Dysregulation of blood pressure significantly impacts the course and outcome of sepsis. The AGTR1 gene encodes angiotensin II receptor type 1, which affects the vascular tone and contributes to septic shock. Our study aims to define whether the AGTR1 polymorphism contributes to the course and outcome of sepsis. The study included patients (n=286) from three ICU (Intensive Care Unit) aged 18-92 years with sepsis. In all patients (n=286) with sepsis the incidence of septic shock differed depending on AGTR1 genotype: carriers of the AGTR1 TT genotype had a less incidence of septic shock compared with patients with the AGTR1 AT, AA genotypes (60% vs. 73%, P = 0.47, FET (Fisher`s Exact Test), OR=1.8, 95% CI: 1.1-3.3). In group of patients with diabetes mellitus (n=79), we also found differences in sepsis course and outcome based on the AGTR1 rs275651 genotypes. The subgroup of TT AGTR1 rs275651 genotype carriers demonstrated significantly lower mortality compared with TA, AA genotypes carriers (60 % vs. 88 %, Р=0,018, OR=5,0, 95% CI: 1,34 -18,9, FET). The incidence of septic shock was also less in AGTR1 TT genotype-carriers in subgroup patients with diabetes (57% vs 92%, P=0,02, OR=9,2, 95 % CI: 1.97-42.9, FET). We found an association of the functional polymorphism AGTR1 rs275651 with course and outcome of sepsis in ICU patients with diabetes mellitus: carriers of the more common TT genotype had less incidence of septic shock and lower mortality compared to carriers of the minor A allele.

Abdominal obesity connecting with polygenic hereditary defects is considered the initial event in metabolic syndrome (MS) development. The purpose of study was to analyse the frequency of adiponectin (ADIPOQ) and leptin (LEP) gene polymorphisms in patients with MS, and the association of MS’s symptoms with named genes’ polymorphism. DNA was isolated from the whole blood of 207 patients with MS and 100 healthy individuals (control group). Gene polymorphisms were determined by real-time polymerase chain reaction. The association of the GG genotype of -2548 A/G polymorphism of the LEP gene (rs7799039) with the risk of high blood pressure in patients with MS, and the genotype of GG polymorphism + 45 T/G of the ADIPOQ gene (rs2241766) with hyperglycemia as a leading link in the development of MS was established. The association of the allele G of polymorphism +45 T/G (rs2241766) of the ADIPOQ gene with the risk of metabolic syndrome was established (OR (95% CI) =5,6 (2,4-13,0)), as well as the association of the TG and GG genotypes of this genetic variant (OR (95% CI)=3.81 (1.79-8.09) and OR (95% CI)=10.0 (2.25-44.7).

Purpose: To investigate the associations of FTO, FABP2, PPARG gene polymorphism and body mass index in Kuzbass residents. Methods. The sample for the study was 395 men, residents of Kuzbass. The average age of the subjects was 59±7.7 years. According to the questionnaire data, all the examined men referred themselves to the Russian nationality. The frequency of obesity in this sample (body mass index>30) was 46.3%. Genotyping of 3 polymorphic variants: FTO (rs9939609), FABP2 (rs1799883), and PPARG (rs1801282) were performed using TaqMan PCR. The association of these polymorphic variants with obesity in various inheritance models was then tested. Results. The associations of FTO rs9939609 gene variants with obesity in the codominant (for genotype T/A OR = 2.05; CI: 1.26-3.33; p = 0.004; for genotype A/A (OR = 2.27; CI: 1.28-4.04; p = 0.004) and dominant inheritance models (OR = 2.12; CI: 1.34-3.34 p = 0.001) were detected. Other genes showed no significant association with obesity in our sample. Conclusion. The rs9939609 variant of the FTO gene is associated with an increased risk of obesity in Kuzbass residents.

The authors searched for genetic and epigenetic markers of osteoporosis associated with the development of fractures and low bone mineral density and also developed clinical and genetic models of OP in 701 and 476 male samples divided into comparison groups depending on the presence or absence of osteoporotic fractures and variation in the BMD in different skeletal parts. Using the technology of KASP-genotyping and nonparametric criteria of statistical analysis, polymorphic variants of the OPG gene, miR-146, as well as microRNA binding sites of the VDR, ZNF239 and FGF2 genes, and variants of the OPG gene, miR-196 and microRNA binding sites of FBOX5, SOX9, MMP1 and ZNF239 genes were found to be markers of fractures. We performed a polygenic risk assessment of 140 full genome-wide association replication (GWAS) DNA locus and established clinical and genetic models predicting OP risk with up to 90% efficiency, with a 6,6-fold increased risk of fracture. Analysis of the methylation profile of 4 genes using pyrosequencing revealed an association of RUNX2 gene hypomethylation with severe primary OP in a sample of women.

