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Project titleMolecular basis for future medicine
Project LeadMakarov Alexander
AffiliationEngelhardt Institute of Molecular Biology, Russian Academy of Sciences,
|Implementation period||2014 - 2018|
Research area - , -
“Targets for cancer biotherapy” The problem of efficient cancer therapy, especially in the cases complicated with metastases, remains unsolved. Cytostatic drugs with limited specificity and expressed toxicity still form the basis for metastatic cancer therapy. The identification of specific targets and points of vulnerabilities of cancer cells, and the development of approaches for their application in the therapy are among the most current objectives of biomedicine. The emerging key issues of the studies involve elucidation of the most common hallmarks of cancer cells, such as the loss of tumor suppressor p53 function, deficient antiviral defense mechanisms, and dysfunction of mitochondrial respiration, as well as finding of some tissue-specific targets that would allow affecting cancer cells based on their tissue origin. The loss of the p53-dependent sorting-out of abnormal cells is found in virtually all cancer cases. However recently the research team of the Engelhardt Institute of Molecular Biology followed by other research groups, has discovered homeostatic activities of p53 that help in better accomodation of the cells subjected to physiological stresses. To find the most reliable therapy choice it is important to know to what extent can the homeostatic activities interfere with cytostatic anticancer therapy. One of the approaches to solve the problem in the planned studies will use oncolytic viruses as biotherapeutic agents. Such viruses are able to kill cancer cells selectively without any effect on normal cells. The primary objective is to provide the safety and predictability of therapy based on the oncolytic viruses. The mechanisms that are responsible for susceptibility of cancer cells to the oncolytic viruses will be studied. The problem of acquired resistance of malignant cells to cancer therapy will be addressed through the use of notorious plasticity of the genome of cancer cells. As a result of genetic competition the oncolytic virus would adapt to changing cells faster than cancer cells would acquire the resistance to the virus. Exploration of new specific targets for therapy of cancer cases that originate from the cells of certain tissue, is important for the development of new instruments for fighting malignancies. One of the proteins capable to serve as such a target is MCTS1 oncoprotein that participates in translational regulation. Recently we found that the MCTS1 gene expression is altered during the genesis of human lymphomas. The therapeutic potential of MCTS1 inhibitors as anti-limphoma agents will be studied in the course of the project. One of the important tasks of modern biomedicine is related to studies on non-coding RNAs ( ncRNA) and their role in gene regulation. The determination of the role of the long ncRNA repertoire change in the case of cancer is as much important as the search of MCTS1 inhibitors. On the one hand, it will allow conducting quick cancer diagnostics by serum RNA analysis, which is secreted by tumor cells by exosome secretion, and on the other hand, it will give the chance to determine special characteristics of the tumor of the certain patient for the optimal therapy course. The colon cancer repertoire of long ncRNA will be studied and the serodiagnostic assay based on analysis of long ncRNA from patients will be developed. Each task in this section is characterized by conceptual novelty and is based on unique pilot projects and approaches suggested by members of the scientific team. Homeostatic functions of p53 were originally discovered in EIMB. The connection of losing the antiviral mechanisms to losing the p53 tumor suppressor functions was not studied previously. The idea of using the genetic competition for overcoming the acquired resistance of tumor cells to therapy is entirely innovative and was not proposed by anybody else earlier. The team of applicants proposes the study of the possibility of using the MCTS1 oncoprotein as therapeutic target for the first time as well. Finally, the use of the analysis of long ncRNA repertoire for the personalized diagnostics is also based on original projects of the team and possesses novelty. Scientific results, which will be obtained in the course of investigations, will be of the highest possible standard and correspond to the vital needs of the medicine. "Cytokines as a key to understanding the mechanisms and treatment of diseases" Cytokines are not only important regulators of immunity, they also belong to the extracellular components of chronic inflammatory reactions leading to a variety of human diseases. This makes cytokine research highly relevant. New drugs introduced in recent years and aimed at blocking certain cytokines under various pathological conditions associated with inflammation, have opened new horizons in immunology, oncology and other fields of medicine. These drugs also revolutionized both the clinical medicine and the global pharmaceutical industry. However, despite this remarkable progress, the understanding of the molecular and cellular mechanisms of cytokine action in disease remains rather incomplete, which leads to unwanted side effects. The proposed program section aimed at creating new experimental models of various diseases, including inflammatory and autoimmune disorders as well as cancer, and studying the functions of cytokines, is based on many years of research carried out in collaboration with the world's leading experts in cytokine biology. We are going to use recently created unique strains of genetically modified mice, as well as new mouse models to be generated in the course of the project. In particular, we plan to generate a new panel of mice with "humanization" of proinflammatory cytokines, where genes