective effect in atherosclerosis. Methods. Cybrid cultures were created by fusion of rho0 (mtDNA-depleted) cells and mitochondria from platelets with a high heteroplasmy level for the studied mutations. To obtain mtDNA-free cells, a culture of monocytic origin, THP-1, was used. Results. We obtained four cybrid cell lines containing the m.1555A>G mutation with a heteroplasmy level above the threshold value. Conclusion. Four cybrid cultures with a high heteroplasmy level for the mtDNA mutation m.1555A>G were created. These cybrid cell lines can serve as models for developing methods of gene therapy for patients with atherosclerosis. In addition, using these cybrid cell models, it will be possible to study molecular and cellular mechanisms that protect cells from mitochondrial dysfunction.

Введение. Геронтология - это наука о причинах биологического старения организма. Ее составной частью является гериатрия - наука о болезнях и факторах, вызывающих старение. Полагаем, что митохондриальные мутации, затрагивающие гены белковых субъединиц дыхательной цепи, а также транспортных и рибосомальных РНК, могут являться одним из факторов старения индивидов. При этом, согласно данным литературы, проблема связи мутаций митохондриального генома со старением индивидов изучена недостаточно. Цель исследования - анализ ассоциации уровня гетероплазмии мутаций мтДНК с возрастом индивидов. Методика. Объектом исследования была выборка жителей Московского региона состоящая из 712 участников. ДНК выделяли из образцов лейкоцитов крови участников исследования с помощью метода фенол-хлороформной экстракции. Была проведена ПЦР фрагментов ДНК, содержащих область исследованных мутаций. Полученные амплификаты были пиросеквенированы. Анализ уровня гетероплазмии данных мутаций был проведен с помощью количественного метода, разработанного авторами статьи. Для подсчета уровня гетероплазмии мутаций по данным пирограммы использовалась ранее разработанная авторами формула. Результаты. Показано, что с возрастом индивидов положительно коррелируют однонуклеотидные замены митохондриального генома m.12315G>A, m.14459G>A и m.15059G>A (p≤0,05) и отрицательно - m.1555A>G и m.14846G>A (p≤0,05). Мутации m.652insG и m.13513G>A отрицательно коррелируют с возрастом на уровне значимости p≤0,1. Заключение. Обнаружены 3 однонуклеотидные замены m.14459G>A, m.15059G>A и m.12315G>A генов MT-ND6, MT-CYTB и MT-TL2 (соответственно) ассоциированные со старением организма человека. Данные мутации могут быть использованы для создания молекулярно-клеточных моделей старения. В то же время, выявлены мутации m.1555A>G, m.652insG (ген MT-RNR1), m.14846G>A (ген CYTB) и m.13513G>A (ген MT-ND5), отрицательно коррелирующие с возрастом индивидов. Данные мутации могут быть использованы при генотерапии для замедления старения людей. Introduction. Gerontology is a branch of science that focuses on the causes of biological aging. Its clinical component is geriatrics, a branch of medical science that focuses on diseases and factors causing aging. We hypothesize that mitochondrial mutations that affect genes of protein subunits of the respiratory chain, and, also, transport and ribosomal RNAs may be factors of aging. Meanwhile, according to the literature, the connection of mitochondrial genome mutations with aging has not been adequately studied. Aim. To analyze the association of heteroplasmy level in mtDNA mutations with the age of individuals. Methods. A sample of inhabitants of the Moscow region, composed of 712 study participants, was enrolled in the study. DNA was isolated from blood leukocytes by phenol chloroform extraction, and PCR of DNA fragments, containing the region of the studied mutations was performed. The obtained amplificates were pyrosequenced. These mutations were analyzed for the heteroplasmy level using a quantitative method developed by the authors. To determine the heteroplasmy level of mutations from the pyrogram data, a formula was used that had earlier been developed by the authors. The results were statistically analyzed with the software package, IBM SPSS Statistics, version 27.0 (SPSS Inc., USA). Results. Analysis of mutations showed that single nucleotide substitutions of the mitochondrial genomes m.12315G>A, m.14459G>A and m.15059G>A correlated positively with subject age (p≤0.05) whereas m.1555A>G and m.14846G>A correlated negatively (p≤0.05). Mutations m.652insG and m.13513G>A tended to correlate negatively with aging (p≤0.1). Conclusion. Three single nucleotide substitutions were found: m.14459G>A, m.15059G>A, and m.12315G>A of genes MT-ND6, MT-CYTB, and MT-TL2, respectively, that are associated with human aging. Thus, these mutations may be used for creating molecular cellular models of aging. At the same time, mutations m.1555A>G, m.652insG (gene MT-RNR1), m.14846G>A (gene CYTB), and m.13513G>A (gene MT-ND5) were detected, which correlate with aging negatively. These mutations may be used in gene therapy to slow aging.

