One of the main problems in the development of gene therapy methods based on adenovirus constructs is the immune response to the introduction of viral particles. It is proposed to use the non-steroidal anti-inflammatory drug ibuprofen to reduce the immune response. Ibuprofen at high concentrations has a dose-dependent toxic effect on cultures of multipotent mesenchymal stromal cells (MSCs). It was determined that at concentration of 0,125 mg/ml ibuprofen does not cause cell death, and also has a positive effect on adenovirus transduction of MSCs: 48,47±2,06% of transduced cells were detected 3 days after infection with 320 TCID50/ml Ad-GFP, and when 0,125 mg/ml Ibu was added - 60,07±1,64% (p < 0,05). Ibuprofen can be used in gene therapy based on adenoviral transduction to reduce the immune response to local exposure high doses of viral vectors.

Previously, we identified a novel disease from the group of hereditary metabolic diseases with an autosomal-recessive type of inheritance - mucopolysaccharidosis-plus syndrome (MPSPS). Using whole exome sequencing, we revealed pathogenic homozygous mutation p.R498W in the VPS33A gene. The objective of study was to analyze the effect of the mutation in the VPS33A gene on the protein level and its function. The cellular model of the disease was generated using Crispr-Cas9 system. Determination of autophagic activity was carried out by analyzing differences in the amount of the LC3-II between samples with and without addition of lysosome inhibitor bafilomycin A1. The level of endocytosis was determined by analysis of EGFR degradation. In the present study we revealed a reduction in the level of the VPS33A protein in cell lines with heterozygous and homozygous p.R498W mutations up to ~43% and ~25%, respectively. We also found that p.R498W mutation does not affect to the main known functions of the VPS33A - endocytosis and autophagy. The fluorescence intensity in the mutant cells was significantly increased compared to the controls, which indicated a shift of lysosomal pH to the acid side. Impairment of domain 2-2 led to dysfunction of the VPS33A protein. The fundamental significance of the study is due to decipher the pathological mechanisms of disease development to find approaches to adequate therapy.

Purpose of the study: to determine the impact of rDNA fragments in the culture medium on the rate of replicative cell aging. We found that GC-rich fragments in extracellular DNA accelerate the replicative aging of cultured human skin fibroblasts and increase the level of reactive oxygen species in cells.

Glucocerebrosidase (GCase) is a lysosomal enzyme encoded by the GBA gene, mutations in which are the most common genetic risk factor for Parkinson’s disease (PD). Homozygous carriage of mutations in the GBA gene leads to the development of Gaucher disease (GD). It has been shown that mutations in the GBA gene can affect the accumulation of alpha-synuclein protein. Its aggregation is currently considered as a key link in the pathogenesis of PD. In this study, we compared the enzymatic activity of GCase, the concentration of the HexSph enzyme substrate and the level of alpha-synuclein protein in the primary culture of macrophages of patients with GD, GBA-associated PD, asymptomatic carriers of mutation in the GBA gene and control group individuals in the presence of a selective GCase inhibitor conduritol-B-epoxide, as well as in the brain cells of model animals with lysosome dysfunction. As a result of the study conducted on the primary culture of macrophages, as well as on animal models, it was shown that lysosome dysfunction leads to decreased GCase activity and HexSph accumulation. In this study on model animals, we have shown for the first time the effect of GCase function inhibition on the oligomeric forms of alpha-synuclein accumulation.

It was previously found that in the blood leukocytes of patients with schizophrenia, compared to the control, the content of the tandem repeat - the subfraction of satellite III (SatIII), which is part of the pericentromeric heterochromatin of the first chromosome (region 1q12) - is significantly reduced. One of the reasons behind the low repeat level in the DNA of patients may be the low level of SatIII transcription under stress conditions. The study shows that cellular stress in cultured skin fibroblasts of patients with schizophrenia and healthy people, caused by an increase in cultivation temperature, significantly blocks the transcription of SatIII (1q12) and increases the transcription level of SatIII on the 9th chromosome tenfold. For cells of patients with schizophrenia, a decrease in the content of SatIII(1q12) in DNA positively correlates with a decrease in the amount of SatIII RNA. In control cells, the content of SatIII in DNA did not change under stress. Blocking of SatIII transcription under stress conditions can potentially be one of the reasons for the decrease in the content of the SatIII repeat in the DNA of patients with schizophrenia.

To date, there are no neuroprotective drugs for the common neurodegenerative disease, Parkinson’s disease (PD). PD associated with mutations in the GBA gene (GBA-PD) is the most common form of PD with a known etiology. GBA-PD is considered the most promising for the development of therapy for PD. Mutations in the GBA gene encoding the enzyme glucocerebrosidase (GCase) lead to a decrease in the activity of this enzyme. Previously, it was shown that inhibition of LRRK2 kinase activity by MLi-2 inhibitor leads to an increase in GCase activity. In this study, we showed for the first time the effect of the LRRK2 kinase activity inhibitor MLi-2 not only on the activity of GCase, but also on the activity of other lysosomal enzymes in the primary culture of peripheral blood macrophages of patients with LRRK2-associated PD (LRRK2-PD) and GBA-PD.

