Over the past decade, circulating small non-coding ribonucleic acid molecules (microRNAs) have demonstrated their potential as minimally invasive diagnostic and prognostic biomarkers of various diseases. Standardization of preanalytical and analytical factors, including collection, processing and storage of biosamples, plays a significant role in the reliability and reproducibility of circulating microRNA quantification. To date, there is no consensus regarding the data normalization used in the analysis of circulating microRNA expression. The review aim is to consider modern original papers on various storage conditions of biobanked plasma and serum samples with subsequent isolation of circulating microRNAs for analysis.

Circulating microribonucleic acids (microRNAs) are promising biomarkers of various diseases, but their clinical laboratory use requires highly sensitive, reproducible, reliable and sustainable methods for their accurate plasma and serum quantification. The preanalytical phase of studies conducted using biospecimens consists of their collection, processing, storage and transportation. Preanalytical conditions remain the main distorting factors in microRNA studies, and standardization of these conditions, carried out in biobanks, can improve the reproducibility of results and their comparison. The review aim is to consider the main contemporary original studies on preanalytical factors, which are an important source of variability in studies on circulating microRNAs at the stages from blood collection to plasma or serum production.

Coronary collateral circulation (CCC) above the coronary chronic total occlusion is an alternative blood supply to the ischemic myocardium and increases survival among patients with coronary artery disease. In this regard, identification of novel markers associated with the CCC severity has diagnostic potential for patient stratification. It has been shown that circulating microRNAs play an important role in almost all cardiovascular aspects, including the association of some microRNAs with the CCC severity. The aim of this review is to consider the main modern studies on association of circulating microRNAs and CCC severity in coronary chronic total occlusion in patients with coronary artery disease, followed by a functional analysis of the identified microRNAs.

Aim. To validate and evaluate the accuracy of 4 genetic risk scores (GRSs) for hypertension (HTN), previously created on European samples, on a population sample of the Ivanovo Oblast.Material and methods. For genetic analysis, targeted next-generation sequencing was used on a sample of the Central Russia (n=1682) based on the biobank collection. Four GRSs associated with HTN, previously developed for the European population, were selected for validation. The coefficient of determination and the area under the ROC curve were used as quality metrics for regression models. Additional validation was carried out to include all nucleotide sequence variants, regardless of linkage disequilibrium level. A combined GRS was compiled based on coefficients from individual GRSs using the clumping + thresholding (C+T) method.Results. The study demonstrated that the predictive value of previously developed GRSs when used for Central Russian population is lower than in the original studies. The proportion of explained variance was 0,5-0,8%. The best predictive ability (proportion of explained variance — 2,5%) was demonstrated using previously developed GRSs (Evangelou E, et al., 2018), which includes the largest number of nucleotide sequence variants (n=852).Conclusion. GRSs for HTN, developed on European samples, is not recommended for Russian population without preliminary validation. To create original GRSs, combining statistical parameters (β-coefficients and p-value) from different GRS is not recommended.

Aim. To investigate the association of 10 circulating plasma microRNAs with collateral flow degree in chronic total occlusion (CTO) patients with coronary artery disease (CAD).Materials and methods. Plasma expression levels of 10 circulating miRNAs were measured by real-time PCR using Taqman technology in a sample of 43 subjects. The study included patients with CAD and CTO with good (n=13) or poor (n=10) coronary collateral circulation (CCC) based on Rentrop classification and a control group of patients without significant coronary stenosis (n=20).Results. Significant differences in expression levels were found for 7 circulating miRNAs in patients with CTO and good CCC and for 5 microRNAs in the combined group of patients with CTO compared to the control group. Among the 7 microRNAs, decreased expression of hsa-miR-126-5p, hsa-miR-146a-5p, hsa-miR-155-5p, hsa-miR-15b-5p, hsa-miR-21-5p, hsa-miR-23a-3p and increased expression of hsa-miR-451a were detected. For the first time, we showed that the level of 2 microRNAs (hsa-miR-23a-3p, hsa-miR-21-5p) is significantly reduced and the level of hsa-miR-451a is increased in patients with CAD with good CCC.Conclusion. Plasma microRNAs with significant differences obtained can be used for further studies on a larger sample size as candidate biomarkers for assessing the severity of ССС in the presence of CTO.

