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Objective : to elucidate whether cerebrolysin contains peptide fragments that promote geroprotection. Material and methods. The peptide composition of cerebrolysin underwent a comprehensive mass spectrometric analysis, followed by a systemic biological assessment. Results and discussion. Thirty-six peptides with geroprotective properties were isolated in the multipeptide composition of cerebrolysin. These peptides included those of mimetics of adrenomedullin and enkephalins; those of inhibitors of seven targeted human proteins (the protein kinases MAPK1, VPRBP, and PKC, the gamma-secretase PS1, the kinases CDK1, SGK1 and mTOR). The established cerebrolysin peptides were shown to be competitive inhibitors of the seven targeted proteins. In particular, inhibition of PKC and mTOR stimulated the increased autophagy (the utilization of waste and abnormal proteins), which contributes to an increase in the survival of cells and model organisms. Conclusion . Cerebrolysin can have geroprotective effects, by inhibiting the seven targeted proteins, by activating endorphinergic neurotransmission, and can be indirectly involved in vascular tone normalization.

Low grade gliomas are the most common brain tumors in children. Total resection for operable lesion helps to achieve local and system control. Nevertheless, for inaccessible tumors are required more effective treatment both to overcome the refractory course of the disease, and to mi nimize toxicity with conventional adjuvant chemotherapy and various types of radiation therapy. In recent years, there has been an accelerated understanding of the molecular pathogenesis of some tumors in children, including low grade gliomas. Given the fact that the basis of the molecular pathogenesis of the low grade gliomas is the activation of signaling pathways MARK (mitogen activated protein kinase) and mTOR (mammalian target of rapamycin), the most promising targeted agents are BRAF, MEK and mTOR inhibitors. Nevertheless, a number of other agents have been studied to find promising targeted therapy for this tumors type. This article summarizes the latest literature evaluating new drugs in low grade glioma.

Introduction. Cutaneous melanoma is a challenge to treat due to rapid progression of disease and acquired resistance to therapy. Autophagy and the epithelial-to-mesenchymal transition (EMT) are closely interrelated and play a key role in tumor progression. Targeted co-inhibition of MEK and mTOR kinases is a potential target for melanoma therapy by downregulatoin of the EMT.Objective: to study the effect of MEK and mTOR co-inhibition on cell viability, ability to form 3D-spheroids and migratory capacity of melanoma cell lines, and correlation of these changes with EMTand autophagy-related markers.Material and Methods. Melanoma cell lines Mel Z and Mel MTP were derived from patients, who were treated at the N.N. Blokhin National Medical Research Center of Oncology. The antiproliferative effect of binimetinib and/or rapamycin was studied by the MTT -test. 3D spheroids were formed using RGD peptides. Cell migration and invasion were assessed by a Boyden chamber migration assay. The expression levels of autophagy and EMT markers were investigated by immunocytochemistry or immunoblotting.Results. Rapamycin increased cytotoxicity of binimetinib in both 2D and 3D melanoma cell line cultures. At the same time, binimetinib and rapamycin reduced invasion, but not migration capacity of melanoma cells in vitro. The effectiveness of the combination was associated with a decrease in the EMT markers (N-cadherin and β-catenin) and autophagy markers (Beclin 1, p62/SQST M1 and LC3BII ) in melanoma cells.Conclusion. Inactivation of autophagy and EMT leads to overcoming the resistance to current anti-melanoma therapy and can be considered as a promising target for the treatment of melanoma.

The modern concept of therapy for intrahepatic cholangiocarcinoma including surgical treatment, must take into account the achievements of molecular biology and modern staging principles. A detailed understanding of the molecular genetic (genetic and epigenetic) disorders underlying the pathogenesis of cholangiocarcinoma is important, which will improve the results of surgical treatment and expand the possibilities of personalized (targeted) therapy. Based on new data on cholangiocarcinogenesis, molecular profiling of bile duct tumors may be most appropriate for the selection of treatment in cases refractory to standard therapy. Current potential target therapy targets include endothelial growth factor receptors, fibroblast growth factor, MET tyrosine kinase, PI3K/Akt/mTOR signaling pathway and isocitrate dehydrogenase mutation. The review considers the molecular-genetic aspects underlying the pathogenesis and modern principles of staging intrahepatic cholangiocarcinoma.

