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Distribution of alleles and genotypes of the G10976A (rs6046) variant of the F7 gene in ethnic groups living in the Arkhangelsk Region
Background. The existing differences between the gene pools of the peoples of Russia determine the importance of conducting molecular genetic population studies of various genes that determine the determination of various pathological conditions. It is known that the frequency of F7 G10976A (rs6046) allele carriers varies significantly in different global populations. As a result of a single nucleotide substitution in the VII factor gene, the synthesis of proconvertin protein, which is an important component of the second phase of hemostasis, is reduced. Heterozygous carriers of this gene have a protective effect on atherosclerotic processes, while homozygous carriers of the mutation may experience its pathological effects in the form of mild to severe hemorrhagic syndrome. Aim: analysis of the distribution of allele and genotype frequencies of the G10976A (rs6046) variant of the F7 gene in practically healthy Russians and Indians, volunteers living in the Arkhangelsk Region. Methods. The study was conducted on samples of ethnic Russians who were natives of the Arkhangelsk Region (n=318) and ethnic Indians who were temporarily residing in the city of Arkhangelsk (n=213). A molecular genetic study of the G10976A single nucleotide substitution in the F7 gene was conducted using real-time PCR, followed by a comparative analysis of the allele frequency distribution of the studied genetic variant in the study groups with the frequencies for various global population groups. Results. In the group of Russian volunteers, the frequency of allele carriage was as follows: G – 84.7%, A – 15.3%. The frequency of genotypes was as follows: GG – 71.8%, GA – 25.9%, AA – 2.3%. In the group of Indians, the frequency of the G allele was 68.8%, and the frequency of the A allele was 31.2%; the frequencies of genotypes were as follows: GG – 47.3%, GA – 43.0%, AA – 9.7%. Statistically significant differences in the frequency of mutant allele A carriage were revealed between the analyzed groups (p<0.001). When conducting a pairwise comparison of the group of Russians, natives of the Arkhangelsk region, with other global populations, the results were correlated with the data obtained for the European population (p=0.244), in the group of Indians temporarily residing in the territory of Arkhangelsk, – with the data for the South Asian population (p=0.124).
Dynamics of the epigenetic index of stem cells as a prognostic factor for metastasis-free survival after neoadjuvant chemotherapy for breast cancer
Intratumoral heterogeneity, particularly the subpopulation of cancer stem cells (CSCs), is associated with therapy resistance and metastasis. The persistence of a small fraction of CSCs after neoadjuvant chemotherapy (NAC) underscores the need to evaluate the prognostic potential of the dynamic epigenetic stem cell index (ESCI) in the context of metastasis-free survival in patients with luminal B subtype breast cancer. Based on bisulfite sequencing data from public repositories, we constructed a reference epigenetic atlas comprising 88 samples of various cell and tissue types. Using Shannon entropy and the Wilcoxon test, 45 highly variable CpG pairs were selected and applied for epigenetic deconvolution. Statistical analysis and deconvolution were performed on 19 paired luminal B breast cancer samples collected before and after NAC. We found that a high baseline ESCI prior to treatment was associated with 10-year metastasis-free survival (p < 0.05). Following NAC, a significant reduction in ESCI was observed in the non-metastatic group, whereas the metastatic group retained baseline ESCI levels. Importantly, the dynamic reduction of ESCI in non-metastatic samples correlated significantly with improved metastasis-free survival (p < 0.05). Thus, ESCI dynamics determined by epigenetic deconvolution may serve as a promising prognostic and predictive biomarker for assessing the efficacy and outcomes of NAC in patients with luminal B breast cancer.
The frequency of variants of the MEFV gene responsible for the development of periodic disease in Dagestan residents
Familial Mediterranean fever (FMF) has a certain ethnic specificity. Some ethnic groups inhabiting the Republic of Dagestan have a common origin with the Mediterranean populations in which FMF is widespread. A survey of 390 representatives of the indigenous ethnic groups of Dagestan was conducted, and the frequencies of variants in the MEFV gene were determined.
