To assess the contribution to survival of Alu insertions in the ACE, PLAT, COL13A1, LAMA2, CDH4, SEMA6A, PKHD1L1, STK38L, HECW1, and TEAD1 genes, which are candidates of aging and longevity, amid the senile physiological and pathological phenotype, an analysis of association with life expectancy was carried out. Survival and mortality data were obtained for 1382 elderly people who were selected from the sample of Tatars residing in the Republic of Bashkortostan (total 1790 people from 18 to 109 years). Mortality risk was higher among carriers of the STK38L Alu-insertion genotype (Ya5ac2145*II, HR = 2.07, P = 0.022). Alu insertion in the HECW1 and TEAD1 genes has demonstrated a survival protection effect (Ya5NBC182*II, HR = 0.71, P = 0.038 and Ya5ac2013*II, HR = 0.74, P = 0.035 respectively). The survival amid the persons with various clinical phenotypes was associated with the Alu polymorphism of the SEMA6A (Yb8NBC597*ID, HR = 0.54, P = 0.016 for the cerebrovascular diseases), TEAD1 (Ya5ac2013*II, HR = 0.57, P = 0.016 for the cardiovascular pathologies), and LAMA2 (Ya5-MLS19*ID, HR = 0.36, P = 0.03 for multimorbidity status) genes. Thus, the genes involved in the regulation of autophagy and apoptosis were associated with survival and longevity.

Myocardial infarction (MI) is a multifactorial polygenic disease that develops as a result of a complex interaction of numerous genetic factors and the external environment. Accordingly, the contribution of each of them separately is usually not large and may significantly depend on the state of other accompanying factors. The purpose of the study was to search for informative predictors of MI risk based on polygenic analysis of polymorphic variants of (1) the antioxidant defense enzyme genes PON1 (rs662), PON2 (rs7493), CAT (rs1001179), MSRA (rs10098474) and GSTP1 (rs1695); (2) the apoptosis genes CASP8 (rs3834129), TP53 (rs1042522) and BCL2 (rs12454712); and (3) the inflammation genes CRP (rs1205), CX3CR1 (rs3732378), IL6 (rs1800795) and CCL2 (rs1024611). 591 DNA samples were used in the study (280 patients with the onset at 30 to 60 years, with an average age of 46.02 ± 6.17, and 311 control subjects aged 30 to 62, with an average age of 44.65 ± 7.07). All the participants were male and Tatars by ethnicity. The logistic regression analysis with various models demonstrated associations with MI of polymorphic variants of the genes CX3CR1 (rs3732378) (overdominant model – G/G + A/A vs A/G P = 0.0002, OR = 1.9), MSRA (rs10098474) (dominant model – T/T vs T/C + C/C P = 0.015, OR = 1.51), CCL2 (rs1024611) (recessive model – P = 0.0007 – A/A + A/G vs G/G OR = 2.63), BCL2 (rs12454712) (log-additive model – *C allele, P = 0.005, OR = 1.38). Using the Monte Carlo method and Markov chains (APSampler), combinations of alleles/ genotypes of the studied polymorphic loci associated with a high risk of MI were obtained, which, in addition to those identified during single-locus analysis, contained polymorphic variants of the genes CASP8, TP53, CAT, PON2, CRP, IL6, GSTP1. Among the combinations obtained, a pairwise analysis of possible non-linear interactions between the identified combinations of alleles/genotypes was carried out, which showed synergistic interactions of the polymorphic variants CX3CR1*A/G and CASP8*I/I, MSRA*C and CRP*C, CAT*C/T and MSRA*C, CAT*C/T and CX3CR1*A contributing to the development of MI. Based on the results obtained using multivariate logistic regression analysis, a predictive model was built to assess the risk of developing MI, the predictive ability of which reached the value AUC = 0.71 (AUC – area under the curve in ROC analysis).