Many patients with osteoarthritis (OA) tend to have comorbidities. This tendency is more frequently observed to increase with age. One of the comorbidities is type 2 diabetes mellitus (T2DM). Due to the higher prevalence of coexistence of these two conditions, it has been suggested that T2DM-associated hyperglycemia may adversely affect joint tissues and increase OA severity. However, the molecular mechanisms in the development of DM in patients with OA remain unclear. Objective : to trace the dynamics of T2DM development in patients with OA at the level of the expression of genes associated with glucose metabolism, joint destruction, and general regulation of metabolic processes. Patients and methods . Three patients with OA were followed up for 4—6 years, including the year of onset of T2DM. The clinical condition of the patients was analyzed once a year. Total RNA was annually isolated from their blood and used to determine the level of gene expression by real-time polymerase chain reaction. Results and discussion. The development of T2DM was shown to be accompanied by the increased expression of genes related to glycolysis, Krebs cycle, pentose phosphate pathway, matrix metalloproteinases and regulators of AMPKand mTOR metabolism. By contrast, the level of the hypoxia regulator HIF1a and hexosamine pathway genes was decreased. Conclusion . The occurrence of T2DM in the presence of OA is likely to be associated with the higher needs for cells for ATP energy and is accompanied by activation of the glucose assimilation pathways, as well as by the increased expression of the genes responsible for extracellular matrix destruction. This may be caused by impaired protein glycosylation due to inhibition of the hexosamine pathway.

The exosomes involvement in the pathogenesis of tumors is based on their property to incorporate into the recipient cells resulting in the both genomic and epigenomic changes. Earlier we have shown that exosomes from different types of estrogen-independent breast cancer cells (MCF-7/T developed by long-term tamoxifen treatment, and MCF-7/M) developed by metformin treatment were able to transfer resistance to the parent MCF-7 cells. To elucidate the common features of the both types of resistant exosomes, the proteome and microRNA cargo of the control and both types of the resistant exosomes were analyzed. Totally, more than 400 proteins were identified in the exosome samples. Of these proteins, only two proteins, DMBT1 (Deleted in Malignant Brain Tumors 1) and THBS1 (Thrombospondin-1), were commonly expressed in the both resistant exosomes (less than 5% from total DEPs) demonstrating the unique protein composition of each type of the resistant exosomes. The comparative analysis of the miRNA differentially expressed in the both MCF-7/T and MCF-7/M resistant exosomes revealed 180 up-regulated and 202 down-regulated miRNAs. Among them, 4 up-regulated and 8 down-regulated miRNAs were associated with progression of hormonal resistance of breast tumors. The bioinformatical analysis of 4 up-regulated exosomal miRNAs revealed 2 miRNAs, mir- 101and mir-181b, which up-regulated PI3K signaling supporting the key role of PI3K/Akt in the development of the resistant phenotype of breast cancer cells.

Альтернативные сигнальные механизмы активации транскрипционного фактора HIF-1 принципиально можно подразделить на 3 типа: связанные с ингибированием пролилгидроксилаз и ограничением деградации HIF-1 связанные с индукцией HIF-1 связанные с активацией HIF-1. Особый практический интерес представляет активация фактора HIF-1 во время гиперкапнии, которая часто сопутствует гипоксии. Пермиссивная гиперкапния вызывает повышение концентрации HIF-1 в ткани головного мозга и потенцирует активацию HIF-1, что выражается в повышении содержания HIF-1 в гиппокампальных нейронах СА1 региона после гиперкапнически-гипоксического воздействия. При изолированном применении гипоксии аналогичный эффект отсутствует. Перспективным для разработки методов повышения толерантности органов и тканей к гипоксии/ишемии является комбинирование средств, повышающих активность HIF-1, с респираторными гиперкапнически-гипоксическими тренировками и препаратами, влияющими на другие адаптогенные механизмы., Along with the classical oxygen-dependent mechanism of activation of the transcription factor HIF-1, there are alternative signaling mechanisms of this process. These mechanisms can be fundamentally divided into the following types: associated with the inhibition of prolyl hydroxylases and limiting the degradation of HIF-1 (iron chelators, NO, succinate, inhibitors of lysosomal degradation) associated with the induction of HIF-1 (TNF, IL-1, angiotensin) associated with the activation of HIF-1 (MAPK, PI3K). Of particular practical interest is the activation of HIF-1 factor when exposed to CO2 during hypercapnia, which is often associated with hypoxia. Permissive hypercapnia causes an increase in the concentration of HIF-1 in brain tissue at an equivalent level of hypoxia, and also potentiates the effectiveness of hypoxia in relation to HIF-1 activation, which is expressed in an increase in the content of HIF-1 in hippocampal neurons of the CA1 region after hypercapnic-hypoxic exposure, on the absence of a similar effect with the isolated use of hypoxia. Promising for the development of methods to increase the tolerance of organs and tissues to hypoxia / ischemia is a combination of agents that increase the activity of HIF-1, with respiratory hypercapnic-hypoxic training and drugs that affect other adaptogenic mechanisms (A1-adenosine receptors, mitoKATP-channels, opioid receptors)., №4 (2019)