Study of the effect of targeted therapy on the restoration of the functional CFTR channel in a patient with the E403D/CFTRdele2,3 genotype
Background. Cystic fibrosis (CF) is a systemic hereditary disease caused by a mutation in the CFTR gene and characterized by damage to the glands of external secretion, severe respiratory dysfunction. It is believed that with a «severe» genotype, clinical manifestations are more pronounced and manifest early. Variant E403D is included in the list of variants sensitive to CFTR modulators (ivacaftor-tezacaftorelexacaftor). However, its use in clinical practice in a patient – a carrier of this variant E403D, did not lead to the expected effect. Methods. The medical history of a patient with CF with the genotype E403D/CFTRdele2,3 was analyzed. Biopsy material from the rectum was used to intestinal current measurement (ICM) and forskolin-induced swelling assay in intestinal organoids. DNA for sequencing was isolated from venous blood leukocytes. Results. The patient (11-year-old girl) with the E403D/CFTRdele2.3 genotype was diagnosed based on neonatal screening. The ICM method revealed reduced CFTR channel function before the start of CFTR modulator therapy. According to the forskolininduced swelling assay in intestinal organoids, it was shown that the amount of functional protein on the apical membrane of the intestinal epithelium did not increase under the action of either the potentiator or the corrector in the patient. Clinically, no positive dynamics were obtained from taking the drug for 1.5 years. Conclusion. For the first time, an analysis of the functional state of the chloride channel of a patient with the E403D/CFTRdele2,3 genotype was performed using the ICM method before therapy and during CFTR modulator therapy. A relationship was shown between severe manifestations of the disease and the absence of CFTR chloride channel function, both according to the sweat test and the absence of a response to forskolin during the ICM method. Forskolin-induced swelling of intestinal organoids showed that therapy with the tested CFTR modulators cannot be recommended for a patient with the E403D variant in a compound heterozygous state with a class I or VII variant.
Molecular Dynamics of the Mutant GABA Transporter GAT-1: contribution to the Pathogenesis of Schizophrenia
Background. It is well established that the GABAergic system, which plays a crucial role in both activating and inhibiting neuronal communication, is involved in the pathogenesis of schizophrenia. Mutations in the structure of GABA transporters may represent a key process in the development of this disorder. Consequently, developing in silico models of protein transporter dynamics in the central nervous system could help elucidate their functional significance in the progression of the disease. Aim: to model the structure and dynamics of the GAT-1 transporter within the neuronal membrane, taking into account the known schizophrenia-associated mutations, in order to refine our understanding of their functional relevance. Methods. The molecular dynamics of both mutant and wild-type GABA transporter GAT-1 was simulated using GROMACS. The structures were based on PDB (7Y7V), with three missense substitutions (A93T, R211C, W495L) introduced in PyMOL. A membrane system was constructed via CHARMM-GUI, followed by solvation and ionization. Energy minimization was carried out, and the system was subsequently simulated under isobaric-isothermal conditions (T = 300 K) using the Parrinello-Rahman barostat. Dynamics were analyzed through RMSD, RMSF, and radius of gyration (Rg). Results. Molecular dynamics simulations (100 ns) revealed that mutations in SLC6A1 lead to significant disruptions in both global and local stability of the GABA transporter. RMSD and RMSF analyses showed delayed stabilization of the mutant protein, increased fluctuations in key ion transport regions, and a potential loss of transport function. A slight increase in Rg in the mutant suggests preserved packing density but impaired binding capacity. Conclusion. These alterations may negatively impact GABA reuptake, which is associated with cognitive impairments and schizophrenia symptoms, highlighting the need for further investigation into the structure and function of the transporter in the pathogenesis of neuropsychiatric disorders.
Disorders of spermatogenesis in carriers of balanced chromosome translocations
Introduction. Carriers of balanced chromosome translocations have an increased risk of gametogenesis and fertility disorders, aneuploidy in gametes and congenital malformations in offspring. Among men with infertility, the frequency of balanced translocations exceeds the general population, however, semen parameters, as well as the causes of spermatogenic disorders, are poorly studied in such patients. Objective: to evaluate the effect of balanced chromosome translocations on spermatogenesis and semen parameters Methods. Male carriers of balanced chromosome translocations: Robertsonian (rob) (n=37) and autosome reciprocal translocations (n=38) were examined Standard semen analysis and quantitative karyological analysis of immature germ cells were performed. Results. Semen disorders, varying from moderate to severe forms, were detected in 92% of carriers of autosome reciprocal translocations carriers and in 95% of rob carriers. In rob carriers oligoasthenoteratozoospermia (OAT) was the most common. Patients with OAT showed partial arrest of spermatogenesis in meiotic prophase I. Conclusions. A difference in the structure of semen disorders was revealed in carriers of rob and reciprocal autosome translocations; oligozoospermia was the most common in patients with rob, indicating a partial arrest of spermatogenesis. Impaired fertility in male carriers of balanced autosome translocations with OAT is associated with a partial meiotic arrest, leading to decreased gamete production.