for both cytokines and their receptors will be replaced in the mouse genome by corresponding human genes. Such mice will be suitable to study the effects of drugs developed for the clinic, which are inactive in normal mice. We will focus on experimental models of human pathologies associated with pro-inflammatory cytokines. Furthermore, panels of mouse strains with inactivation of the individual cytokine genes to specific cell types will be created. Taken together, these experimental models will help to establish the molecular mechanisms of disease development under the influence of cytokines, that is, to identify new molecular links between cytokine signaling cascades and diseases, and to find new approaches to their therapeutic blocking. Molecular mechanisms that explain the contribution of chronic inflammation in the development of cancer will be studied in detail. “Development of drug prototypes, targeting host cell components, for the treatment of socially significant infectious diseases” Socially significant diseases, including HIV and HCV-induced diseases, affect the essential part of the population and greatly influence different aspects of social life. A number of drugs, which effectively block viral life cycles, are developed to date. These drugs generally affect components of the virus. Genetic variability of the pathogens causes rapid emergence of resistant strains. This, in turn, requires frequent drug replacement and increases the therapy cost. The problem still remains of vital importance. Scientific novelty and significance of the task comprise new approach to the development of pharmaceuticals, acting on different targets including host cell components, which directly or indirectly participate in the pathogenesis. The major advantage of this approach is the absence of drug resistance. Host cell proteins, which are involved in HIV and HCV life cycles are studied insufficiently, and drugs affecting such proteins have not been introduced yet. The implementation of such approach requires detailed study of the fundamental aspects of the host-pathogen interaction. To date, we synthesized a variety of original compounds, which inhibit HIV and HCV life cycles in cellular systems. Mechanisms of their action and host-cell targets are still unrevealed. Determination of such targets and the optimization of molecules will help to develop prototypes of new antiviral drugs. The development of new generation of safe and effective vaccines is important for healthcare and veterinary medicine. The drugs based on DNA-vaccine technology are being actively designed at the moment. Thus, four DNA-vaccine medications are already presented in the world market, and dozens of medications are in the clinical trial stage. The potential of the given type of vaccines may be increased significantly by means of genetic engineering modifications of the DNA-vaccine antigens. The novelty of our approach to the construction of DNA-vaccines lies in the antigen modification by signal peptides, which makes it possible to form the effective protective immunity in the organism. Current situation in the tuberculosis therapy calls for radical replacement of the drugs in use since most of them are not active against ubiquitously spread strains with multiple drug resistance and extensive drug resistance. The resource of the four first-line medications – isoniazid, rifampicin, pyrazinamide, and ethambutol, – which were introduced into practice more than 40 years ago, is considerably depleted. This situation requires search for the new targets among the M. tuberculosis proteins and development of compounds affecting them specifically. The scientific novelty and significance of the proposed research lies in the development of new anti-tuberculosis (anti-TB) drugs based on the modified nucleosides. To date compounds of this class have not been used as anti-TB remedies. In our preliminary experiments, a number of such compounds possess significant antimycobacterial activity, including anti-MDR activity. This approach is unique due to the availability of a number of compounds synthesized at the EIMB RAS, in part patented in Russian Federation. The high-technology products, which will be created as a result of this research, will have high potential for commercial use in Russia and abroad. “Molecular methods of multiplex analysis of the protein and DNA markers of socially significant diseases” (i) Development of a multiplex method for determining the profiles of different immunoglobulin classes for molecular diagnosis of allergic diseases. Allergic diseases are accompanied by changes in the profiles of different classes of immunoglobulins (IgE, IgG, IgM, and other) responsible for the onset and specific progression of allergic disorders. To understand the pathogenesis of an allergic condition it is necessary to accurately determine the levels of allergen-specific immunoglobulins that undergo changes, which requires testing tens and sometimes hundreds of allergens for their effect on the organism. Subsequently, during immunotherapy it is required to monitor the profiles of different classes of immunoglobulins to evaluate changes in their levels, showing the degree of adequacy of the treatment. At present the universally accepted technique applied to detect the allergens causing allergic condition is the intradermal injection of allergens, followed by visual inspection of the size of the inflammation area, which is traumatic and unsafe for the patient. The in vitro methods used as an alternative do not allow simultaneous testing for the total IgE, and individual sIgE and sIgG4 against a set of natural allergens of different groups in a single sample of blood serum. Scientific novelty and relevance of the task lies in creating a new approach to the diagnostics and monitoring of the treatment of allergic diseases, based on multiplex testing of the patient's sensitivity to a wide range of allergens, and determining the profiles of over a hundred different