Age-related macular degeneration (AMD) is a complex neurodegenerative disease that becomes the main cause of central vision loss in people over 60 years of age. The development of AMD is controlled by a variety of interacting genetic and environmental factors that determine the form and rate of progression of the disease. Epigenetic mechanisms also significantly affect these parameters, including changes in microRNA expression patterns, the assessment of the circulating pool of which is considered as a promising approach to early diagnosis, prognosis of the course and evaluation of the effectiveness of AMD treatment. However, there is practically no information about the patterns of microRNA expression in the retina at different stages of AMD development, especially preclinical ones, and it can only be obtained using adequate biological models. The present study was performed on a line of OXYS rats (ICG SB RAS) - a unique model of premature aging, one of the manifestations of which is the development of a complex of key signs of AMD. Its purpose was to evaluate changes in the expression of a number of prognostically promising microRNAs (miR-9, miR-27a, miR-34a, miR-146a, miR-155) with age in the retina of Wistar rats (control) and OXYS with the development of AMD-like retinopathy. microRNA expression was evaluated by real-time PCR An increase in the level of miR-27a expression in the retina of OXYS rats was revealed with age and with the development of signs of retinopathy, which may contribute to the development of signs of AMD.

The gene pool of Tomsk Tatars living in three settlements was studied: the village of Chernaya Rechka, the village of Takhtamyshevo, and the village of Eushta. It was revealed that the Tomsk Tatars are the most heterogeneous group in Siberia. The obtained results of the gene pool of the Tomsk Tatars reflect the complex process of their formation on the basis of ancestral population groups of different origins. According to the results of the analysis of genetic differentiation according to the frequencies of haplogroups, when dividing the Tomsk Tatars into three samples in accordance with the settlements, a high degree of their intergroup differences was shown. Tomsk Tatars differ significantly from other groups of Siberian Tatars in terms of the composition of haplogroups. According to different haplogroups, the connection of Tomsk Tatars with Teleuts, Northern Altaians, Shors, Khakass, Tuvans and Buryats is shown, which confirms their connection with the South Siberian Turkic-speaking and Mongolian-speaking peoples. A significant proportion of the gene pool inherited from people from the Volga region and Bukharians from Central Asia was also revealed.

Based on the analysis of mitochondrial DNA sequences deposited in databases, the spectrum of amino acid variability of the mtDNA-encoded subunits of the respiratory chain complex I was studied depending on mtDNA haplogroup affiliations. It was found that about 20% of amino acids were polymorphic in the studied dataset, and for 30% of polymorphic amino acids more than one mutational event (homoplasy) was registered. The effect of negative selection was revealed for some complex I genes in mtDNA haplogroups, distributed mainly in Northern Eurasia. It has been shown that the conservation index for amino acid substitutions in complex I genes is lower than in other mtDNA genes, and for the ND1 gene this index is higher than for other complex I genes. For the MutPred index and the mtDNA selection score, which reflect the effect of amino acid substitution on the structure and function of the protein, higher average values were registered in the “Northern” mtDNA haplogroups, comparing to the “Southern” haplogroups.

The results of a study of the population frequencies of polymorphic variants of the VDR gene (FokI, rs2228570; BsmI, rs1544410) in populations of Tobolo-Irtysh Siberian Tatars and Shors are discussed. The material for the study was collected in the expeditions of the scientific groups of KemSMU of the Ministry of Health of the Russian Federation, MGNC, KemSU, Tobolsk Integrated Scientific Station of the UrB of RAS. The total sample size is 166 people. Genotyping was carried out by real-time PCR using TaqMan probes. Mathematical processing - using the STATISTICA 8.0 statistical software package and standard approaches of population genetics. The study showed that the population of Shorets (the area of settlement in the Tashtagolsky district of the Kemerovo region) they are characterized by lower frequencies of minor alleles of the VDR BsmI*A and FokI*C in comparison with the Tobolo-Irtysh Siberian Tatars (Tyumen, Yarkovsky, Yalutorovsky districts of the Tyumen region). In the studied groups, there is a deviation in the level of heterozygosity.