Experience of applying criteria for including pregnant women in the research project «Implementation of non-invasive prenatal screening in prenatal diagnosis of pregnant women»
The biomedical study «Implementation of non-invasive prenatal screening in prenatal diagnostics of pregnant women» is a stage of the research project «Study of the effectiveness of introducing non-invasive prenatal screening in examining women’s health», implemented in St. Petersburg since 2021. The effectiveness of non-invasive prenatal testing (NIPT), the results of which can influence pregnancy management tactics, requires constant monitoring and evaluation.
Clinical and genetic polymorphism of RASopathies in patients with left ventricular hypertrophy
RASopathies are a heterogeneous group of diseases caused by genetic alteration of the RAS-MAPK pathway, which are characterized by multisystem symptoms, including short stature, facial dysmorphia, and a wide range of cardiac manifestations. The aim of the study was to evaluate the spectrum of genetic causes and clinical manifestations of RASopathies in patients with left ventricular hypertrophy (LVH). We examined 180 probands with LVH which require surgical treatment, and found 19 rare variants in the genes encoding RASMAPK proteins in 18 probands (1-68 years). In 15 patients (4.5%), the diagnosis of RASopathy was confirmed by the detection of causative variants in the PTPN11, RIT1, LZTR1, MRAS, and HRAS genes. The primary diagnosis was Noonan syndrome in 6 patients, and hypertrophic cardiomyopathy in 9 patients. Two probands with RIT1 mutations had a macrosomal phenotype which was not typical for RASopathies. During the follow-up, 4 probands died, and 1 underwent a heart transplant. Three probands with LVH were found to have rare VUS, which contribution requires further study. Conclusions: despite the well-known phenotype, many patients with RASopathies have not been clinically diagnosed. The phenotypic spectrum of RIT1 mutations can be expanded with macrosomy. In the presence of LVH, patients with rasopathies have an earlier manifestation and age of surgery, and a worse prognosis compared to non-syndromal HCM patients.
Potential of molecular karyotyping using NGS
The early genetic diagnosis of newborns with congenital pathology is an important issue. The purpose of this work was to evaluate the possibility of detecting copy number variations (CNVs) using high-throughput/whole exome sequencing (NGS/WES). The study included 802 newborns with phenotypic features, 32 (4%) of them were suspected to have CNVs and 10 (1,2%) of them were suspected to have aneuploidies by WES. Validation of the CNVs was performed using molecular karyotyping (CMA). Aneuploidies were verified by Quantitative Fluorescence Polymerase Chain Reaction (QF-PCR). The results of the work showed a high reliability to detect the CNVs using WES.
Creating a predictive model for the probability of a patient having familial hypercholesterolemia
The results of a study aimed at developing a prognostic model for identifying familial hypercholesterolemia (FH), the most common monogenic lipid metabolism disorder, are presented. A comprehensive clinical and genetic analysis was performed on a sample of 466 patients. It was demonstrated that a combined assessment of low-density lipoprotein (LDL) levels and the atherogenic index allows for a highly accurate prediction of a molecularly confirmed diagnosis of FH. Based on the developed model, an online calculator was developed and implemented for cardiologists and geneticists to optimize the diagnostic pathway and patient selection for genetic testing. The proposed tool demonstrates high sensitivity and specificity in both children and adults.
Dystrophinopathies are a spectrum of X-linked muscular disorders associated with pathogenic/likely pathogenic variants in the dystrophin (DMD). Most often, these diseases affect men, while women are typically asymptomatic carriers and do not exhibit clinical symptoms. However, to this day, there has been an increasing number of reported cases of dystrophinopathy manifesting in women. In our study, we present a cohort of 30 women exhibiting clinical symptoms of dystrophinopathy, all of whom carry a heterozygous pathogenic variant in the dystrophin gene. In 20 cases, whole-genome sequencing was performed, ruling out other genetic variants that could account for similar phenotypic manifestations of muscular dystrophy. In three cases, a translocation was suspected based on genomic analysis, and a standard cytogenetic was subsequently conducted. In all other cases, no variants were identified that could explain the phenotype of muscular dystrophy, except for pathogenic variants in the DMD. It discusses the importance of mutation screening in the DMD for women presenting with muscular dystrophy symptoms and investigating carrier status in relatives of patients with Duchenne/ Becker muscular dystrophy.