Prostate cancer (PC) is one of the most common causes of death from malignant neoplasms in men in many countries around the world. Transmission of the signal in the androgenic axis of regulation is crucial for the development and progression of PC. Despite the constant dependence on androgen receptor signals in castration resistance, the use of new anti-androgenic drugs invariably leads to the stability of the ongoing treatment. The interaction of androgen receptor and alternative (phosphoinositide-3-kinases, PI3K) pathways in the regulation of cells can be one of the mechanisms of resistance to treatment. In this article, we describe current treatments for metastatic castration-resistant PC and the possible role of the PI3K pathway in the pathogenesis and progression of PC.

The last decade was marked by the rapid development of kidney cancer drug treatment and advent of targeted drugs aimed at inhibition of angiogenesis which plays a crucial role in tumor growth. Despite certain success, targeted antiangiogenetic therapy with tyrosine kinase inhibitors, mammalian target of rapamycin inhibitors (mTOR), and monoclonal antibodies against vascular endothelial growth factor (VEGF) in most cases do not achieve long-term remission, are highly toxic, and never lead to full cure for the patients. Development of modern immunological approaches to application of inhibitors of the crucial immune response regulators opens up new possibilities in treatment of disseminated kidney cancer. In this review, results of the studies of nivolumab (PD-1 inhibitor), first checkpoint inhibitor registered for treatment of metastatic kidney cancer are presented.

This article reviews recent advance in our understanding of ovarian endometriomas, with special reference of atypical epithelial features (atypical endometriosis) and its relationship to neoplasm. Mainly, two epithelial ovarian carcinoma subtypes, the ovarian clear cell carcinomas and the endometrioid ovarian carcinomas, have been molecularly and epidemiologically linked to endometriosis. The clear cell and endometrioid adenocarcinomas have common immunohistochemical markers, genetic changes are the same as atypical endometriosis. The occurrence of ARID1A mutations and alterations in the PI3K/AKT pathway in endometriosis and endometriosis-associated ovarian carcinomas, as well as the possible functional and clinical implications are discussed in this review. Therefore, development of a screening method that can detect mutations of ARID1A and activation of the PI3K/AKT pathway might enable early diagnosis of endometriosis associated ovarian cancers and endometrial cancers.

Osteoarthritis (OA) is a chronic disease associated with pain, stiffness, limited mobility and joint inflammation, as well as articular cartilage destruction. Recent studies have shown the importance of chondrocyte differentiation (hypertrophy) as one of the mechanisms of cartilage degradation in OA. This suggests that chondrocyte metabolism undergoes the profound changes during cartilage resorption, which are due to dysregulation of cell function. One of the major cellular metabolic regulators is the protein mTOR (mechanistic target of rapamycin) that controls cell growth, proliferation, protein biosynthesis and integrates extracellular signals from growth factors and hormones with amino acid availability and intracellular energy status. The importance of mTOR activity for articular cartilage destruction in OA is confirmed by significant changes in the work of mTOR regulatory network that involves multiple intracellular (growth factors, adenosine triphosphate, oxygen availability, and autophagy) and extracellular (glucose, amino acids, lipids, and hexosamine) signals. Moreover, the altered expression of the mTOR gene in the blood of patients with OA is associated with either increased pain or synovitis, which indicates that there is a strong metabolic heterogeneity in patients with OA and a need for a differentiated therapeutic approach. The above problems are discussed in this review.