The fund of surnames of Iskero-Tobolsk, Istiak-Tokuzsk, Sargatsko-Utuz Tobolo-Irtysh Tatars and Siberian Bukharans for the 1950s, 1980s and 2010s was studied (18630 people, over 1150 surname variants). The surveyed groups territories of settlement are located in contact zones - the groups are geographically connected to each other and are characterized by “reciprocal” migrations, which can cause the “erasure” of the clear boundaries in population gene pools. Surnames, as an analogue of genetic markers, are used to study the transformation of the population-genetic structure of Siberian-Tatar population in time and geographical space. The acquired results testify to the distinctive identity of surname composition of the ethnographic groups studied and confirm the effectiveness of Tatar surnames as a tool in assessing the transformation of their populations structure: not only to record events, but also to understand their causes. The data analysis of surname funds based on genetic distances is consistent with the geography of their ethnic habitats within the contact zones and characterize the prospects of using surnames to solve problems of population genetics.

Loss of ACTN3 gene activity due to a stop codon at the rs1815739 locus (rs1815739-T or 577X allele) leads to the absence of α-actinin-3 in fast-twitch muscle fibers, which contributes to increased endurance and resistance to cold. These circumstances became the basis for the hypothesis that the increase in the frequency of the rs1815739-T allele in the populations of Eurasia and America, in comparison with Africa, occurred under the influence of selection at the initial stages of human exploration of the planet. In this work, we analyzed the polymorphism of the rs1815739 locus in the populations of Siberia - in the south (Buryats, Altaians, Tuvinians, Todjins) and north (Chukchi, Koryaks, Evens, Evenks) of the region. We have shown that, contrary to the above hypothesis, the frequency of the rs1815739-T allele is higher in the south of Siberia, while in the northern direction, on the contrary, the frequency of the rs1815739-C allele and the CC genotype increases. The results of the study suggest that the distribution pattern of the rs1815739 polymorphic variants in Siberian populations is not associated with adaptation to cold.

The contribution of the Permian by origin component to the gene pool of various ethnic groups of the Volga region, which is present among the peoples of the Permian language group and Turkic-speaking ethnic groups, is revealed. It completely dominates on Udmurts and Besermyans, and also occupies a fifth of the Komi gene pool. An analysis of the distribution of this component in populations and the composition of their gene pools for various sublines of the Y-chromosome haplogroups shows that modern Udmurts retained their original composition of the gene pool and were not subjected to mixing with the Ugric, Turkic and Slavic settlers who came later. The introduction of this genetic component into the composition of the Tatars and Bashkirs was associated with the assimilation of the local Permian population. The distribution of frequencies of the Permian component in populations according to genomic data and the frequencies of haplogroups N1a1a and N1a2b in the populations of the Udmurts, Besermians and Komi completely coincides with the data of anthropology and linguistics.

Intragenic deletions and duplications are a major problem in the interpretation of the chromosomal microarray results in patients with suspected monogenic disease. Assessing the clinical significance and causation of such genetic variants, especially extended duplications, is difficult because CNV`s affecting one or more gene exons can have different phenotypic effects. The article presents the results of the examination of two patients with an incoming diagnosis of muscular dystrophy. Intragenic duplications in the DMD gene were diagnosed in both patients during chromosomal microarray analysis. The paper discusses the interpretation features of such CNVs and suggests recommendations for confirmation of the of muscular dystrophy genetics diagnosis.

The 8p23.1 paracentric inversion is the largest polymorphic inversion in the human genome. As a result of meiotic malsegregation of paracentric inversion, gametes and subsequently zygotes are formed with recombinant chromosomes, namely with a chromosomal genomic imbalance such as an inverted duplication with an adjacent deletion (inv dup del) or with a terminal deletion (del). In this study, for the first time, the frequency of recombination in the inversion loop was estimated in a carrier of polymorphic paracentric inversion 8p23.1. It was shown that the frequency with which meiotic recombination happens in male gametogenesis is 0.03%.

Introduction. Small supernumerary marker chromosomes (sSMCs) consisting of heterochromatin regions can be found in individuals without visible phenotypic abnormalities. Their presence in the karyotype sometimes leads to reproductive problems associated with the risk of meiosis errors due to interchromosomal synapses and the process of non-disjunction of homologous chromosomes. Aim: to investigate meiotic segregation and evaluate the frequency of gamete formation with aneuploidy in males with karyotype 47,XY,+mar without clinically significant phenotypic abnormalities. Materials and Methods: fluorescence in situ hybridization (FISH) on fixed sperm preparations in 7 male patients-asymptomatic carriers of sSMC. Results. In all sSMC carriers, the frequency of spermatozoa containing the marker chromosome did not exceed 30% and averaged 15.04±9.4 (±SD). The proportion of aneuploid spermatozoa did not exceed 3.1% and, on average, was 1.9%±1.1, which does not differ from the frequency of aneuploidy in gametes in individuals with normal karyotype (does not exceed 5.9% and, on average, is 2.2±1.8). Conclusions. In carriers of heterochromatinous sSMC gametes in meiosis are predominantly formed without a marker chromosome. It has been shown that the presence of a high proportion of sSMC in the ejaculate cells does not significantly affect the meiotic segregation of other chromosomes, i.e., there is no interchromosomal effect.