Study of the role of the BDNF gene in the formation of differences in the level of state trait anxiety
The study is devoted to investigating the genetic predisposition to anxiety disorders, particularly the role of the BDNF (brain-derived neurotrophic factor) gene in the formation of state and trait anxiety. The work analyzed data from 333 mentally healthy students from the Republic of Bashkortostan. Methods of genetic analysis (PCR, fluorescence detection) and psychometric testing (Spielberger-Hanin questionnaire) were used. The results revealed significant associations of the polymorphic loci rs6265 and rs712442 of the BDNF gene with variations in anxiety levels. The rs6265A allele was associated with lower levels of trait anxiety (β = -1.748; p = 0.02), while the rs712442T allele was linked to higher levels of state anxiety (β = 1.27; p = 0.03). Additionally, gene-environment interactions between BDNF polymorphisms and the history of COVID-19 were found to influence state anxiety levels. Differences in anxiety were also identified based on gender, maternal age at childbirth, birth weight, antibiotic use, and hormone therapy. The findings emphasize the importance of a comprehensive approach to studying anxiety, considering both genetic and environmental factors.
Development and validation of a novel method for generating locus-specific DNA probes for the diagnosis of chromosomal aberrations
Introduction. Development of a domestic panel of DNA probes for detection of the most frequent chromosomal aberrations is a critical task to ensure independent and high quality molecular cytogenetic diagnostics in the Russian Federation. Objective. To develop a domestic panel of DNA probes for detection of chromosomal aberrations. Methods. The method of obtaining a panel of DNA probes for the detection of chromosomal aberrations included the development of probes using long-range PCR with the subsequent formation of DNA libraries for the diagnosed regions of the human genome and control loci. Results. The technology for obtaining DNA probes for FISH diagnostics and a panel of DNA probes from domestically produced components were developed.
Protein-Variant-Phenotype study of NBAS in the context of nonsense-mediated RNA decay proteins using AlphaFold
Pathogenic variants in the NBAS cause multisystem disorders, including SOPH syndrome and ILFS2. NBAS performs multiple cellular functions, which may explain the complexity of its clinical manifestations. Previously, we developed the Protein-Variant-Phenotype (PVP) approach using AlphaFold to analyze binding sites and their impact on phenotype. In this study, we used AlphaFold3 to investigate NBAS interactions with SMG proteins of the NMD complex to explore the potential contribution of disrupted protein interactions to the multisystem phenotype. We identified consensus binding sites between NBAS and SMG9 in the C-terminal region, which is associated with optic nerve atrophy. These findings suggest an alternative mechanism for NBAS involvement in NMD through auxiliary SMG proteins. Furthermore, this study is the first to associate C-terminal of NBAS with a specific phenotypic trait and cellular mechanism, providing a foundation for potential therapeutic approaches.
A multilevel model for assessing the risk of prereproductive mortality of offspring in human populations
The model imitating the processes (transmission in a series of generations) and variability (mutational and combinational) of genes that determine a multifactorial trait («pre-productive death of offspring») in a population. The model uses a normal distribution to estimate the probability of pre–productive death of offspring at various stages of the implementation of a genotype into a phenotype (a gene as a DNA reading chain – chromosome – organism – population). The model allows us to obtain prognostic estimates of the survival of a human population in a given number of generations, taking into account the segregation burden, the risk of new gene mutations, and other environmental factors.
Investigation of the Role of the Dopaminergic System in Pain Catastrophizing and Memory in a Nitroglycerin-Induced Migraine Model in Rats
Migraine is a complex neurological disorder accompanied by cognitive impairments, including memory problems and pain catastrophizing. One of the proposed mechanisms underlying these changes is the dopaminergic system, which plays a key role in cognitive processes. This study aimed to investigate the role of the dopaminergic system in pain catastrophizing and long-term memory in a nitroglycerin-induced migraine model in rats. The experiment utilized DAT-KO rats, including DAT-WT (n = 14) and DAT-HET (n = 14). A migraine-like phenotype was induced by an intraperitoneal injection of nitroglycerin (10 mg/kg). Pain catastrophizing was assessed using a conditioned place avoidance paradigm with a needle and von Frey filaments, while long-term memory was evaluated using the novel object recognition test. Data were analyzed using a two-way ANOVA. No significant differences in pain catastrophizing were found between the groups (p > 0.05). The discrimination index for memory also did not differ between genotypes and was not affected in the migraine model (p > 0.05). Thus, the study demonstrated that pain catastrophizing does not develop in the examined groups, and nitroglycerin administration does not affect memory processes in rats regardless of their dopaminergic status.
Molecular genetic analysis of Rubinstein–Taybi syndrome in Russia
Rubinstein-Taybi Syndrome (RTS) is a rare pathology, characterized by intellectual disability, developmental delays and a specific set of phenotypic features, with various anomalies in internal organs. The diagnostic algorithm includes detecting mutations in two main genes: CREBBP and EP300. This study presents the results of molecular-genetic analysis of 158 Russian patients with RTS: pathogenic and likely pathogenic variants were identified in 70 patients (42.4%), of which 62 (39%) were associated with the CREBBP gene and 4 cases (2%) with the EP300 gene. One patient was found to have a pathogenic variant in the SRCAP gene, which is associated with the development of a phenotypically similar condition known as Floating-Harbor Syndrome (FHS). There are opportunities for improvement in diagnostic principles.
