Zukhra B. Khasanova

Studies of the human aortic intima by a direct quantitative assay of mutant alleles in the mitochondrial genome

A mutant allele quantitative assay was developed to study somatic mitochondrial mutations associated with human diseases. This assay may be used in the clinical diagnostics for diseases associated with somatic mutations. To detect somatic mutations associated with atherosclerotic lesions of the aortal intima, we analyzed 40 mitochondrial mutations previously identified in several pathological conditions. 10 mutations associated with lipofibrosis plaques were found in mitochondrial genes that encode rRNA 12S, tRNA-Leu (UUR recognition codon), tRNA-Leu (CUN recognition codon), subunits of 1, 2, 5, and 6 NADH-dehydrogenase, and cytochrome B.

Mutations of mitochondrial genome in carotid atherosclerosis

With aim of detection the spectrum of mitochondrial DNA mutations in patients with carotid atherosclerosis from Moscow Region, we used a Roche 454 high-throughput sequencing of the whole mitochondrial genome. We have found that the presence of a number of homoplasmic mitochondrial DNA mutations in genes of 16S ribosomal RNA, subunits 2, 4, and 5 NADH dehydrogenase, subunits 1 and 2 cytochrome C oxidase, subunit 6 ATP-synthase, tRNA- Leu 2 and cytochrome B differed between conventionally healthy participants of the study and patients with carotid atherosclerosis. We also found heteroplasmic mutations, including insertions one or several nucleotides, that occurred more frequently in mitochondrial DNA of conventionally healthy participants of the study or patients with atherosclerotic lesions.

Role of Mitochondrial Genome Mutations in Pathogenesis of Carotid Atherosclerosis

Mutations of mtDNA, due to their higher frequency of occurrence compared to nuclear DNA mutations, are the most promising biomarkers for assessing predisposition of the occurrence and development of atherogenesis. 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. The sample size was 700 people. The sample members were diagnosed with “atherosclerosis” on the basis of ultrasonographic examination and biochemical and molecular cell tests. DNA was isolated from blood leukocyte samples of the study participants. PCR fragments of DNA, containing the region of 11 investigated mutations, were pyrosequenced. The heteroplasmy level of these mutations was detected. Statistical analysis of the obtained results was performed using the software package SPSS 22.0. According to the obtained results, an association of mutations m.652delG, m.3336C>T, m.12315G>A, m.14459G>A m.15059G>A with carotid atherosclerosis was found. These mutations can be biomarkers for assessing predisposition to this disease. Additionally, two single nucleotide substitutions (m.13513G>A and m.14846G>A), negatively correlating with atherosclerotic lesions, were detected. These mutations may be potential candidates for gene therapy of atherosclerosis and its risk factors.

1A.06: MITOCHONDRIAL DNA HAPLOGROUP H IS ASSOCIATED WITH SUBCLINICAL CAROTID ATHEROSCLEROSIS IN RUSSIAN POPULATION.

Objective: It is known that type of mitochondrial haplogroup, based on the combination of inherited mtDNA mutations, may influence the progression of various multifactorial diseases. For example, belonging to haplogroup H is associated with early myocardial infarction in the population of Asturias (northern Spain). Aim of this study was to identify the relationship between the type of mitochondrial haplogroup and presence of subclinical atherosclerosis and hypertension in Russian population. Design and method: A total of 80 persons from Moscow region (Russia) were included in the study. 45 study participants without CHD or myocardial infarction had ultrasonographically detected atherosclerotic lesions of the carotid arteries, others were controls without atherosclerosis. 32 patients had arterial hypertension. DNA was isolated from blood and the enrichment of mitochondrial DNA was performed. Detection of mtDNA haplogroups was made on the basis of Phylotree and MITOMAP databases and using Mitotool software on the data of mtDNA full sequences obtained by high-throughput sequencing of the mitochondrial genome using Roche 454 technology with GS Junior Titanium system. Statistical analysis was performed using IBM SPSS Statistics v.21.0 software. Results: Mitochondrial haplogroups H, U, T and J were the most common in the observed sample (85.7% of cases), what corresponds to the general Russian population data. It was found that belonging to haplogroup H is associated with an increased risk of atherosclerosis (x2 = 3.97, p = 0.046; OR = 2.76, 95 % CI 1.01–7.58). 2706A and 7028C variants, that are markers of mitochondrial haplogroup H, were more common in atherosclerotic patients (p < 0.05), which proofs the role of this haplogroup as a marker for suspectability to atherosclerotic-related diseases. There were no statistically significant evidence that any other haplogroups are associated with atherosclerosis and hypertension in the observed sample. Conclusions: Results of our study based on NGS data showed that haplogroup H is associated with carotid subclinical atherosclerosis and not associated with hypertension in Russian population.

Dataset of mitochondrial genome variants associated with asymptomatic atherosclerosis.

This dataset report is dedicated to mitochondrial genome variants associated with asymptomatic atherosclerosis. These data were obtained using the method of next generation pyrosequencing (NGPS). The whole mitochondrial genome of the sample of patients from the Moscow region was analyzed. In this article the dataset including anthropometric, biochemical and clinical parameters along with detected mtDNA variants in patients with carotid atherosclerosis and healthy individuals was presented. Among 58 of the most common homoplasmic mtDNA variants found in the observed sample, 7 variants occurred more often in patients with atherosclerosis and 16 variants occurred more often in healthy individuals.

Response to: Comment on “Role of Mitochondrial Genome Mutations in Pathogenesis of Carotid Atherosclerosis”

15a 3rd Cherepkovskaya Str, National Medical Research Center of Cardiology, Moscow 121552, Russia 8 Baltiyskaya Str, Institute of General Pathology and Pathophysiology, Moscow 125315, Russia K.I. Skryabin Moscow State Academy of Veterinary Medicine and Biotechnology-MVA, Moscow 109472, Russia Department of Genetics, 105/42 B. Sadovaya Str, Southern Federal University, Rostov-on-Don 344006, Russia Institute for Atherosclerosis Research, Skolkovo Innovative Centre, Moscow 121609, Russia

Data on association of mitochondrial heteroplasmy and cardiovascular risk factors: Comparison of samples from Russian and Mexican populations

Despite the fact that the role of mitochondrial genome mutations in a number of human diseases is widely studied, the effect of mitochondrial heteroplasmy in the development of cardiovascular disease has not been adequately investigated. In this study, we compared the heteroplasmy levels of mtDNA from leukocytes for m.3256C>T, m.3336T>C, m.12315G>A, m.5178C>A, m.13513G>A, m.14459G>A, m.14846G>A, m.15059G>A, m.652insG and m.1555A>G mutations in CVD-free subjects and CVD patients in samples derived from Russian and Mexican populations. It was demonstrated that heteroplasmy level of m.5178C>A was associated with CVD in Russian men, and m.14459G>A – in Russian women. Mitochondrial heteroplasmy level of m.13513G>A and m.652insG were associated with CVD in Mexican men, and only m.652insG– in Mexican women. The levels of heteroplasmy for mitochondrial mutations m.3336T>C, m.5178C>A, m.14459G>A, m.14846G>A and m.1555A>G were significantly higher in CVD-free Mexican men, and for m.3256C>T, m.3336T>C, and m.14459G>A – in CVD-free Mexican women.