OBJECTIVE:Identification of a complex of genetic predictors of multiple sclerosis (MS) based on previously obtained results in genome-wide association studies of disease markers (GWAS markers) in a population of MS patients and healthy individuals of the Republic of Bashkortostan (Russian Federation) using polygenic detection.MATERIAL AND METHODS:The total study group consisted of 2048 people (641 patients with MS and 1407 healthy individuals) who permanently resided in the Republic of Bashkortostan and belonged to the Bashkir (n=325), Russian (n=772) or Tatar (n=951) nationalities. The analysis of association between MS and polymorphisms previously associated with the disease according to GWAS data was performed. Of the 641 MS patients, 247 were the subject of a 20-year prospective clinical follow-up.RESULTS:The C6orf10 rs3129934*T allele was most significantly associated with MS in Russians (OR=2.00, P=5.85·10-5) and Tatars (OR=2.38, P=8.61·10-7). An increased MS risk in Russians was also associated with the EOMES rs11129295*T (OR=1.56, P=0.007) and IL7R rs1494558*I (OR=1.61, P=0.003) alleles. Meta-analysis confirmed the association of the C6orf10 rs3129934*T, EOMES rs11129295*T and IL7R rs1494558*I alleles with MS in the total group, as well as revealed associations of the INAVA rs7522462*G, IL7R rs10624573*I, CD6 rs17824933*G, GPC5 rs9523762*A and GPR65 rs2119704*C alleles with the disease. Using polygenic analysis, we identified a complex predictor C6orf10 rs3129934*C + INAVA rs7522462*G + CD6 rs17824933*C with a pronounced protective effect against MS in the total group (OR=0.34, PFDR=2.65·10-7).CONCLUSION:We reproduced the association of eight polymorphisms (C6orf10 rs3129934, INAVA rs7522462, IL7R rs10624573, EOMES rs11129295, GPR65 rs2119704, GPC5 rs9523762, CD6 rs17824933 and CD58 rs2300747) with MS, previously identified in GWAS in European populations. Whole exome or genome sequencing may help to reveal the mechanisms underlying the pathogenesis of MS in populations of the Russian Federation.

With the purpose to search for predictors of aging and longevity we studied the pattern of the SOD1 and SOD2 genes polymorphic loci interaction in a sample of residents of the Republic of Bashkortostan, including 1592 individuals in age between 18 and 109 years old. We discovered the reduction of the chances to become the long-liver among carriers of the combination ofSOD2*rs4880*TT genotype and SOD1*rs2070424*G allele (OR=0.33, POR=0.02, SF=0.27, PSF=0.046 for elderly people and OR=0.31, POR=0.01, SF=0.28, PSF=0.041 for old seniors), as well as of the SOD2*rs4880*TT and SOD1*rs2070424*GA genotypes combination (OR=0.32, POR=0.01, SF=0.29, PSF=0.043 for the elderly people). The found alleles/genotypes combinations, which associated with the low activity of Cu/Zn-SOD and Mn-SOD, have a synergistic pattern of interaction and are unfavorable for the attainment the longevity.

Long non-coding RNA locus PVT1 produces multiple regulatory RNA molecules via alternative splicing, and therefore is a promising object for the development of targeted therapies of different disorders. Performing associationanalysis of genetic variants rs4410871 andrs759648 at the PVT1 locus with multiple sclerosis (MS) in the ethnic group of Russians, Tatars, and Bashkirs from the Volga-Ural region of Russia, we established an association between rs759648*С and MS in the group of Tatars (OR=1.42, PBonf0.046), confirmed by the results of meta-analysisin the three ethnic groups (OR=1.28, P = 0.015). Haplotypic analysis has revealed an association of the rs759648*С/rs4410871*С haplotype with MS in the total study group (χ2=6.083, Рperm =0.012), whilers4410871*C/C + rs759648* C/C combination conferred an increased risk of MS (OR = 2.71, Pperm = 0.027) in this groupaccording to the data yielded by the multilocus analysis. The results of our analysis indicate the significance of the rs759648 polymorphism located near the DNA sequence producing miR-1208. Further studies will help to elucidate the molecular mechanisms of the involvement of PVT1 gene in the etiopathogenesis of MS.

Цель. Цель исследования состояла в поиске информативных предикторов эффективности лечения РС для использования в персонализированной терапии заболевания на основании проспективного исследования РС в Республике Башкортостан.