HSP70 loci significantly increase the risk of hypertension in hospitalized patients with COVID-19
Cardiovascular diseases (CVD) significantly aggravate the severity and mortality of COVID-19. Heat shock proteins (HSPs) are known to be involved in the immune response and inflammation, key processes in the pathogenesis of both arterial hypertension (AH) and severe COVID-19. We aimed to investigate the association of polymorphic variants of the HSP70 family genes with the risk of AH in patients with severe COVID-19. Genotyping of 199 patients with severe COVID-19 (70 with AH and 129 without) was carried out by real-time PCR. Polymorphic variants of rs1043618 HSPA1A (risk allele C, OR = 2.0, 95% CI 1.24-3.22, p = 0,.03) and rs6457452 HSPA1B (risk allele T, OR = 2.99, 95% CI 1.32-6.78, p = 0.003) were associated with an increased risk of AH in patients with severe COVID-19. Analysis of ciseQTL effects found that rs1043618 HSPA1A and rs6457452 HSPA1B significantly affect the expression of histocompatibility complex genes, which can enhance inflammatory responses and contribute to an increase in blood pressure. Thus, polymorphic variants of the HSP70 family genes are associated with the risk of AH in patients with severe COVID-19.
GWAS-significant loci for severe COVID-19 affect biochemical parameters in patients with coronary artery disease
Severe COVID-19, including hospitalization and mortality, is determined by both genetic risk factors and concomitant chronic diseases, among which coronary artery disease (CAD) plays a leading role. The aim of this study was to evaluate the impact of CAD on the associations between biochemical markers of patients with severe COVID-19 and GWAS-significant loci. Genotyping of 199 patients hospitalized with severe COVID-19 was carried out by PCR for 10 GWAS loci. In the presence of concomitant CAD in patients with severe COVID-19, polymorphic variants were associated with an increase in plasma fibrinogen: rs12610495 DPP9 (β = 1.528±0.5637, Pperm = 0.02), rs7949972 ELF5 (β = 1.345±0.5015, Pperm = 0.02), rs61882275 ELF5 (β = 1.345±0.5015, Pperm = 0.02); and with an increase in the steadystate spatial clot growth rate: rs11183780 CCHCR1 (β = 14.36±6.5, Pperm = 0.03). Thus, CAD modifies associations between biochemical markers and GWAS-significant loci in patients with severe COVID-19.
The correction of SMN2 gene splicing using antisense oligonucleotides delivered with peptide carriers into SMA fibroblast cultures
Background. Spinal muscular atrophy is a severe neurodegenerative disease for which the development of therapeutic approaches and optimization of the delivery of therapeutic molecules into cells remains relevant. Aim: to investigate the effectiveness of correcting the splicing of the SMN2 gene with antisense oligonucleotides delivered in combination with a peptide carrier into fibroblast cultures of SMA patients. Methods. Fibroblast culture of SMA patient, antisense RNA oligonucleotides, peptide carrier. Methods of cell culture cultivation, RNA isolation and analysis, immunocytochemistry. Results. We have shown the effectiveness of antisense oligonucleotides (ASO) for correcting the splicing of the SMN2 gene delivered by a peptide carrier containing a ligand to receptors on the surface of target cells. A significant increase in the proportion of full-length SMN transcripts and gems containing the SMN protein was found as a result of the delivery of ASO:carrier complexes to SMA fibroblast cultures. Conclusion. These results are promising for the development of methods for the non-viral delivery of therapeutic molecules into the cells of SMA patients.
Long contiguous stretches of homozygosity detected by chromosomal microarrays (CMA) in patients with an abnormal phenotype and no clinically significant CNVs
We present data from ROH analysis in a cohort of patients without clinically significant CNVs. The study cohort consisted of 561 patients. Multiple ROHs exceeding 1% of the total autosomal genome were observed in 8 patients and in 8 cases single or multiple ROHs larger than 7Mb were localized on the same chromosome.