classes of immunoglobulins. The resulting information on the ratio of different immunoglobulin classes will allow identifying the individual mechanism of an allergic condition for each patient. Uniqueness of this approach is substantiated by the use of hydrogel protein biochips as an analytical tool. The biochips are produced utilizing the technology developed and patented at the EIMB RAS. The biochip technology provides a hydrophilic environment for the proteins immobilized in the biochip cells and preserves the protein initial activity, resulting in high sensitivity, reproducibility, and stability of the biochip-based test systems if stored for an extended period. (ii) Personalized microsystem for the analysis of pathogenesis of Mycobacterium tuberculosis. Recent research in genomics and epidemiology of Mycobacterium tuberculosis showed that rapid increase in the number of strains with various resistance profiles (poly-resistant, multi-resistant, and extensively and super extensively drug-resistant forms) is caused by the uncontrolled one-time application of a spectrum of the first- and second-line anti-TB drugs, which results in the Mycobacterium developing effective defense mechanisms. New generation of drugs with proven antibacterial effect (perhlozon, bedakvilin, lineazolid, nitroimidazoles), which are being already used for practical application or are in the final stages of clinical trials, are designed to overcome the existing problems of treating patients with drug-resistant forms of TB. However, their effectiveness will depend on the use of molecular diagnostic tools based on the knowledge of specific genetic determinants of resistance and other markers with prognostic value for the disease progression and for personalized treatment. If for the first- and second-line anti-TB drugs the mechanisms of resistance have been well studied, for the new generation third-line drugs such data are currently unavailable. This line of research aimed at establishing the determinants of resistance to the new powerful anti-TB drugs, as well as at analyzing the clinical significance of genetic factors in the pathogenesis of various Mycobacterium tuberculosis families, will be based on full genome sequencing of the collections of clinical isolates of M.tuberculosis. Using the obtained data a new screening diagnostic tool will be developed based on the hydrogel biochip technology for comprehensive analysis of the Mycobacterium tuberculosis genome, allowing to perform effective personalized treatment and prevent the emergence and spread of new highly virulent drug-resistant forms of TB. The high-technology products developed as a result of this project will be highly commercially viable in Russia and for export. “Basic mechanisms underlying human health and longevity” Many eukaryotic organisms (e.g. sponges, flat worms and hydra) do not exhibit any manifestations of aging process. Furthermore, the lack of senility is a characteristic feature of several mammalian species including naked mole rat and certain bat species. There exist a few mammalian species with life span twice as long as the human life span. Besides, at the present time there are multiple examples of experiments that extended the duration of life span several fold using genetic approaches, low calorie diet and other interventions. In particular the normal life span of C. elegans was extended 10-fold. Such experiments demonstrate that it is possible at the present time to manipulate life span duration exploring gene therapy or pharmacological approaches. (i) Therefore, it is of great importance to investigate peculiarities of the structure and function of genomes of the D. melanogaster strains that differ drastically with respect to the duration of life span, as well as of grey whale (Eschrichtius robustus), one of the most long-lived species among the mammals. Comparative analysis of the transcriptomes of these phylogenetically distant organisms will shed light on the highly conserved gene-candidates, which are responsible for longevity, and will allow to develop the Drosophila transgenic strains, containing these genes. (ii) The importance of planned investigation of molecular mechanisms underlying DNA repair system stems from the vital role of this system function in the process of aging and senility formation. It was demonstrated that phenomenon of premature senility results from genetic impairments of various DNA repair systems including the transcription-coupled repair (TCR) system which plays the key role in this process. The proposed detailed analysis of the recently discovered new molecular mechanism of TCR, providing coordination between transcription and DNA repair systems will be carried out using bacteria as a model, coupled with cloning and description of repair genes in Drosophila. Results of this analysis should help to reveal new pathways controlling the function of DNA repair-related enzymes, that may protect organisms from premature aging and the resulting senility. (iii) The duration of human life span and healthy life is very limited due to bad ecology in the cities and constant extreme psychological stress. Therefore, a search for non-toxic organic agents that may help to decrease the level of oxidative stress and, hence, extend the duration of healthy and creative period of human life is of great urgency at the present time. It is well known that after various types of stress the induction of several stress-proteins is observed with Hsp70 apparently playing the major protective role. This protein is a universal adaptogene which ameliorates the deleterious consequences of several proteinopathies including the level of oxidative stress. Therefore, the major goal of our research is to develop a new anti-stress drug basing on recombinant human Hsp70 which will be produced using various prokaryotic and eukaryotic expression systems. The administration of this drug should increase working capacity and extend creative period of human life.