Background. Metabolic changes in terms of developing diabetes, especially hyperglycemia, can directly affect immune metabolism of lymphocytes. Objective: The aim was to find out the level of mRNA gene expression of Glut1, mTOR and AMPK1α in pancreatic lymphatic nodes (PLN) in rats with experimental streptozotocin-induced diabetes mellitus (ESIDM) and after introduction of metformin. Methods. Glut1, mTOR and AMPK1α mRNA expression were analyzed by real-time reverse transcriptase-polymerase chain reaction. Total RNA was extracted from PLN tissue by Trizol RNA Prep 100 (Isogen, Russia), according to the manufacturer's instructions. RNA was re-suspended in RNase free water, quantified and subjected to RT-PCR reaction using RT-PCR kit; RT (Syntol, Russia). To determine the level of Glut1, mTOR and AMPK1α mRNA, RT-PCR was performed in real-time by thermal cycler CFX96 ™ Real-Time PCR Detection Systems (Bio-Rad Laboratories, Inc., USA). The relative level of gene expression were studied with rat reference genes GAPDH by the method ΔΔCt. Statistical analysis were conducted using available software «Bio-Rad SFX Manager 3.1» (Bio-Rad, USA). Results. It was established that hyperglycemia caused the transcript induction of genes of glucose transporter Glut1 (in 9,9-28,9 times, p

Objective: to study the pattern of impaired regulatory mechanisms of the mammalian target of rapamycin (TOR) signaling pathway, by monitoring gene expression in the blood of patients with osteoarthrosis (OA) at different stages of the disease. Subjects and methods. The study covered 33 outpatients with OA, 14 patients with this condition prior to knee joint endoprosthesis, and 27 healthy individuals (controls) (mean age 58.0+7.4, 56.5+8.9, and 55.0+8.3 years, respectively). Total RNA was isolated from their blood and used to determine the level of gene expression by a real-time polymerase chain reaction for AMP-activated protein kinase (AMPK), hypoxia-inducible factor-1α (HIF1α), the rate-limiting proteins of the hexosamine signaling pathway — glutamine-fructose-6-phosphate amidotransferase and acetylglucosaminyltransferase, as well as the glucose transporter GLUT1 and steps 6 and 7 glycolytic pathway components — glucose-6-phosphate dehydrogenase and phosphoglycerate kinase-1, respectively; the lipogenesis-related genes — fatty acid synthase (FAS) and the activity of the pentose phosphate pathway — glucose-6-phosphate dehydrogenase in the blood of patients with OA at different stages of the disease. Results. Analysis of gene expressions showed that in the OA patients with a low expression of the mTOR gene (a LOW subgroup), the expression of AGT and GLUT1 genes proved to be significantly lower and that of the AMPK gene was higher than in the healthy individuals. In the OA patients with a high expression of the mTOR gene (a HIGH subgroup), the expression of all the genes under study was much higher, except for the FAS gene; moreover, the greatest expression excess as compared to the controls was observed for the AMPK and HIFlα genes. In the patients with endstage disease (an ES subgroup), the expression of all the study genes, including the FAS gene, turned out to be higher than in the healthy individuals. Conclusion. The development of OA is accompanied by a considerable decrease in the efficiency of energy metabolism. At the same time, in the patients with a low mTOR gene expression, energy deficiency may be due to decreased cellular metabolite transport. It may be caused by the deficiency of the end electron acceptor oxygen in the patients with a high mTOR gene expression and the pathological redistribution of energy substrate in favor of lipogenesis cannot be ruled out in those with end-stage disease.

Objective: to study the expression ratio of the non-tissue specific regulatory genes mTOR, р21, ATG1, caspase 3, tumor necrosis factor-а (TNF-а), and interleukin-6 (IL-6), as well as matrix metalloproteinase 13 (MMP-13) and X type collagen (COL10A1), cartilage resorption-associated MMP13 and COL10A1 in the blood and knee articular cartilage in patients with rheumatoid arthritis (RA). Subjects and methods. Twenty-five specimens of the distal femoral articular cartilage condyles were studied in 15 RA patients (mean age 52.4+9.1 years) after endoprosthetic knee joint replacement and in 10 healthy individuals (mean age 36.0+9.1 years) included into the control group. Twenty-eight blood samples taken from 28 RA patients (aged 52+7.6 years) prior to endoprosthetic knee joint replacement and 27 blood samples from healthy individuals (mean age 53.6+8.3 years; a control group) were also analyzed. Real-time quantitative polymerase chain reaction was applied to estimate the expression of the mTOR, p21, ATG1, caspase 3, TNF-а, IL- 6, COL0A1, and MMP-13 genes. The levels of a protein equivalent in the p70-S6K(activated by mTOR), p21, and caspase 3 genes concerned was measured in the isolated lymphocyte lysates, by applying the commercially available ELISA kits. Total protein in the cell extracts was determined using the Bradford assay procedure. Results. The cartilage samples from patients with end-stage RA exhibited a significantly higher mTOR, ATG1, p21, TNFа, MMP-13, and COL10A1 gene expressions than did those from the healthy individuals. At the same time, IL6 gene expression was much lower than that in the control group. The expressions of the mTOR, ATG1, p21, TNFа, and IL 6 genes in the blood of RA patients were much greater than those in the donors. Caspase 3 expression did not differ essentially in the bloods of the patients with RA and healthy individuals. The bloods failed to show MMP-13 and COL10A1 expressions. High mTOR and p21 gene expressions were accompanied by the elevated concentrations of the corresponding proteins in the cell lysates of the patients with RA compared to the controls. Conclusion. The findings suggest for the first time that regulatory mTOR, ATG1, p21, and TNFа gene expressions are enhanced in the blood and articular cartilage of RA patients. These changes are accompanied by the increased expression of MMP 13 gene that is responsible for articular cartilage resorption. Therefore, the higher expression of the examined regulatory genes in the blood of RA patients may be indicative of articular cartilage degradation.