Haplotype frequencies of polymorphic variants of the NFKB1 gene in pulmonary tuberculosis
The NF-kB signaling pathway is one of their key mechanisms that trigger the synthesis of pro-inflammatory factors in response to infection with pathogens. The data on the study of haplotype frequencies of the NFKB1 gene by a panel of polymorphic variants are discussed: rs28362491, rs160993, rs4648050, rs4648051, rs4648055, rs4648058, rs4648068 in a sample of patients with pulmonary tuberculosis (TB, n=169) and in the population control group (PC, n=96). DNA isolated from blood samples using phenol-chloroform extraction, genotyped by real-time PCR. 49 haplotypic variants were identified in the group of patients, 33 in the control group. The spectrum and frequency of major haplotypes were determined: in the TB group – DCCGACG, ICTAGGA with a total frequency of 43%; in the PC sample – DCCGACG, ICTAGGA, ICTAGCA and DCCGAGG (a total frequency of 60%).
Genetic disorders in the etiology of CAKUT. Multicenter study (Moscow-Minsk)
Congenital anomalies of the kidney and urinary tract (CAKUT) account for an average of 25% of all genetic defects diagnosed in utero. The incidence of CAKUT is 2–6 cases per 1000 newborns. To form a prognosis for the disease and determine the tactics of introducing patients into clinical practice, as well as medical and genetic counseling, it is necessary to diagnose monogenic forms of pathology using medical exemptions of high-throughput next-generation sequencing, which has demonstrated high efficiency in identifying pathogenic mutations associated with CAKUT.
46,XY disorders of sex development in adolescents: V.I. Kulakov Center experience
Disorders of sex development (DSD) are a genetically and phenotypically heterogeneous group of developmental disorders characterized by phenotypic and chromosomal sex mismatch. In the period from 2022-2024, 34 adolescent female patients with DSD with karyotype 46,XY were admitted to the 2nd Gynecological Department of the Kulakov Center. The patients were analyzed for the presence of the SRY gene and, if positive, exome sequencing was performed to determine the monogenic cause of the disease. The genetic cause of DSD was determined in 26 of 34 patients (76% effectiveness).
Experience with canakinumab in patients with familial mediterranean fever
Familial Mediterranean Fever (FMF) is the most common monogenic autoinflammatory disease, associated with mutations in the MEFV gene, characterized by recurrent febrile attacks, abdominal pain, arthritis, and systemic inflammation. The high risk of amyloidosis necessitates pathogenetic therapy, especially in colchicine-resistant cases. This report presents clinical experience with the use of canakinumab in three children with colchicine-resistant FMF. Two patients, including a 3-year-old girl, showed significant clinical and laboratory improvement after 4 and 12 weeks of therapy. No serious adverse effects were observed during follow-up. The treatment involved subcutaneous injections of 2 mg/kg of canakinumab once every 4 weeks, resulting in remission and reduced systemic inflammation. These findings confirm the safety and efficacy of canakinumab in pediatric patients with severe colchicine-resistant FMF, expanding therapeutic options for this disease.
Epidemiological, clinical and genetic characteristics of patients with hereditary motor and sensory neuropathy 1X according to regional registry data
The article presents updated data on the epidemiology, clinical and genetic characteristics of patients with hereditary motor-sensory neuropathy 1X, obtained as a result of continued work with the regional genetic registry.
Modern possibilities of рroviding medical-genetic care in a family with rare hereditary syndrome EEC: a case report
The article presents a description of a clinical case of a rare hereditary syndrome «Ectrodactyly-ectodermal dysplasia with cleft lip/ palate» (EEC3, OMIM #604292). Modern capabilities of NGS sequencing for identifying pathogenic variants of nucleotide sequences in the TP63 gene are demonstrated. A pathogenic variant was identified in the proband, which confirmed the diagnosis and allowed the family to plan childbearing.
Study of the effect of spermidine on the genotoxicity of dioxidine and hydrogen peroxide using sensory strains of Escherichia coli
The effect of spermidine on the DNA-damaging effect of dioxidine and hydrogen peroxide was studied on the cells of the lux-biosensors E. coli MG1655 (pColD-lux) and (pKatG-lux), luminescent as a result of activation of the promoters of the colicin colD and catalase katG genes. It was shown that spermidine reduces the genotoxic effect induced by the studied drugs by 40 and 23% in E. coli cells, and also reduces the number of DNA breaks.
Study on the relationship between glutathione synthetase gene polymorphisms and the risk of coronary artery disease
The aim of this study was to investigate the relationship between three functionally significant single nucleotide polymorphisms (SNPs) rs7265992, rs6088660, and rs1801310 of the GSS gene and the risk of coronary heart disease (CHD). The findings indicated that the rs7265992 and rs6088660 polymorphisms were associated with a reduced risk of CHD, whereas the rs1801310 variant was associated with an increased risk of developing the disease. Stratified analysis by gender revealed that the rs1801310 polymorphism of the GSS gene was linked to an increased risk of CHD only in men. This study establishes for the first time that polymorphic variants of the glutathione synthetase gene are associated with a predisposition to coronary heart disease.