Background. Cystic fibrosis is an inherited autosomal recessive disease. Modern methods of DNA diagnosis open up the possibility to develop new effective methods of cystic fibrosis treatment, which are based on the impact on the molecular pathogenesis of the disease. A personalized approach to medical care for patients with cystic fibrosis, taking into account the genetic profile with the use of new DNA diagnostic technologies has raised the provision of medical and genetic counseling for burdened families to a new level and more effective treatment measures. Aim. Implementation of a program of modern molecular genetic diagnosis of inherited diseases using targeted therapy for cystic fibrosis. Methods. The data for the entire period of neonatal screening for cystic fibrosis in the Republic of Bashkortostan were analyzed. In 2021 there were 94 patients on the cystic fibrosis registry in the Republic of Bashkortostan. The majority of patients required DNA diagnosis to verify mutations in the CFTR gene with subsequent prescription of tragent therapy. In the first phase of the study, 94 patients with cystic fibrosis as well as their relatives were analyzed using multiplex ligase-dependent probe amplification (SALSA MLPA Probemix P091 CFTR, MRC Holland). In the next step, all exons of the CFTR gene were sequenced using next-generation sequencing (NGS) and Sanger methods. Results. A search for mutations was performed in 94 probands and 36 variants of changes in the CFTR gene with pathogenic significance for the development of cystic fibrosis were identified. No mutations were found on 2.6% of chromosomes, and mutations were identified on 97.3% of chromosomes. In 89 patients both mutations were identified (83.66%), in 5 probands only one heterozygous mutation was found. One patient was diagnosed as having сystic fibrosis. Conclusion. The development and implementation of the latest sequencing technologies have significantly expanded knowledge about the molecular mechanisms of cystic fibrosis, which contributes to the development of new approaches to therapy and new research.

Hemophilia B is an X-linked recessive disease associated with blood clotting disorder caused by mutations in the coagulation factor IX (F9) gene. Both point sequence changes and large structural rearrangements were described in the gene. The aim of this study was to determine the spectrum of mutations in the F9 gene in Russian patients with hemophilia B. A study of 59 unrelated families with a referral diagnosis of “hemophilia B” was conducted using Sanger sequencing, mass parallel sequencing, and quantitative MLPA analysis. Mutations were detected in 86.4% of cases. The prevalent among the identified pathogenic variants were missense mutations - 62.7%; nonsense mutations account for 19.6%, large deletions involving one or more exons - 3.9%, mutations with a reading frame shift - 5.9%, splice site mutations - 2%, mutations in the promoter- 5.9%, which generally corresponds to the literature data. Fourteen identified nucleotide variants have not been previously described. Most of them were missense mutations. Thus, we studied the spectrum of mutations in Russian patients with hemophilia B. Determining the type of mutation and its localization in the gene and protein will allow to predict the course of the disease and to choose the optimal treatment.

Achondroplasia is a congenital disease, the most common form of short-limb dwarfism, inherited by autosomal dominant type. The pathogenesis is based on a disruption of endochondral ossification. Patients exhibit short stature with shortening of the proximal parts of the limbs, facial abnormalities. Hypochondroplasia is much milder. Both disorders are causes by mutations in the FGFR3 gene. Achondroplasia is caused in 99% of cases by mutation c.1138G>A/C (p.Gly380Arg), 60% of hypochondroplasia is caused by variant c.1620C>A/G (p.Asn540Lys). The mutations leading to achondroplasia cause constitutive activation of FGFR3. Due to orthopedic and neurological complications, it is important to treat achondroplasia. Pharmacological treatment of the disease is being developed. Molecular genetic diagnosis of achondroplasia is relevant for the timely initiation of treatment. The paper presents the first results of selective screening program FSBI «RCMG» for the presence of frequent pathogenic variants in the FGFR3 gene. Based on the material of 270 patients the informativeness of the program was 70%. The share of the most frequent mutation р.Gly380Arg (c.1138G>A/С) account for 85.7%. A variant of р.Asn540Lys (c.1620C>A/G) was detected in 13.7% of cases.

Background. NGS-based exome sequencing, accepted as an outsourced service since 2016, is increasingly being used in the routine clinical practice of our medical genetic counseling centre. Evaluation of the diagnostic capabilities of exome sequencing is necessary to substantiate the effectiveness of NGS in situ in regional clinical settings. Aim: to evaluate the effectiveness of diagnosing genetic diseases in children in Khanty-Mansi region using exome sequencing. Methods. Exome sequencing by NGS, validation of NGS data by Sanger sequencing, family segregation analysis by Sanger sequencing. Results. Molecular diagnoses were reported in 27% (27/100) of children, in line with data published elsewhere. The diagnoses were more often diagnosed in persons with anomalies of the nervous, skeletal and respiratory systems. Predominantly autosomal dominant diseases were identified, among which the most common were neurofibromatosis, osteogenesis imperfecta, achondroplasia. Family segregation revealed 5 new variants in the TSC1, KAT6A, ANKRD26, ARID1B, RPGRIP1 genes.