"Targets for cancer biotherapy" Priority results will be obtained to advance novel approaches to personalized cancer biotherapy. Individual therapy approaches will be developed for cancer cases associated with the preserved homeostatic functions of wild-type p53. The homeostatic functions of p53 are mediated by a set of p53-regulated genes that will be used as new therapeutic targets. The development of a personalized use of oncolytic viruses will allow choosing optimal for the particular patient therapeutic strains. The development of new oncolytic strains affecting a wide range of tumors will overcome the problem of an acquired resistance to the therapy. A screening for inhibitors of the MCTS1 oncoprotein will be carried out both in vitro and in vivo using a library of small molecules that potentially block the interaction of MCTS1 with DENR or with ribosomal of tRNAs. Specific mRNA targets of the MCTS1 oncoprotein will be identified that will allow developing approaches to treatment of the MCTS1 gene amplification provoked lymphomas with already certified drugs. Identification of specific repertoires of long ncRNA secreted into the serum through exosomes from the colon tumor will make it possible not only to develop the rapid diagnostic PCR test but will also allow to reveal individual features of the patient tumor providing reference points for the personalized therapy. "Cytokines as a key to understanding the mechanisms and treatment of diseases" The series of studies establishing the contribution of TNF from various cell types to the development of experimental arthritis, as well as to the tuberculosis pathogenesis, will be brought to completion. We will measure the expression profile of cytokines in models of acute and chronic skin inflammation and determine the contribution of these cytokines to skin carcinogenesis. The role of the signaling axis composed of membrane lymphotoxin and its receptor in chemically-induced colitis and experimental intestinal ischemia will be determined. An experimental platform to study the effects of anti-cytokine therapy, including the effects on microbiota, will be developed using "humanized" mice. New mouse models with cell-type-specific inactivation of IL-1a, IL-1b and RelB component NFkB transcription factor will be used to determine the role of these molecules in oncogenic inflammation of the intestine. In collaboration with pharmaceutical companies, new mouse models of human disease created in the course of the project will be used to develop innovative anti-cytokine drugs and to radically improve the prognosis during anti-cytokine therapy. “Development of drug prototypes, targeting host cell components, for the treatment of socially significant infectious diseases” The functional role of some host-cell proteins in the HIV and HCV life cycles will be established. The compounds – prototypes of antiviral drugs, which bind specifically to these proteins and inhibit viral replication, will be synthesized. The use of such agents will not cause the emergence of drug resistance. It will allow significant simplification of the therapy and reduction of its cost. The prototypes of DNA-vaccines against human and animal viral infections (tick-borne encephalitis, parainfluenza and virus diarrhea) will be developed. There is a high demand for such agents in healthcare and veterinary medicine. New drugs based on modified nucleosides will be developed, and their targets in M. tuberculosis will be evaluated. These compounds will represent the prototypes of anti-tuberculosis drugs, which will equally affect wild-type M. tuberculosis and the strains with multiple drug resistance. At the last stage of this work, all prepared drugs will be submitted for preclinical and clinical trials to the corresponding specialized institutions, including Central Tuberculosis Research Institute RAMS, Chumakov Institute of Poliomyelitis and Viral Encephalitides RAMS, Ivanovski Research Institute of Virology RAMS, Federal State Unitary Enterprise «Kurskaya biofabrika – BIOK» and also to commercial pharmaceutical companies. The pharmacodynamic and pharmacokinetic properties of drugs, which will be developed, should meet internationally accepted standards and correspond to scientific trends in this field. Patents for drugs will be obtained and could be used to substitute the imported medications. “Molecular methods of multiplex analysis of the protein and DNA markers of socially significant diseases” A method of the multiplex immunofluorescence assay on hydrogel biochips will be developed to analyze the profiles of allergen-specific immunoglobulin classes sIgE and sIgG4. A diagnostic test system will be developed on the basis of this method for parallel quantification of not less than 60 sIgE and 60 sIgG4 for different classes of allergens (pollen, house dust, epidermal, food, and fungal allergens, insect venoms), as well as determining the concentration of total IgE, performed in a single test of serum on the hydrogel biochip. The recombinant allergens as well as protein extracts isolated from natural sources will be immobilized within the hydrogel biochip cells. Parallel testing of immuno-complexes of the immobilized allergens bound to different classes of antibodies will be performed using a mixture of the antihuman IgE- and IgG4- specific antibodies conjugated to different fluorescent dyes. A customized test system on the hydrogel biochip will be created allowing to identify DNA of the tuberculosis causative agent, determine its genotype and identify the clinically significant factors in its pathogenesis, including mutations that determine resistance to perhlozon, bedakvilin, lineazolid, nitroimidazoles, as well as mutations that compensate for the reduction of bacterial fitness and fix the stable state of the strain. The biochip serving as a key detecting element of the microsystem, will contain an integrated personalized electronic identifier, providing