Aim. To study genetic causes of decreased low-density lipoprotein cholesterol (LDL-C) in Russian patients.Material and methods. The study included the following Epidemiology of Cardiovascular Diseases and their Risk Factors in Regions of Russian Federation (ESSE-RF) participants: individuals with LDL-C<5th percentile, taking into account sex and age (n=52), who underwent targeted sequencing of protein-coding regions of 6 genes (APOB, PCSK9, MTTP, ANGPTL3, SAR1B, APOC3) and determination of the genetic risk score (GRS) for hypercholesterolemia; and a representative sample of the Ivanovo region population (ESSEIvanovo, n=1667), for which only GRS was determined. Genetic testing was performed using next generation sequencing.Results. In 10 (19,2%) of 52 participants with decreased LDL-C levels, the following rare variants potentially associated with hypocholesterolemia were identified: 8 — leading to a premature termination codon in the APOB gene, 1 — leading to a premature termination codon in the APOC3 gene and 1 missense variant in the PCSK9 gene. Of the 10 identified variants, 6 are described by us for the first time. GRS in the LDL-C group (0,27±0,25) was significantly lower than in the ESSE-Ivanovo population sample (0,43±0,27) (p=4,7×10-06).Conclusion. Genetic reasons explain decreased LDL-C levels (<5th percentile) in 32,7% of patients, of which only monogenic variants were identified in 13,5%, a combination of monogenic and polygenic hypocholesterolemia — in 5,7%, and polygenic hypocholesterolemia — in 13,5%.

Aim. To validate and evaluate the accuracy of 14 genetic risk scores (GRSs) for type 2 diabetes (T2D), created earlier in other countries, using a Russian population sample from the biobank of the National Medical Research Center for Therapy and Preventive Medicine. Material and methods. For genetic analysis, next generation sequencing data was used on a sample from the Russian population (n=1165) based on the biobank collection. The study included 14 GRSs associated with T2D. Results. The study demonstrated that the predictive power of 12 out of 14 GRSs for T2D was replicated in the Russian population. As quality metrics, we used the area under the ROC curve, which for models including only GRS varied from 54,49 to 59,46%, and for models including GRS, sex and age — from 77,56 to 78,75%. Conclusion. For the first time in Russia, a study of 14 T2D GRSs developed on other populations was conducted. Twelve GRSs have been validated and can be used in the future to improve risk prediction and prevention of T2D in Russia.

Представлен клинический случай пациентки с семейной гиперхолестеринемией, с выраженным повышением уровня холестерина липопротеидов низкой плотности (ХС-ЛПНП) (в анамнезе до 10,57 ммоль/л) и двумя гетерозиготными вероятно-патогенными вариантами в двух генах LDLR (ENSP00000454071.1:p.Gly592Glu) и APOB (ENSP00000233242.1:p.Arg3527Gln). На фоне соблюдения строгой гиполипидемической диеты с ограничением холестерина и животных жиров, приема трёхкомпонентной гиполипидемической терапии, состоящей из аторвастатина 40 мг/сут, эзетимиба 10 мг/сут и алирокумаба 150 мг подкожно 1 раз в две недели, удалось снизить уровень ХС-ЛПНП с 10,57 ммоль/л до 3,29 ммоль/л (- 68,9%). Недостижение целевого уровня ХС-ЛПНП у пациентки связано, во-первых, с исходным крайне высоким значением, а во-вторых, по-видимому, со сниженной эффективностью гиполипидемической терапии при сложных генетических нарушениях.

Aim . To search associations for lipid profile parameters (lowand highdensity lipoprotein cholesterol levels, triglycerides and total cholesterol) in population samples from two Russian regions and make a replication analysis of a previously published genome-wide association study (GWA study, GWAS) for residents of three other Russian regions. Material and methods . The study included representative samples from the Vologda (n=689) and Ivanovo (n=1675) regions collected for the Epidemiology of Cardiovascular Diseases and their Risk Factors in Regions of Russian Federation (ESSE-RF) study. We assessed lipid profile parameters and performed a targeted sequencing. A linear regression model adjusted for sex, age, and statin use was used to assess the associations of genomic variants with lipid profiles. The work replicated the results of a study by Usoltsev D, et al., 2023, carried out on population samples of individuals from St.Petersburg, Orenburg and Samara regions. Results. We identified variants for which associations with lipid parameters had previously been identified in a Russian sample. The proportion of replicated variants was 89% and 92% for the samples from the Vologda and Ivanovo regions, respectively. The directions of effects of all replicated variants in the previously published study (samples from the Orenburg and Samara regions and St. Petersburg) and in both studied samples (samples from the Ivanovo and Vologda regions) coincide. Conclusion. The results of the search for associations with lipid parameters in different Russian samples are consistent with each other.