Objective. To investigate the molecular mechanism underlying genetic susceptibility to essential hypertension (EH) using polygenic analysis of renin-angiotensin-aldosterone system (RAAS).Design and methods.Genotyping of renin (REN, rs2368564), angiotensinogen (AGT, rs4762), angiotensin II receptor type 1 (AGTR1, rs5186), chymase 1 (CMA1, rs1800875) and angiotensin-converting enzyme (ACE, rs1799752) polymorphic variants was performed in 346 patients with EH and 377 controls, Russians and Tatars by ethnic origin.Results. ACE rs1799752polymorphism was significantly associated with EH risk in Tatars (PBonf= 0,003) and in the total study group (PBonf= 4,09 x 10–5). Polygenic approach identified 12 genotypes and/or alleles combinations of RAAS genes polymorphisms, significantly associated with EH in the Tatars, and 6 patterns associated with EH in the total study group. The highest risk of disease in Tatar men was associated with REN rs2368564*T + AGTR1 rs5186*C/A + ACE rs1799752*D combination (OR = 16,64, PBonf= 0,001), in the total group — with REN rs2368564*T/C + CMA1 rs1800875*G combination (OR = 2,37, PBonf= 0,045).Conclusions. Our findings indicate that EH risk in men of Russian and Tatar ethnicity is significantly associated with ACE rs1799752 polymorphism, and the results of polygenic analysis demonstrate an association of the disease risk with genotype/allele combinations of polymorphic variants in REN (rs2368564), AGTR1 (rs5186), ACE (rs1799752), and CMA1 (rs1800875) genes.

The renin-angiotensin-aldosterone system plays an important role in the processes responsible for the formation of myocardial infarction (MI). It is involved in the regulation of oxidative stress, in the development of endothelial dysfunction and inflammatory response, in destabilization of atherosclerotic plaque, regulation of vascular tone and circulating blood volume. A significant part of these effects are mediated through the molecules of the JAK/STAT signaling pathway and the nuclear factor NF-kappa-B1. The aim of this study was to analyze associations of MI with combinations of alleles/genotypes by polymorphic markers rs310216*JAK1, rs12693591*STAT1, rs3212780*JAK3, rs2293152*STAT3, rs28362491*NFKB1, rs4762*AGT, rs2368564*REN, rs4343*ACE, rs5186*AGTR1, rs1800875*CMA1, rs4491175*LINC02748, rs1799998*CYP11B2, rs2285666*ACE2, rs1403543*AGTR2. The material for the study was the DNA of patients with onset MI at the age of 45 years (144) and the control group (152). All study participants were male, ethnically Russian. Combinations were searched using the APSampler software. The selection criteria for combinations were: a P value <0.05 after adjusting for multiple comparisons (Benjamini-Hochberg FDR procedure) and an OR value of more than 3 for high-risk markers and less than 0.3 for protective markers. As a result of the analysis, combinations of alleles with both increased and reduced risk of MI were obtained, in which allelic variants rs5186*AGTR1, rs2368564*REN, rs4491175*LINC02748, rs1800875*CMA1rs2285666*ACE2, rs1403543*AGTR2, rs12693591*STAT1, rs3212780*JAK3, rs2293152*STAT3 occur.

Background: Genome-wide association studies identified numerous susceptibility loci for multiple sclerosis in populations of European ancestry, but the associations are not always reproducible in other populations due to admixture and different linkage disequilibrium patterns obscuring true association signals. Objective: Our aim was to identify genetic predictors of multiple sclerosis in three ethnically homogenous populations from the Volga-Ural region of Russian Federation. Methods: In the largest to date study of multiple sclerosis in Russian population, involving 2048 participants from the Republic of Bashkortostan, Russian Federation (641 patients with multiple sclerosis and 1407 unaffected individuals), we performed replication analysis of previously identified genome-wide signals for multiple sclerosis. Associations were tested using logistic regression analysis under additive genetic model adjusted for sex. Meta-analysis of the study results in three populations was performed under fixed effects and random effects models. Results: We demonstrate the association with multiple sclerosis of the five variants (INAVA rs7522462, EOMES rs11129295, C6orf10 rs3129934, CD86 rs9282641, and GPR65 rs2119704). The strongest association (OR = 2.16, CI:1.85-2.74, P = 2.53x10(-13)) was detected for rs3129934 polymorphism in the major histocompatibility region. Multilocus analysis has revealed 322 and 27 allelic patterns associated with multiple sclerosis in women and men, respectively. In women, the highest risk of MS was conferred by C6orf10 rs3129934*T/T + STAT3 rs744166*T combination (OR = 11.87), in men - by C6orf10 rs3129934*T + EOMES rs11129295*C + RPS6KB1 rs180515*C combination (OR = 3.25). Conclusion: We confirm five associations with multiple sclerosis previously reported in genome-wide scans in Europeans in three ethnic groups from the Volga-Ural region of Russia.