In our study, we searched for pathogenic changes in target genes in 62 patients with osteogenesis imperfecta from 52 families from the Republic of Bashkortostan using NGS technology. The study revealed 29 mutations in 4 genes (COL1A1, COL1A2, IFITM5, P3H1) responsible for the development of OI in 42 patients from 32 families. Additionally, heterozygous mutations in the CLCN7, ALOX12B, PLEKHM1, ERCC4, ARSB, PTH1R, and TGFB1 genes were found in 7 patients with low-traumatic fractures that are not involved in the pathogenesis of OI.

Duchenne/Becker muscular dystrophy (DMD/DMB) is the most common form of muscular dystrophy, accounting for over 50% of all cases. Since 01.10.2018, a program of selective screening for DMD/DMB has been conducted on the basis of the DNA diagnostics laboratory of the Research Centre of Medical Genetics, aimed at early detection of patients. The main inclusion criteria are male gender, an established clinical diagnosis of DMD or DMB, or a significant increase in the level of creatine phosphokinase (>2000 U/l). At the first stage of screening, patients are scanned for extended deletions and duplications in the DMD gene using multiplex ligase-dependent probe amplification. The second stage is the search for small mutations using a custom NGS panel. In a screening of 1071 unrelated probands with a referral Duchenne/Becker diagnosis, pathogenic variants in the DMD gene were found in 818 boys (in 75,3% of cases). Also, in 96 (8.9%) cases, other genetic causes of progressive muscular dystrophies were identified.

Rubinstein-Taybi syndrome (RTS) - rare hereditary disorder characterized by intellectual disability and growth retardation in conjunction with specific craniofacial and skeletal features and a wide range of multiple congenital anomalies. To date, mutations in two genes: CREBBP and EP300 can be discovered in about 60% of clinically identified patients with RTS. We herein report the result of molecular analysis in a cohort of Russian RTS patients. Despite the fact that we cannot confirm the diagnosis of RTS in 40% of causes, there are possibility for a better understanding of causative molecular mechanisms and improving the diagnostic process.

A study was carried out to identify the spectrum and frequency of pathogenic changes in the PIK3CA gene in the tumor tissue preparations of 60 women with hormone-dependent HER2-negative breast cancer from the Republic of Bashkortostan using the allele-specific polymerase chain reaction. As a result 32 pathogenic variants in the PIK3CA gene were identified: 18 mutations in exon 9 and 14 mutations in exon 20. The most frequent mutation was H1047R (p.His1047Arg(c.3140A>G)), which accounted for 21.6%. The second most frequent mutation was E545K (p.Glu545Lys (c.1633G>A)), which was 20%. E542K (p.Glu542Lys(c.1624G>A)), E545A (p.Glu545Ala(c.1634A>C)) mutations in exon 9 and H1047L (p.His1047Leu(c.3140A>T)) in exon 20 were identified with a frequency of 8.3%, 1.6% and 1.6% respectively. The results show a high frequency of somatic mutations in the PIK3CA gene (50%), indicating the need to identify pathogenic changes for adequate therapy of breast cancer.

The reason of SPG47 is homozygous and compound-heterozygous mutations in AP4B1 gene. SPG47 is characterized by microcephaly, corpus callosum hypoplasia, neonatal muscular hypotension, followed by spasticity and severe mental retardation. Variant c.1160_1161del was identified in six patients from unrelated, non-consanguineous families. Was performed haplotype analysis AP4B1 gene for four from six patients on chromosomes with this variant. As a result, there wasn’t found the only haplotype linked to this variant. Therefore, this variant can be a “hot spot” mutation, or could have spread as a result the founder effect.

Rare clinical case of Tay-Sachs disease (TSD) with late clinical manifestation in a 33-year-old female of mixed ethnicity (Russian, Ukrainian, Mordovian, Udmurt) with symptoms of spinal amyotrophy and spinocerebellar ataxia is presented. Targeted NGS of coding regions of 300 genes clinically significant for the development of hereditary neuromuscular diseases, including gene HEXA, was carried out. Two pathogenic variants in the HEXA gene have been identified - NM_000520.6(HEXA): c.2T>C (p.Met1Thr) and c.805G>A (p.Gly269Ser). Both variants were validated by Sanger sequencing. It is well known that Gly269Ser mutation associated with the adult onset of symptoms. The phenotypes of patients with late manifestation of TSD differ significantly from infant and juvenile forms, are less known to clinicians and are worse diagnosed. The authors hope that the description of this case will complement the missing information and help in the diagnosis of TSD with late clinical manifestation.

Cystic fibrosis is one of the most common genetic diseases not only in Russia but in the world, it dramatically reduces lifespan and quality of life in the absence of effective treatment. In a time of cystic fibrosis precision therapy, the effective treatment is based on the exact genotype determination. However, the diagnostic search should not always be finished when two pathogenic variants in CFTR gene are detected. In recent years there has been an explosive growth of research studies dedicated to, so-called, complex alleles, - the presence of two or more variants in the cis-position, which may affect in a dramatic way a therapeutic response. The study was aimed to determine the frequency of a complex allele p.[L467F;F508del], that affects the response to lumacaftor/ivacaftor (Orkambi) in cystic fibrosis Russian patients, being homozygous for F508del .