access to the data about the patient and results of the clinical sample tests. A hardware system will be developed to perform automated processing and storage of the tests results, and data exchange with the in-house database connected to the local hospital network. At the final stage of the project the two systems will be submitted for approval to the Federal Service on Surveillance in Healthcare as medical devices for in vitro diagnostics. For the purposes of conducting clinical trials, the following medical organizations will be involved: Clinic No 1 of the Office of the President of the Russian Federation, State Scientific Centre “Institute of Immunology, Federal Medical and Biological Agency”, and the anti-TB centers of the Russian Ministry of Health. Technical documentation describing the systems will be transferred to a subsidiary of EIMB RAS, LLC "Biochip-IMB" for production and commercialization of the test systems. The proposed systems are user-friendly and they allow informative screening for laboratory diagnostics of allergy and tuberculosis. The systems should meet internationally accepted standards and scientific trends in this field. Application of these systems in routine practice of laboratory services will yield substantial socio-economic benefits through enabling provision of an effective personalized therapy, shorter periods of disability, hospitalization and treatment at the early stages of the disease. “Basic mechanisms underlying human health and longevity” Deep sequencing of the transcriptomes of D. melanogaster strains that differ drastically in terms of longevity will reveal the putative gene-candidates that may significantly affect this parameter. Sequencing of the long-lived grey whale and assembly of the resulting contigs will enable us to compare the results with the available relevant data accumulated previously by the authors of the project when studying other long-lived organisms. The obtained results will allow to develop the Drosophila transgenic strains containing these candidate-genes controlling longevity in their genomes. Another goal of the experiments will be to block transpositions of retroelements in the brain of model organisms (rats) with the help of shRNAs and monitor the effect of such blockade on longevity. Utilizing the experimental system developed by us, we will investigate the role of helicase UvrD in different types of DNA repair systems and perform a search for new regulatory proteins and metabolites that may prolong normal activity of DNA repair systems in the process of aging. Additionally, new approaches will be developed to protect organism from the deleterious consequences of oxidative stress, on the basis of directed modification of the DNA excision repair system coupled with transcription (TCR). This investigation will provide a list of putative signal molecules with positive effects on the life span due to enhanced activity of DNA repair systems. Finally, the realization of this project will shed light on various protective and chaperone characteristics of the human recombinant Hsp70 samples. Within the framework of the project, preclinical testing of the human Hsp70 will be carried out to evaluate its potential as a pharmaceutical anti-stress drug.
Annotation of the results obtained in 2018
"Cytokines - the key to understanding the mechanisms and therapy of deseases" The most significant result in 2018 was the discovery of a new protective function of TNFR2 on T-regulatory cells in the pathogenesis of experimental autoimmune encephalomyelitis (EAE). Here, for the first time we show the significance of the TNF/TNFR2 signaling cascade for the maintenance of immunosuppressive function of T-regulatory cells in vivo. Moreover, these results may explain the inefficiency of anti-TNF therapy in the treatment of multiple sclerosis. To directly address the role of TNFR2, during the past several years we developed and validated a doubly humanized TNF/TNFR2 mouse line. In addition, we performed a comparative analysis of several VHH-based bispecific antibodies against hTNF with various specificities to the surface markers on myeloid cells to obtain the most effective blocker. In collaboration with two international groups, the nonredundant protective function of TNF from basophils was discovered in the experimental model of polymicrobial sepsis, and TNF from mast cells was shown as an activator of the immune response in the LPS-induced dermatitis model. Finally, in an experimental model of severe allergic asthma the same cytokine produced by macrophages and dendritic cells differently contributed to the development of eosinophilic and neutrophilic inflammation in the lungs, respectively. The main results of our project are essential for the understanding of how cytokines are involved in the pathogenesis of various diseases and maintenance of immunity, but also provide new approaches to the treatment of multiple sclerosis, asthma, and other autoinflammatory and infectious diseases. "Targets for cancer biotherapy" It has been established that the unmutated copies of the p53 gene that are present in some malignant cells have a protective effect against a cytotoxic chemotherapy, by activating the process of autophagy and inhibition of the mTORC1 complex. A recombinant strain of oncolytic vaccinia virus has been obtained that contains a polycistronic cassette for the expression of the cytokine GM-CSF and single-chain VHH antibody to the CD47 protein. As a result of the panoramic proteomic analysis, individual changes in the expression of the protein components of the interferon response system were identified in eight human glioblastoma cell lines. Using a bioselection in the presence of RNA mutagens, oncolytic enteroviruses with extended tropism against tumor cells were obtained. A fundamental breakthrough has been achieved in understanding the function of the MCTS1 oncoprotein and the molecular mechanism of