For the first time in the ethnic group of Abkhazians, the association analysis of polymorphic DNA-markers of the antioxidant genes CAT (rs1001179), MSRA (rs10098474), GPX1 (rs1050450), GSR (rs1002149), GSTP1 (rs1695), SOD1 (rs2070424), SOD2 (rs4880), PON1 (rs662), PON2 (rs7493) with age was performed. Using ROC-analysis and logistic regression, it was found that the spectrum of alleles and genotypes frequencies of PON1 and GSTP1 genes polymorphic markers change throughout the studied age period (21-107 years old); the distribution of allele and genotype frequencies of CAT and SOD2 genes polymorphic markers changes within the age of 60 years. Multilocus genetic markers of longevity were determined by the Monte Carlo Markov chain method. Among persons in the age range 60-107 years, the frequency of observation of the patterns GSTP1*G/G+PON1*G (OR=6,59, PFDR=0,018) and GSTP1*G/G+SOD1*A (OR=3,4, PFDR=0,041) is statistically significantly increased; the GSTP1*A allele in various combinations with the PON1*A, PON2*C and CAT*C alleles are less common (OR=0,3, PFDR<0,05).

Objectives. To carry out a replicative analysis of the associations with multiple sclerosis of genetic markers of autoimmune diseases identified as a result of genome-wide studies in ethnically homogeneous groups of Russians and Tatars living in the Republic of Bashkortostan. Materials and methods. A group of 1724 people (547 patients with multiple sclerosis, 1177 members of the control group) underwent genotyping using allele-specific PCR and PCT with restriction fragment length polymorphism analysis for polymorphic variants rs2069762 of the IL2 gene, rs759648 of the PVT1 gene, rs1800682 of the FAS gene, and rs12708716 of the CLEC16A gene. Associations of these genetic markers with multiple sclerosis were studied using the PLINK program by logistical regression using an additive genetic model with sex as a covariate. Results. In the Tatar group we found an association between the rs759648*C of PVT1 with multiple sclerosis (OR = 1.42, p = 0.023). Meta-analysis of the results in the two ethnic groups confirmed the association of the rs759648*C allele of PVT1 with the disease (a random effects model and a fixed effect model: OR = 1.29, p = 0.018). Conclusions. These data provide evidence of an association between the polymorphic variant rs759648 of PT1 and multiple sclerosis in the Russian and Tatar populations living in the Republic of Bashkortostan. No associations with the other polymorphic variants studied were found in these two ethnic groups.

Рассеянный склероз - это хроническое воспалительное демиелинизирующее заболевание центральной нервной системы многофакторной природы. Цель настоящего исследования состояла в проведении анализа ассоциаций с заболеванием полиморфных маркеров в генах, продукты которых предположительно участвуют в патогенезе заболевания, а также идентифицированных в результате проведения полногеномных исследований, в популяциях башкир, русских и татар, проживающих в Республике Башкортостан. В результате проведенного исследования нами получены данные, подтверждающие ассоциацию с рассеянным склерозом генов, продукты которых участвуют в развитии аутоиммунного воспаления центральной нервной системы. Multiple sclerosis is a chronic inflammatory demyelinating disorder of central nervous system of multifactorial origin. Genome-wide association studies identified 700 polymorphic genetic variants associated with multiple sclerosis. The aim of the current study was to analyse associations of the polymorphic markers in genes involved in the pathogenesis of the disease and those identified by genome-wide association studies in the populations of Bashkirs, Russians, and Tatars from the Republic of Bashkortostan (Russian Federation). The study group was comprised of 644 patients with multiple sclerosis and 1408 control group individuals. Genotyping of 35 polymorphic genetic markers was performed using allele-specific PCR and PCR followed by restriction fragment length polymorphism analysis. The data obtained in our study confirm the association with multiple sclerosis of the genes involved in the development of autoimmune inflammation in central nervous system.