Aim. To validate and evaluate the accuracy of 15 genetic risk scores (GRSs) for obesity, created in populations of European origin in the sample of two European Russia regions. Material and methods. Genetic testing has been performed using next generation sequencing on a sample from the Russian population (n=1179). The study included 15 GRS associated with body mass index (BMI) or waist-to-hip ratio adjusted for BMI (WHRadjBMI). Results. The predictive power of 8 out of 9 GRSs for obesity based on BMI remains the same for the Russian population. The predictive power of 6 GRSs for obesity based on WHRadjBMI is lower in the Russian population than in the reference sample. GRS reproducibility increases with the size of initial samples and number of variants included in the GRS increase. The use of GRSs for obesity based on BMI in the Russian population created on European populations is justified. Conclusion. For the first time in Russia, 15 obesity GRSs developed in European populations have been validated. The data obtained on the effectiveness of the considered GRS can be used in the future to improve the obesity prediction and prevention in Russia.

Aim. To evaluate the information content of genetic risk scores (GRSs) for coronary artery disease (CAD), previously developed on European populations, in representatives of the Russian population. Material and methods . The work involved 1685 people from the ESSE-Ivanovo epidemiological study. CAD was verified in 3,1% of individuals. The coronary composite endpoint was assessed annually during 8-year follow-up. Next generation sequencing was performed using a targeted panel. Logistic regression analysis and area under the ROC curve (AUC) were used. Age, sex, and smoking status were taken into account in the multivariate model. Results. Of the 16 GRSs included in the analysis, only 2 GRSs demonstrated significance in the univariate analysis of association with CAD (highest AUC — 0,577). In a multivariate model, with an increase by 1 standard deviation (SD) for the 6 studied GRSs, a significant association with CAD was obtained — the odds ratio varied in the range of 1,31-1,47. The two GRSs demonstrated significant differences in the incidence of CAD between the groups corresponding to the upper and lower quintiles. Forty-five endpoints were registered. The risk ratio for the end point with an increase in GRS by 1 SD, taking into account cofactors, exceeded statistical significance for the 9 analyzed GRS and was in the range of 1,36-1,54. Conclusion. For the first time in Russia, 16 CAD GRSs, previously developed on European samples, was validated. The results were reproduced only for a few of the studied CAD SGRs.

Министерство здравоохранения Российской ФедерацииФедеральное государственное бюджетное учреждение «Национальный медицинский исследовательский центр терапии и профилактической медицины» Российское общество профилактики неинфекционных заболеваний «УТВЕРЖДАЮ» академик РАН, профессор, директор ФГБУ «НМИЦ ТПМ» Минздрава России, главный внештатный специалист по терапии и общей врачебной практике Минздрава России

Aim. To demonstrate two generations of a family with a progressive course of left ventricular non-compaction (LVNC) and the presence of a RBM20 gene variant. Material and methods . Based on the multicenter registry of patients with LVNC, a family with LVNC with a dilated phenotype was selected at the National Medical Research Center for Therapy and Preventive Medicine. Next generation sequencing was performed on a Nextseq 550 systen (Illumina, USA). For clinical interpretation, nucleotide sequence variants in the genes associated with LVNC development were selected according to the available literature data, with frequencies <0,5% in the gnomAD database. The identified variants were verified using Sanger sequencing on an Applied Biosystem 3500 Genetic Analyzer (Thermo Fisher Scientific, USA). Results. The article presents the results of clinical, paraclinical and molecular genetic studies of two generations of a family diagnosed with LVNC with a dilated phenotype and the progression of isolated LVNC to a dilated type. As a result of a molecular genetic study, all family members with the LVNC were found to have a likely pathogenic variant in the RBM20 NP_001127835.2:p.Pro638Leu (rs267607003) gene. RBM20 is a key splicing regulator that controls the processing of several important transcripts predominantly expressed in striated muscle, especially cardiac tissue. RBM20 gene variants can lead to disruption of splicing at several points and, as a result, to cardiomyopathy progression. Most known pathogenic RBM20 variants are associated with dilated cardiomyopathy; however, a number of studies have found RBM20 gene variants in patients with LVNC. The segregation of nucleotide sequence variant with symptoms in two generations testifies in favor of the association of the detected variant with LVNC development. Conclusion. Currently, the boundaries of the cardiomyopathy genetics are expanding. Pathogenic and likely pathogenic RBM20 gene variants are associated primarily with a dilated phenotype and a high risk of sudden cardiac death. The article presents the results of a survey of two generations of a family with LVNC and progressive myocardial remodeling.