the MCTS1 / DENR heterodimer. This achievement is extremely important in terms of the prospects for treating a wide class of lymphomas associated with an increased level of expression of the MCTS1 oncogene or the activity of its product, MCTS1 oncoprotein. We have discovered enhancer lncRNA LINC00973, which is an informative prognostic marker of progression and relapse of various types of tumors, as well as a promising target for their chemotherapy. Besides, we have identified mRNA AKR1B10 – a new powerful marker for serological diagnosis of colon cancer, whose content is strongly decreased in sera of colon cancer patients, as compared to healthy donors. "Basic mechanisms underlying human health and longevity" Full-genome sequencing of transcriptomes from long-lived Drosophila melanogaster flies with overexpression of the Gclc gene revealed changes in signaling pathways associated with the metabolism of starch and sucrose, the metabolism of several drugs, xenobiotics, glutathione and neuroactive ligand-receptor interaction. Analysis of the results of deep sequencing of the transcriptome of Drosophila melanogaster specimens heterozygous for the mutation in the E (z) gene demonstrated suppression of the genes involved in the metabolism of ascorbate and aldarate, retinol, porphyrins, xenobiotics, serine and glycine. The mechanism of interaction of UvrD protein with RNA polymerase was elucidated. Mutations in the regions encoding the C-terminal domain of UvrD, as well as a short insertion of the RNA polymerase in the beta-1 domain, have been shown to increase the sensitivity of cells to genotoxic agents. The data obtained indicate the conservatism of the proposed mechanism of DNA repair in bacteria. It has been shown that inactivation of ppGpp binding sites with “1” and “2” RNA polymerase sites involved in the regulation of transcription initiation, does not affect the influence of ppGpp on UvrD-dependent bias of RNA polymerase in vivo and in vitro. Subchronic toxicity and local irritating effects of human recombinant HSP70 after intranasal administration were studied. The experiments did not reveal a pronounced toxic effect on the general condition and the main systems of the body of model animals (rats), including biochemical parameters of blood after chronic treatment. In cooperation with the group of Dr. N.V. Bobkova (IBC RAS) we performed experiments to study the effect of recombinant HSP70 on the culture of the embryonic hippocampus of different age (5 or 25 days). The protective function of HSP70 is shown. When HSP70 was added into the hippocampal tissue culture, there was a decrease in the expression of a significant number of genes involved in the inflammatory, immune response, as well as angiogenesis, cell migration, and chemotaxis. "Development of drug prototypes, targeting host cell components, for the treatment of socially significant infectious diseases" A new test system has been designed for analysis of the hepatitis C virus replication inhibitors as potential drugs. The developed system allowed to discern the antiviral and antiproliferative activities of these inhibitors. Potential cell targets for anti-HCV therapy were identified and leader compounds were synthesized. Genetically engineering DNA vaccines prototypes against tick-borne encephalitis, rabies and Lassa viruses were produced. It has been shown that viral vaccine antigens differ in their ability to cause oxidative stress and apoptosis in cells expressing them. The developed DNA vaccine prototypes induce activation of the immunity and the parameters of the latter can be further improved. Antibiotic activity of modified pyrimidine nucleotides was demonstrated against a number of gram-positive and gram-negative bacteria. The compound possessed pronounced activity against mycobacteria. It was shown that the treatment of M. tuberculosis H37Rv cells with some of the synthesized compounds lead to degradation and then destruction of the cell wall. "Molecular methods of multiplex analysis of the protein and DNA markers of socially significant diseases" ALLERGO-BIOCHIP diagnostic kit based on the microarray containing 44 immobilized allergens of various classes has been developed and validated. Technical documentation has been prepared and submitted to institution accredited by Roszdravnadzor for technical examination. Profiles of sIgE, sIgG levels, including sIgG4, and sIgA, as well as correlations of sIgG & sIgA, sIgE & sIgG4 levels in allergic diseases were investigated using biochips. The significance of simultaneous analysis of sIgE and sIgG4, sIgG, sIgA levels in allergic diseases was demonstrated. An importance of controlling the levels of these antibodies during allergen-specific immunotherapy is shown. Microbiological and genetic criteria for detecting resistance of the tuberculosis causative agent to bedaquiline and linezolid were formulated and described. It was established that the genetic determinants of resistance to these drugs are localized in the mmpR, atpE, atpB, rplC and rrl genes. In order to identify these targets, an oligonucleotide microarray and a corresponding TB-Biochip IV diagnostic kit were developed, which successfully passed preliminary trials. Based on these results technical documentation for kit production was prepared, which will be handed over to the BIOCHIP-IMB, LLC as EIMB subsidiary on terms of the license agreement after grant of сorresponding patent.
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13. Rezvykh A.P., Zatsepina O.G., Yurinskaya M.M., Vinokurov M.G., Krasnov G.S., Mitkevich V.A., Makarov A.A., Evgen’ev M.B. The Effect of beta-Amyloid Peptides and Main Stress Protein HSP70 on Human SH-SY5Y Neuroblastoma Proteome Molecular Biology, 52, 6, 1082-1092 (year - 2018).
14. Valuev-Elliston V.T., Kochetkov S.N. Novel HIV-1 Non-nucleoside Reverse Transcriptase Inhibitors: A Combinatorial Approach Biochemistry-Moscow, 82, 13,1716-1743 (year - 2017).