For the first time, a study of genetic factors of longevity was conducted in the prevalent populations of 2511 residents of the Republic of Bashkortostan—Russians, Bashkirs, and Tatars. We investigated the polymorphic markers in the genes of the antioxidant defense enzymes— SOD1 (rs2070424), SOD2 (rs4880), and SOD3 (rs1799895). We detected ethnicity-specific patterns of the distribution of genotype frequencies between Bashkir and Russian groups (rs2070424 of the SOD1 gene, P = 0.003), as well as between Tatars and the groups of Russians and Bashkirs (rs4880 of the SOD2 gene, P < 0.001 and 0.035, respectively). We found associations of the polymorphic markers in SOD family genes with age. Among Russians, the chances to attain longevity were higher in the SOD1 *А/А genotype carriers (OR = 1.025, P = 0.001) and lower in those with the SOD1 *А/G (OR = 0.975, Р = 0.001) and SOD2 *А/А (OR = 0.985, Р = 0.002) genotypes. Among Tatars, we observed a decrease in the SOD2 *A/A (OR = 0.989, Р = 0.029) and SOD2 *V/V (OR = 0.985, Р < 0.001) genotype frequencies and an increase in the SOD2 *A/V genotype frequency (OR = 1.023, P < 0.001). The analysis of genotype and/or allelic combinations of the studied polymorphic loci revealed 12 patterns associated with longevity among Tatars. The SOD1 *А and SOD3 *С alleles were present in most of the identified combinations. The SOD2 rs4880 polymorphic marker was indicative of longevity: combinations including the SOD2 *V/V genotype were associated with lower chances of achieving longevity (OR ≤ 0.45, P FDR ≤ 0.0003), and combinations including the SOD2 *A/V genotype were associated with higher chances of achieving longevity (OR ≥ 2.92, P FDR ≤ 1.24 × 10 –6 ).

Myocardial infarction (MI) is a multifactorial polygenic disease. It develops because of the complex interaction between many environmental and genetic factors. In this investigation, we have studied associations of MI and rs1042034 (gene APOB), rs4420638 (gene APOC1) rs2070424 (gene SOD1) и rs662 (ген PON1) in an ethnically homogeneous group. The material for analysis was DNA samples of patients (365 men) with onset of MI at the age of 30 to 60 years and 292 essentially healthy men of the control group. The study revealed markers of increased risk of MI: SOD1*A/A, for men under 46 years of age (P=0.029, OR=1.96), APOC1*A/G (P=0.03, OR=2.01), for men over 48 years of age, and APOB*C/C (P=0.031OR=1.8).

Долголетие – сложный социально-биологический феномен, при котором человек достигает возраста, значительно превышающего средний популяционный показатель. Вероятность достижения долголетия зависит от совокупности экзогенных и эндогенных факторов. На молекулярно-генетическом уровне характер их взаимодействия определяют гены ферментов метаболизма кислорода и ксенобиотиков, выполняющих функцию защиты организма от токсичных действий внешних агентов. Одним из таких генов является ген параоксоназы 1. Аллели полиморфного маркера rs662 (192Q>R) гена PON1 ассоциированы с активностью фермента и развитием сложно-наследуемых заболеваний, ограничивающих продолжительность жизни. С целью поиска ассоциаций полиморфного маркера rs662 гена PON1 с долголетием в этнической группе башкир проведен сравнительный анализ частот аллелей и генотипов в разных возрастных группах; в общей группе выполнен поиск возрастной динамики частот генотипов с помощью регрессионного анализа. В этнической группе башкир аллели PON1*Q, PON1*R и генотипы PON1*Q/Q, PON1*Q/R, PON1*R/R обнаружены с частотами 69.96, 30.04, 48.7, 42.51, 5.76% соответственно. Полиморфный маркер rs662 гена PON1 оказался ассоциирован с возрастом: шанс достичь долголетия выше у носителей гетерозиготного генотипа PON1*Q/R (55-100 лет, OR=1.022, P=0.043).