Establishment of a mouse model of mucopolysaccharidosis-plus syndrome using CRISPR-Cas9 technology
Mucopolysaccharidosis-plus syndrome (MPSPS) is an autosomal recessive disease caused by a missense variant in the VPS33A gene. The aim of this work is to create a mouse model of MPSPS by introducing the target variant using CRISPR-Cas9 technology. Sperm and oocyte donors were mice of the C57BL/6J strain. The CRISPR-Cas9 system was introduced by electroporation. Litter birth was observed on days 19-21. Validation of successful variant introduction was performed by PCR-RFLP and Sanger sequencing. This resulted in one mouse (male) in which the p.R500W variant in the Vps33a gene was introduced into the genome. After several crosses, heterozygous and homozygous mice were obtained. Homozygous mice were developmentally retarded. This may be due to the influence of the variant on intrauterine development.
Turner syndrome caused by a pseudo-isodicentric Xp chromosome containing two copies of the SHOX gene
The article describes a clinical case of Turner syndrome (TS) in a 15-year-old patient with mosaicism involving a pseudoisodicentric Xp chromosome. The disorder was characterized by delayed puberty, primary amenorrhea, gonadal dysgenesis, uterine hypoplasia, hypergonadotropic hypogonadism, and stigmas of dysembryogenesis. Comprehensive cytogenetic, molecular-cytogenetic, and molecular-genetic studies were conducted. Mosaicism with two cell lines was found in lymphocytes and buccal epithelial cells: one with X chromosome monosomy, the other with one normal X and one pseudoisodicentric Xp chromosome carrying two copies of the SHOX gene. Chromosomal microarray analysis detected monosomy of most of the Xq, and trisomy of the Xp and proximal region of the Xq, with a breakpoint at the Xq13.3 locus (75,444,376–155,233,731). The patient’s normal growth is attributed to the presence of two copies of the SHOX gene on the derivative chromosome. This clinical case enhances the understanding of the genetic mechanisms underlying the phenotypic manifestations of patients with pseudoisodicentric Xp chromosomes.
Identification of the classic type of Ehlers-Danlos syndrome
Background. Ehlers–Danlos syndrome (EDS) is an inherited connective tissue disorder most commonly associated with mutations in type V collagen genes. However, data on the spectrum of hereditary variants among patients from Russia remain fragmentary. Aim: to identify mutations in type V collagen genes in patients with Ehlers–Danlos syndrome from the Republic of Bashkortostan. Methods. DNA samples from 43 patients (mean age 28.6 ± 7.44 years) representing 37 families with Ehlers–Danlos syndrome residing in the Republic of Bashkortostan were analyzed. Results. Five nucleotide sequence alterations were detected in the COL5A1 gene and three in the COL5A2 gene. Pathogenic variants c.212delC (p.Pro71ArgfsTer33) and c.4135C>T (p.Pro1379Ser) in COL5A1 were identified, which have not been previously described in the literature. Conclusions. Mutations were identified in eight unrelated families with EDS from the Republic of Bashkortostan. In total, five nucleotide sequence alterations were found in the COL5A1 gene and three in the COL5A2 gene.
CNV load in families with a child with a neurodevelopmental disorder and a history of miscarriage
Copy number variation (CNV) found in patients with neurodevelopmental disorders (NDD) are also detected in miscarriages. The study was performed for CNVs associated with both the birth of a child with NDD and spontaneous abortion (SA). CNVs have been shown to be more common in patients born to mothers with a history of SA (p = 0.03). Microduplications at 2q23.1, 3q29, 16p11.2 are likely to affect both prenatal and postnatal development.
Diagnostics of sperm head morphology genetic abnormalities in primary male infertility
Male infertility is often associated with impaired spermatogenesis that leads to a decline in sperm quality and quantity. According to the WHO laboratory manual for the examination and processing of human semen, published in 2021, teratozoospermia is diagnosed if 4% or fewer sperm display normal shapes. Monomorphic teratozoospermia is a group of rare genetic sperm anomalies, leading to male infertility. The results of comprehensive semen and genetic examination of four infertile men with globozoospermia (n=3) and acephalic sperm syndrome (n=1) are presented. WES revealed a homozygous likely pathogenic nonsense DPY19L2 gene mutation (chr12:63976191G>A) and homozygous missense-variants of uncertain significance in the DPY19L2 (chr12:64038264G>A) and the SPATA16 (chr3:172766742 T>C) genes in globozoospermic patients. A patient with acephalic sperm syndrome was found to have two likely pathogenic compound heterozygous variants (chr16:72123598G>A and chr16:72130275G>A) in the PMFBP1 gene.