Background and Aims: The aim of the present study was a creation of cybrid cultures containing mitochondrial genome mutation m.5178C>A, associated with atherosclerosis. This mutation was localized in gene MT-ND2.

Введение. Цибридные клеточные модели наиболее перспективны для изучения патогенеза различных заболеваний. Авторами статьи впервые были созданы такие модели для изучения митохондриальной дисфункции и патологических процессов, развивающихся при атеросклерозе. Цель работы - создание цибридных культур с высоким уровнем гетероплазмии по мутации митохондриального генома m.12315G>A. В предварительных исследованиях авторами статьи было установлено, что пороговый уровень гетероплазмии мутации m.12315G>A ассоциирован с атеросклерозом. Методика. Цибридные культуры создавали путем слияния безмитохондриальных клеток (rho0) и митохондрий из тромбоцитов участников исследования с высоким уровнем гетероплазмии исследуемых мутаций. Для создания rho0-клеток была взята культура моноцитарного происхождения THP-1. Безмитохондриальные клетки были получены с помощью метода M. Kинга и Г. Аттарди. Тромбоциты выделяли из цельной крови участников исследования. Для этого был применен метод центрифугирования в градиенте плотности фиколла-урографина. Для получения цибридных культур клеток была использована методика «ПЭГ-слияния». В созданных безмитохондриальных и цибридных клеточных культурах был проведен количественный анализ копий митохондриального генома. Согласно результатам данного анализа было подтверждено либо отсутствие митохондрий (rho0-клетки), либо их наличие (цибриды). Количество копий мтДНК детектировалось с помощью реал-тайм ПЦР в присутствии красителя SYBR Green I. Результаты. Получены 4 цибридные клеточные линии, содержащие мутацию m.12315G>A с уровнем гетероплазмии выше порогового значения. Заключение. Созданы 4 цибридные культуры с высоким уровнем гетероплазмии по мутации митохондриального генома m.12315G>A. Полученные цибридные клеточные линии могут служить моделями для изучения молекулярно-клеточных механизмов митохондриальной дисфункции при атеросклерозе и других сердечно-сосудистых заболеваниях. Цибридные культуры можно использовать для моделирования атерогенеза, а также для подбора патогенетически обоснованной лекарственной терапии при атеросклерозе. Introduction. Cybrid cell models are most promising for studying pathological mechanisms in different diseases. The authors for the first time created such models for studying mitochondrial dysfunction and pathological processes underlying atherosclerosis. Aim. Creation of cybrid cultures with a high heteroplasmy level for mitochondrial genome mutation m.12315G>A. A preliminary study by the authors showed that the heteroplasmy level of mutation m.12315G>A was associated with atherosclerosis. Methods. Cybrid cultures were created by fusing non-mitochondrial cells (rho0) and mitochondria from platelets of study participants with a high heteroplasmy level of the mutations under study. A THP-1 culture of monocytic origin was used to create rho0 cells. Non-mitochondrial cells were obtained using the M. King and G. Attardi method. Platelets were extracted from whole blood of study participants with Ficoll-Urografin density gradient centrifugation. Cybrid cell cultures were obtained by the PEG-mediated fusion method. In the created non-mitochondrial and cybrid cell cultures, quantitative analysis of mitochondrial genome copies was performed. This analysis confirmed either the absence of mitochondria (rho0-cells) or their presence (cybrids). The mtDNA copies were quantified using real-time PCR in the presence of the SYBR Green I stain. Results. Four cybrid cell lines were obtained, which contained the m.12315G>A mutation with heteroplasmy levels higher than the threshold level. Conclusion. Four cybrid cultures were created with a high heteroplasmy level for the mitochondrial genome mutation m.12315G>A. The obtained cell lines can be used as models for studying molecular cellular mechanisms of mitochondrial dysfunction in atherosclerosis and cardiovascular diseases. In addition, they may be useful for modeling atherogenesis in cells and for selecting therapy for patients with atherosclerosis.