Objective: To study whether the course of the disease can be predicted in patients with osteoarthrosis (OA), by monitoring mTOR (Mammalian Target Of Rapamycin) gene expression in their blood. Material and methods. The investigation was conducted on the peripheral blood samples from 33 outpatients (58.4±7.4 years) with OA; 10 patients (56.5±8.9 years) before endoprosthetic knee joint replacement, и 27 healthy individuals (55.6+8.3 years) who formed a control group. Total RNA was isolated from the blood and used to estimate the gene expression of mTOR, autophagy-related protein 1 (ATG1), the cyclin-dependent kinase inhibitor p21, caspase 3, and tumor necrosis factor-а (TNF-а) by real-time polymerase chain reaction. Results. Analysis of gene expression in the outpatients with OA identified two subgroups: in one subgroup (n = 13) mTOR expression was considerably much less than that in the control group; the expression of ATG1 and p21 did not differ greatly from the control and that of caspase 3 and TNF-α was significantly higher. The other outpatients (n = 20) and all the examined patients needing endoprosthetic replacement were ascertained to have a higher gene expression of mTOR, ATG1, p21, caspase 3, and TNF-α than in the control group. Before endoprosthetic replacement, severe joint destruction in patients with OA was associated with enhanced gene expression of mTOR, ATG1, p21, and caspase 3. Conclusion. In early-stage disease, increased mTOR gene expression may serve as a prognostic marker of the severity of the disease and articular cartilage destruction.

Objective: to study an association of bone mineral density (BMD) with the expression of genes associated with the proliferation, survival, and differentiation of osteocytes in the peripheral blood of postmenopausal women with osteoporosis (OP). Subjects and methods. Twenty-eight postmenopausal women with OP and 17 age-matched healthy women were examined. BMD was measured by X-ray absorptionmetry. Alkaline phosphatase activity and calcium and phosphorus levels in the blood were determined by conventional methods. RNA was isolated from the peripheral blood and used to estimate the gene expression, by using real-time polymerase chain reaction (RT-PCR). Results. The postmenopausal women with OP were observed to have a significant increase in the expression of ATG1 (serine-threonine kinase that is responsible for the generation of autophagic vacuoles) as compared to the healthy controls, which suggests that autophagy develops in the blood cells of these patients. This was attended by a considerable reduction in the expression of the mTOR (the mammalian target of rapamycin) gene, a regulator of protein synthesis and cell proliferation, and in that of the genes associated with the differentiation of osteoblasts: transforming growth factor β 1 (TGF- β 1), Runx 2 (Runt-related transcription factor 2) and total alkaline phosphatase (TAP), and the proinflammatory cytokines tumor necrosis factor- α (TNF- α ) and interleukin 1 β (IL-1 β ). The expression of mTOR, TGF- β 1, Runx2, and IL-1 β genes was positively correlated with BMD in individual areas of the hip. On the contrary, that of ATG1 and TAP was correlated negatively with BMD in this area and positively with that in the spine. Although the expression of the genes in question was not correlated with the serum levels of phosphorus and calcium, that of TGF- β 1 was positively related to TAP activity in postmenopausal women with OP. Conclusion. In postmenopausal women with OP, bone loss is accompanied by a significant increase in the expression of the autophagy marker ATG1 and by a reduction in that of mTOR, which may be indicative of the inhibition of protein biosynthesis and cell proliferation and the suppression of the expression of the genes associated with the differentiation of osteoblasts and the proinflammatory cytokines TNF- α and IL-1 β . The obtained data on gene expression may be used to search for new disease markers tested in peripheral blood.