Согласно одной из современных концепций старения и долголетия человека, причиной деградации эндогенных процессов является повышение нестабильности генома с возрастом. К факторам генетической нестабильности относятся транспозоны, или мобильные генетические элементы. В геноме человека широко распространены Alu-ретротранспозоны. Среди членов данного семейства AluYa5 и AluYb8 являются единственными, сохранившими транспозиционную активность. С целью проверки гипотезы о роли активности транспозонов AluYa5 и AluYb8 в старении и долголетии человека проведен сравнительный анализ уровня их экспрессии среди лиц разного возраста. В исследовании приняли участие 75 здоровых жителей Республики Башкортостан в возрасте от 21 до 97 лет. Общая выборка дифференцирована на возрастные группы: среднюю (38 человек, 21-39 лет), старческую (23 человека, 82-89 лет) и группу долгожителей (14 человек, 90-97 лет). Образцы РНК выделяли из лейкоцитов периферической венозной крови стандартным тризольным методом. Количественный анализ содержания AluYa5-РНК и AluYb8-РНК в лейкоцитах крови проводили методом ПЦР в реальном времени. Относительное количество мРНК определяли с помощью ΔΔCt-метода. Сравнение возрастных групп проводили с использованием однофакторного дисперсионного анализа Краскела-Уоллиса. В общей анализируемой выборке показатели относительного уровня экспрессии субсемейств AluYa5 и AluYb8 составили 0,84 и 0,80 отн. ед. соответственно. Согласно сравнительному анализу относительного уровня AluYa5-РНК и AluYb8-РНК в лейкоцитах крови людей, относящихся к трем возрастным группам, не наблюдается статистически значимых различий в экспрессии транспозонов семейств AluYa5 (Н=3,59, p=0,17) и AluYb8 (Н=2,65, p=0,27). После объединения групп лиц старческого возраста и долгожителей и сравнения их с лицами среднего возраста различия в показателях значений медианы также не достигли уровня статистической значимости (p=0,1). При этом у людей в возрасте 82 лет и старше наблюдается более низкий уровень экспрессии Alu-элементов субсемейства AluYa5 (0,75 против 2,69 отн. ед.) и AluYb8 (0,73 против 2,15 отн. ед.). Таким образом, в исследованной выборке не было обнаружено статистически значимых изменений относительного уровня экспрессии субсемейств AluYa5 и AluYb8 с возрастом. According to one of the modern concepts of human aging and longevity, the cause of the degradation of endogenous processes is the increased genome instability with age. Factors of genetic instability include transposons, or mobile genetic elements. Alu-retrotransposons are ubiquitous in the human genome. Among the members of this family, AluYa5 and AluYb8 are the only ones that retained transpositional activity. To test the hypothesis about the role of of AluYa5 and AluYb8 transposons activity in human aging and longevity, we performed the comparative analysis of their expression level among individuals of different ages. The study group included 75 healthy residents of the Republic of Bashkortostan aged between 21 and 97 years. The total sample was divided into the following groups according to age: middle-aged (38 people, 21-39 years old), elderly (23 people, 82-89 years old) and a group of long-livers (14 people, 90-97 years old). RNA samples were isolated from leukocytes of peripheral venous blood by the standard method using Trizol reagent. Quantitative analysis of AluYa5-RNA and AluYb8-RNA in blood leukocytes was performed by real-time PCR. The relative amount of mRNA was determined using the ΔΔCt method. Comparison of age groups was carried out using one-way analysis of variance with the Kruskal-Wallis test. In the total analyzed sample, the expression levels of AluYa5 and AluYb8 were 0.84 and 0.80 rel. units respectively. According to the results of comparative analysis of AluYa5-RNA and AluYb8-RNA in blood leukocytes of people from three age groups, there were no statistically significant differences in the expression of AluYa5 (H = 3.59, p = 0.17) and AluYb8 (H = 2.65, p = 0.27). After combining the groups of elderly people and long-livers and comparing them with middle-aged people, the differences in the median values also did not reach the level of statistical significance (p = 0.1). At the same time, people aged 82 and older demonstrated lower level of expression of AluYa5 subfamily (0.75 versus 2.69 rel. units) and AluYb8 subfamily (0.73 versus 2.15 rel. units). There were no statistically significant changes in the relative expression level of the AluYa5 and AluYb8 subfamilies with age in the study group.