15. Kozlov M.V., Malikova A.Z., Kamarova K.A., Konduktorov K.A., Kochetkov S.N. Synthesis of Pyridyl-4-Oxy-Substituted N-Hydroxy Amides of Cinnamic Acid as New Inhibitors of Histone Deacetylase Activity and Hepatitis C Virus Replication Russian Journal of Bioorganic Chemistry, 44, 4, 453–460 (year - 2018).
16. Voloshin S., Smoldovskaya O., Feyzkhanova G., Arefieva A., Pavlushkina L., Filatova T., Butvilovskaya V., Filippova M., Lysov Y., Shcherbo S., Makarov A., Rubina A., Zasedatelev A. Patterns of sensitization to inhalant and food allergens among pediatric patients from the Moscow region (Russian Federation) PLoS ONE, 13, 3, e0194775 (year - 2018).
17. Gubernatorova E.O., Gorshkova E.A., Namakanova O.A., Zvartsev R.V., Hidalgo J., Drutskaya M.S., Tumanov A.V., Nedospasov S.A. Non-redundant functions of IL-6 produced by macrophages and dendritic cells in allergic airway inflammation Frontiers in immunology, 9, 2718 (year - 2018).
18. Khandazhinskaya A.L., Alexandrova L.A., Matyugina E.S., Solyev P.N., Efremenkova O.V., Buckheit K.W., Wilkinson M., Buckheit Jr. R.W., Chernousova L.N., Smirnova T.G., Andreevskaya S.N., Leonova O.G., Popenko V.I., Kochetkov S.N., Seley-Radtke K.L. Novel 5'-Norcarbocyclic Pyrimidine Derivatives as Antibacterial Agents Molecules (Basel, Switzerland), 23, 12, 3069 (year - 2018).
19. Gryadunov D.A., Shaskolskii B.L., Nasedkina T.V., Rubina A.Y., Zasedatelev A.S. Технология гидрогелевых биочипов ИМБ РАН: 30 лет спустя Acta Naturae, 10, 4, 30-44 (year - 2018).
20. Smoldovskaya O., Feyzkhanova G., Voloshin S., Arefieva A., Chubarova A., Pavlushkina L., Filatova T., Antonova E., Timofeeva E., Butvilovskaya V., Lysov Y., Zasedatelev A., Rubina A. Allergen-specific IgE and IgG4 patterns among patients with different allergic diseases World Allergy Organization Journal, 11, 35 (year - 2018).
21. Zimenkov D.V., Nosova E.Y., Antonova O.V., Krylova L.Y., Peretokina I.V., Kholina M.S., Safonova S.G., Gryadunov D.A. Genomic Diversity of clinical Mycobacterium tuberculosis Isolates with Altered Susceptibility to Bedaquiline Clinical Infectious Diseases, - (year - 2018).
22. Nosenko M.A., Atretkhany K.N., Mokhonov V.V., Vasilenko E.A., Kruglov A.A., Tillib S.V., Drutskaya M.S., Nedospasov S.A. Modulation of bioavailability of proinflammatory cytokines produced by myeloid cells Seminars in Arthritis and Rheumatism, - (year - 2018).
23. Piliponsky A.M., Shubin N.J., Lahiri A., Truong P., Clauson M., Niino K., Tsuha A., Nedospasov S.A., Karasuyama H., Reber L.L., Tsai M., Mukai K., Galli S.J. Basophil-derived TNF can enhance survival in a sepsis model in mice Nature Immunology, - (year - 2018).
24. Yanvarev D.V., Kalnina L.B., Korovina A.N., Solyev P.N., Karpenko I.L., Kukhanova M.K., Nosik D.N., Kochetkov S.N. Влияние ингибиторов мевалонатного пути биосинтеза холестерина на репликацию ВИЧ в лимфоцитах человека Acta Naturae, - (year - 2019).
25. Evgen’ev M.B., Garbuz D.G., Morozov A.V., Bobkova N.V. Intranasal Administration of Hsp70: Molecular and Therapeutic Consequences Heat Shock Proteins: HSP70 in Human Diseases and Disorders, Springer International Publishing AG, part of Springer Nature 2018, Cham, Switzerland, 14, 305-323 (year - 2018).
26. - Грантополучатели РНФ расскажут о своей работе и проведут экскурсии по лабораториям во время Всероссийского фестиваля науки NAUKA 0+ Сайт РНФ, 02.10.2018 (year - ).
27. - Раскрыт механизм работы связанных с раком и аутизмом белков Индикатор, 24.08.2018 (year - ).
28. - Российские противоопухолевые препараты на основе онколитических вирусов могут появиться в течение двух-трех лет (2) Сайт РНФ, 21.09.2018 (year - ).