Aim. To study the contribution of rare and low-frequency variants of ANGPTL3, ANGPTL4, APOA5, APOB, APOC2, APOC3, LDLR, PCSK9, LPL genes in assessing the risk of coronary artery disease (CAD) in a cohort of Russian patients with various cardiovascular risks.Material and methods. The study was conducted on a sample of participants in cohort and epidemiological studies (n=2405). Targeted enrichment of coding sequences and exon-intron regions of nine genes (ANGPTL3, ANGPTL4, APOA5, APOB, APOC2, APOC3, LDLR, PCSK9, LPL) was performed. Genetic diagnostics was carried out by next generation sequencing.Results. CAD was confirmed in 267 patients (11%). After genetic diagnosis, all patients were divided into three following groups: individuals with previously described genetic variants associated with elevated levels of low-density lipoprotein cholesterol (LDL-C) and/or triglycerides (TGs); individuals with genetic variants associated with reduced levels of LDL-C and/or TGs; individuals without genetic variants associated with LDL-C and/or TG levels, or with two or more variants with opposite effects on LDL-C and/or TG levels. Kaplan-Meier method revealed that the groups significantly differ in cumulative risk of CAD (p<0,001 for the log-rank test), the maximum risk was in group 1, and the minimum risk in group 2. When conducting the Cox regression, we found that in persons from group 1, the hazard ratio (HR) for CAD is 2,63 times higher (HR =2,63, 95% confidence interval (CI), 1,6-4,34; p><0,001), and in persons from group 2 lower by 1,88 times (HR =0,53, 95% CI, 0,3-0,98; p=0,042) compared with persons from group 3, adjusted for other CAD risk factors: sex, age, smoking, LDL-C and hypertension. Conclusion. Genetic testing in young patients makes it possible to identify individuals with an increased genetic risk of CAD and to focus preventive and therapeutic measures primarily for this category of patients. Keywords: coronary artery disease, cardiovascular diseases, low-density lipoprotein cholesterol, genetic testing. Relationships and Activities: none. 1National Medical Research Center for Therapy and Preventive Medicine, Moscow; 2Pirogov Russian National Research Medical University, Moscow; 3E. I. Chazov National Medical Research Center of Cardiology, Moscow; 4Lomonosov Moscow State University, Moscow; 5Moscow Institute of Physics and Technology, Dolgoprudny, Russia.><0,001 for the log-rank test), the maximum risk was in group 1, and the minimum risk in group 2. When conducting the Cox regression, we found that in persons from group 1, the hazard ratio (HR) for CAD is 2,63 times higher (HR =2,63, 95% confidence interval (CI), 1,6-4,34; p<0,001), and in persons from group 2 lower by 1,88 times (HR =0,53, 95% CI, 0,3-0,98; p=0,042) compared with persons from group 3, adjusted for other CAD risk factors: sex, age, smoking, LDL-C and hypertension.Conclusion. Genetic testing in young patients makes it possible to identify individuals with an increased genetic risk of CAD and to focus preventive and therapeutic measures primarily for this category of patients.

The growing prevalence of metabolic disorders creates an increasing demand for novel approaches to their prevention and therapy. Novel genetic diagnostic technologies are developed every year, which makes it possible to identify people who are at the highest genetic risk of diabetes, non-alcoholic fatty liver disease, and metabolic syndrome. Early intervention strategies can be used to prevent metabolic disorders in this group of people. Genetic risk scores (GRSs) are a powerful tool to identify people with a high genetic risk. Millions of genetic variants are analyzed in genome-wide association studies in order to combine them into GRSs. It has become possible to store and process such huge amounts of data with the help of biobanks, where biological samples are stored according to international standards. Genetic studies include more and more people every year that increases the predictive power of GRSs. It has already been demonstrated that the use of GRSs makes future preventive measures more effective. In the near future, GRSs are likely to become part of clinical guidelines so that they can be widely used to identify people at high risk for metabolic syndrome and its components.