Background and Aims: The aim of the present study was to investigate the speed of oxygen consumption in two cybrid lines with different mutational burden of mtDNA.

Background and Aims: Atherosclerosis of human major vessels is often a morphological basis of mortality from cardiovascular diseases. In this pathology intima of arteries is damaged, luminal occlusion occurs and blood supply to organs is deteriorated. Atherosclerosis is difficult to recognize in early stages. Molecular genetic markers could help the diagnostics of this disease. The aim of the present article was the detection of threshold heteroplasmy level of mtDNA mutations, after reaching which in a patient appeared atherosclerotic lesions. Besides, this parameter was detected for mutations, in which after reaching threshold heteroplasmy level, a protective antiatherogenic effect started to appear.

Background and Aims: The aim of the present article was an analysis of correlation of several mitochondrial genome mutations with carotid atherosclerosis. Leukocytes from blood of study participants from Moscow polyclinics were used as research material.

Background and Aims: To study the characteristics of the distribution and variability of mitochondrial genome mutations in various human tissues.

Background and Aims: In this study mtDNA mutations in atherosclerotic plaques of the carotid arteries (AP CA) in patients from Novosibirsk were analysed. The aim of the study was to compare samples from Novosibirsk and Moscow by mutation spectrum.

Background and Aims: One of the earliest manifestations of atherosclerotic lesions is the accumulation of lipid in arterial intimal cells. The major source of lipids is low density lipoprotein (LDL), which has undergone proatherogenic modification, possibly by desialylation. It is necessary to study the characteristics and the nature of neuraminidase (sialidase) activity in human blood plasma.

Aim: In this pilot study, the dependence of oxygen consumption rate on the heteroplasmy level of mitochondrial genome mutations associated with atherosclerosis was analyzed.

Aim: The aim of the study was to identify homoplasmic mutations mtDNA associated with atherosclerotic lesions by the NGS (next-generation sequencing) method.

Autophagy is a highly conservative process of degeneration during which intracellular components including soluble macromolecules (e. g., nucleic acids, proteins, carbohydrates and lipids) and dysfunctional organelles (e. g., mitochondria, peroxisomes, and the endoplasmatic reticulum) are degraded by a lysosome. Autophagy serves as a dynamic system of recycling proving cells with energy and building components. Because of it in cells of an organism new proteins and membranes can form contributing to survival of the individual under starvation conditions. Autophagy plays an important role in the genesis and development of multifactorial pathogenesis including atherosclerosis and its risk factors. The present article examines both a pathogenic and protective role of autophagy in such pathological processes. The article can be useful to molecular biologists and biochemists, as well as to professionals involved in the problems of atherosclerosis and cardiovascular diseases.

Aim. This review article describes literature sources devoted to the investigation of mitochondrial dysfunction using cytoplasmic hybrids (cybrids). The presented studies were carried out on cultures of cybrid cell lines HL60, MOL T-4, A549, 143B, HeLa, Arpe-19, HEK-293, SH-SY5Y and NT2. According to the analysis of scientific world literature, some of the most promising models for studying mitochondrial dysfunction are cell cultures without mitochondria (rho0) and cytoplasmic hybrids containing one or several mutations of mitochondrial genome. In the review scientific researches on studying biochemical and molecular cellular pathological processes in cybrid cells in various human diseases such as Alzheimer's disease and mild cognitive impairment, MERRF and MELAS syndromes, Leber's optic atrophy and Parkinson's disease were considered. Material dedicated to cybrids as potential models for the study of treatment possibilities was presented separately. Conclusion. The analyzed in the review rho0-cell cultures and cybrid lines containing mtDNA mutations may be models for the study of mitochondrial genome dysfunctions, biochemical and molecular cellular pathological processes. It is worth noting that in various cell cultures, similar tendencies are observed in functional activity changes of rho0-cell and cybrids compared with native cell lines. For example, such tendencies as reduction of oxygen consumption level, morphological changes of mitochondrial structure, resistance to apoptosis, reduction of ATP consumption level, increase in glucose consumption, activity deterioration of some respiratory chain complexes.