Application of bacterial test systems and C. elegans nematodes for studying the genotoxicity of the photosensitizer 8-methoxypsoralen
The lux-test method showed that the combined effect of 8-methoxypsoralen (8-MOP) and UV irradiation leads to an increase in the luminescence of lux-biosensors by 3-40 times compared to the control. The nematode test confirmed the hypothesis of induction of active oxygen species (AOS) in eukaryotes when exposed to 8-MOP in combination with UV irradiation.
Monogenic causes of premature ovarian insufficiency in adolescence
Premature ovarian insufficiency (POI) is a condition characterized by hypergonadotropic hypogonadism and amenorrhea, which rarely manifests in adolescence. Up to 50% of POI cases are idiopathic. We conducted molecular diagnostic testing of 39 patients who experienced the onset of POI before the age of 18, all of whom had a normal female karyotype. This involved FMR1 CGG repeats testing and whole-exome sequencing. None of the patients were found to have a premutation in the FMR1 gene. In 17 out of 39 patients (43.5%), variants were identified across 12 genes. A diagnosis of monogenic POI was established in 7 patients (18%) based on the detection of pathogenic and likely pathogenic variants. Syndromic forms of POI were observed in 3 cases (8%), while isolated POI was noted in 4 cases (10%). In 4 patients (10%), variants of uncertain clinical significance were found in the STAG3, LMNA, SPIDR, and NOBOX genes. Additionally, 6 patients (15.5%) had heterozygous variants on one allele in genes associated with autosomal recessive POI. We recommend that patients with onset of POI in adolescence undergo genetic counseling and molecular genetic testing.
Prenatal diagnosis of hereditary pathology: to terminate or to prolong pregnancy
Prenatal diagnosis using molecular genetic technologies is crucial for early detection of genetic abnormalities but faces medical and ethical challenges, including limited information on hereditary diseases and therapeutic options, forcing families to choose between terminating or continuing pregnancy. This study presents a case of non-immune hydrops fetalis (NIHF) diagnosed at 27 weeks. Amnioreduction at 28 weeks and 5 days was followed by NGS, revealing a PTPN11 variant linked to RASopathies (Noonan and LEOPARD syndromes). Placental abruption at 31 weeks led to perinatal death. The patient declined further testing, citing a healthy child. The case highlights the emotional and diagnostic complexities of prenatal care, emphasizing the need for improved patient education and expanded therapeutic options.
Obesity modifies the effects of SERBP1 polymorphisms on hemocoagulation and coronary artery stenosis
In the pathogenesis of coronary heart disease (CHD), which develops against the background of progression of atherosclerosis, the participation of molecular chaperones is one of the determining factors. SERBP1 (Hero45), an RNA-binding protein, has recently been classified as a new class of Hero proteins with chaperone-like activity, which makes its study particularly relevant. In the present study, 836 patients with CHD were genotyped for five single nucleotide polymorphisms of SERBP1 using PCR. In this study, we report that single nucleotide polymorphisms of the SERBP1 gene in non-obese patients (BMI < 30) are associated with an increase in international normalized ratio (INR) (rs4655707 SERBP1: β = 0.4191 ± 0.1661, Pperm = 0.007; rs6702742 SERBP1: β = 0.4195 ± 0.1719, Pperm = 0.0098) and a decrease in the percentage of left coronary artery (LCA) stenosis (rs12561767 SERBP1: β = -1.277 ± 0.4637, Pperm = 0.02; rs6702742 SERBP1: β = -1.277 ± 0.4637, Pperm = 0.02). Taken together, our study provides evidence that SERBP1 may modulate hemostasis parameters and stenosis in non-obese CAD patients.
Polymorphism rs1136141 HSPA8 is associated with an increase in the brain infarct size
Ischemic stroke (IS) is one of the leading causes of disability and mortality. A key role in its pathogenesis is played by impaired proteostasis and, accordingly, impaired functions of heat shock proteins (HSP). In this study, we sought to evaluate the impact of polymorphisms of the HSPA8 gene encoding heat shock protein on the clinical course of IS. DNA samples of 888 patients with IS were genotyped by real-time PCR at three loci of the HSPA8 gene (rs1461496, rs10892958, rs1136141). The analysis showed that the rs1136141 HSPA8 polymorphism is associated with an increase in the brain infarct size (β = 112.5±54.07, Pperm = 0.04). Bioinformatic analysis of this polymorphism established that the A allele of rs1136141 HSPA8 may be associated with the regulation of the immune response, apoptosis and angiogenesis, which in turn may contribute to tissue damage during ischemia/reperfusion.