Background: The identification of genetic factors that are simultaneously responsible for the predisposition to the development of cardiovascular diseases (CVD) and osteoporosis (OP) is important for the prevention of both conditions.Aim: The aim of this study is to evaluate three genetic risk scales (GRS) that previously showed an association with bone mineral density (BMD) and fracture risk, as well as to study the associations of these GRS with vascular wall pathology.Materials and methods: 250 female outpatients (aged 45 to 69) were enrolled into a cross-sectional study. The intima-media thickness (IMT), the presence and number of atherosclerotic plaques (AP) were studied using duplex scanning. Pulse wave velocity (PWV), augmentation index (AI) were measured by applanation tonometry. Coronary vessels calcium deposits were registered by multispiral computed tomography (MSCT) using the Agatston calcium index (CI). The BMD of the spine, hip neck (HN) and proximal hip (PH) was measured using double energy x-ray absorptiometry. Bone resorption marker type-1 collagen C-terminal telopeptide (CTx) was assessed solid-phase enzyme immunoassay. The genetic study included DNA extraction from whole blood samples. Targeted sequencing was performed on the Nextseq550 sequencer (Illumina, USA). Statistical analysis was carried out using the SAS software package for Windows, version 9.0 (SAS Institute Inc., USA).Results: The chance of detecting low bone mass increased more than 4 times at values of IMT ≥0.9 mm (OR=4.17; 95%CI [1.2–14.4], p<0.02), 2.4 times in the presence of AP in the carotid arteries (OR=2.45; 95%CI [1.12–4.88], p><0.05), by 6.7 times with an Agatstone CI ≥ 100 units (OR=6.68; 95%CI [1.56–28.7], p><0.001), 1.4 times (OR=1.43; 95%CI [0.56–3.68], p><0.438) with a PWV ≥10 m/s, 1.2 times (OR=1.2; 95%CI [0.601–2.43], p><0.60) with increased AI ≥ 27%. According to multivariate linear regression analysis (adjusted for age, duration of postmenopause, marker of bone resorption CTx), a significant association of all GRS with BMD in all parts of the skeleton was revealed. Both univariate and multivariate regression models adjusted for several covariants (age, total cholesterol, systolic blood pressure) showed a reliable association of GRS62 with the presence of plaques and GRS63 — with coronary artery CI. Conclusion: The results of the study demonstrated the association of polygenic genetic risk of GRS-based OP with BMD and vascular wall status indicators in women in the peri and postmenopausal periods.>< 0.02), 2.4 times in the presence of AP in the carotid arteries (OR=2.45; 95%CI [1.12–4.88], p< 0.05), by 6.7 times with an Agatstone CI ≥ 100 units (OR=6.68; 95%CI [1.56–28.7], p< 0.001), 1.4 times (OR=1.43; 95%CI [0.56–3.68], p< 0.438) with a PWV ≥10 m/s, 1.2 times (OR=1.2; 95%CI [0.601–2.43], p<0.60) with increased AI ≥ 27%. According to multivariate linear regression analysis (adjusted for age, duration of postmenopause, marker of bone resorption CTx), a significant association of all GRS with BMD in all parts of the skeleton was revealed. Both univariate and multivariate regression models adjusted for several covariants (age, total cholesterol, systolic blood pressure) showed a reliable association of GRS62 with the presence of plaques and GRS63 — with coronary artery CI.>< 0.60) with increased AI ≥ 27%. According to multivariate linear regression analysis (adjusted for age, duration of postmenopause, marker of bone resorption CTx), a significant association of all GRS with BMD in all parts of the skeleton was revealed. Both univariate and multivariate regression models adjusted for several covariants (age, total cholesterol, systolic blood pressure) showed a reliable association of GRS62 with the presence of plaques and GRS63 — with coronary artery CI.Conclusion: The results of the study demonstrated the association of polygenic genetic risk of GRS-based OP with BMD and vascular wall status indicators in women in the peri and postmenopausal periods.

Представлен клинический случай пациентки с выраженным повышением уровня триглицеридов (в анамнезе до 26,28 ммоль/л) и ранее не описанными вероятно-патогенными вариантами в гене LPL:p.[(Asp202Asn)];[(Tyr233Cys)]. На фоне соблюдения строгой гиполипидемической диеты с ограничением растительных и животных жиров, приема фенофибрата 145 мг/сут и эзетимиба 10 мг/сут, удалось снизить уровень триглицеридов с 16,27 ммоль/л до 2,04-2,68 ммоль/л.

Osteoporosis is a chronic systemic disease of the skeleton, characterized by a decrease in bone mass and an impairment of bone microarchitecture, which can lead to a decrease in bone strength and an increase in the risk of minor trauma fractures. Osteoporosis is diagnosed on the basis of bone mineral density (BMD). BMD is characterized by high heritability that ranges according to various sources from 50 to 85%. As in the case of other complex traits, the most common approach to searching for genetic variants that affect BMD is a genome-wide association study. The lower effect size or frequency of a variant is, the larger the sample size is required to achieve statistically significant data on associations. Therefore, the studies involving hundreds of thousands of participants based on biobank data can identify the largest number of variants associated with BMD. In addition, biobank data are used in the development of genetic risk scores for osteoporosis that can be used both in combination with existing prognosis algorithms and independently of them. The aim of this review was to present the most significant studies of osteoporosis genetics, including those based on biobank data and genome-wide association studies, as well as studies on the genetic risk scores and the contribution of rare variants.