29. - Ученые изучили молекулу, блокировка которой поможет в лечении аллергической астмы Сайт РАН, 13.12.2018 (year - ).
30. - Грантополучатели РНФ рассказали о своих исследованиях на конгрессе по аутоиммунным и иммунодефицитным заболеваниям Сайт РНФ, 29.11.2018 (year - ).
31. - Российские противоопухолевые препараты на основе онколитических вирусов могут появиться в течение двух-трех лет (1) Medvestnik.ru, 20.09.2018 (year - ).
32. - Найдена новая мишень для лечения аллергической астмы Индикатор, 13.12.2018 (year - ).
33. - Белок TNF помогает противостоять рассеянному склерозу Полит.ру, 05.12.2018 (year - ).
34. - Российские ученые раскрыли секрет «неуязвимости» рассеянного склероза РИА Новости, 05.12.2018 (year - ).
35. - Ученые выяснили, как фактор некроза опухоли может защищать от воспалительных заболеваний нервной системы (1) Сайт РНФ, 05.12.2018 (year - ).
36. - Ученые выяснили, как фактор некроза опухоли может защищать от воспалительных заболеваний нервной системы (2) Газета.ru, 05.12.2018 (year - ).
37. - Институт молекулярной биологии РАН открыл механизм появления аллергической астмы ГлобалМСК.ру, 13.12.2018 (year - ).
38. - Российские биологи сделали открытие, которое поможет избавиться от астмы Информационный портал New Inform, 13.12.2018 (year - ).
39. - В Институте молекулярной биологии РАН рассказали о результатах масштабного исследования природы социально-значимых заболеваний Сайт РНФ, 28.09.2018 (year - ).
40. - Отредактированные мыши указали на причину неудач в лечении рассеянного склероза Индикатор, 05.12.2018 (year - ).
41. - Биологи из России раскрыли секрет появления аллергической астмы РИА Новости, 13.12.2018 (year - ).
42. - Сюжет в «Вестях» -микрочипы для аллергенов Телеканал Россия 1, 07.05.2018 Вести в 20:00 41-я минута (year - ).
Annotation of the results obtained in 2014
Annotation of the results obtained in 2015
“Targets for cancer biotherapy”. The following progress has been made within this direction of studies. We have established the involvement of p53 family members encoded by the genes TP63 and TP73 in homeostatic activities of p53. One of the genes regulated by homeostatic activities of p53, the SESN2 gene, was found to have additional regulation from the ATF4 transcription factor, which results in the viability of tumor cells under conditions of mitochondrial stress. We found that cancer cells frequently lose innate mechanisms of antiviral resistance, although cancer cells that retain wild-type p53 demonstrate better preservation of the interferon system. A treatment of HPV-positive cervical carcinoma cells with the ribonuclease binase results in an increase in the sensitivity of cells to interferon, because of an inhibition of the papillomavirus gene products E6 and E7. We have validated attainability of an adaptation of oncolytic enteroviruses to propagation within cells that were initially poorly sensitive to the virus through multiple rounds of infection in mixed cultures. The finding opens an approach to designing virus-based anticancer therapeutic substances that are not prone to the development of therapy-induced resistance. Expression constructs, assay systems and cell models have been developed for further studies of oncogenic activities of the MCTS1 translation factor. Direct interaction between products of two tumor suppressors CTDSPL and RB1 has been detected. Micro-RNAs participating in the modulation of CTDSPL gene expression have been identified. A register of long non-coding RNAs transcribed in colon carcinomas has been created. Species of long ncRNAs have been identified that that have higher levels in the cancer cells compared to the norm, as well as the species demonstrating changes in expression mirroring those of the c-MYC oncogene . A regulatory loop between the MYC gene and the HOTAIR long ncRNA has been revealed. Studies within the framework of direction “Cytokines as a key to understanding the mechanisms and treatment of diseases” are dedicated to the role of cytokines and signaling cascades released by them in the context of inflammation, autoimmune diseases and cancer, using mouse models. The effect of tumor necrosis factor (TNF) has been studied on the progression of experimental infections in mice caused by Mycobaterium tuberculosis and Mycobaterium avium. The role of TNF produced by B-cells, in the formation of B-cell infiltratrates in the lungs during tuberculosis infection has been shown. In the models of autoimmune arthritis induced by antibodies to collagen, it has been shown that mice with the inactivated TNF in macrophages are resistant to onset of the disease. Also the role has been shown of the TNF produced by mast and myeloid cells, in the induction of inflammation. The expression profiles have been determined for TNF and the related to it lymphotoxin, as well as other cytokines, in the models of acute and chronic inflammation of the skin, and the contribution of these cytokines to carcinogenesis has been established. The role of cytokine signaling pathways in infectious inflammation of the intestine caused by Citrobacter rodentium has been studied. It has been shown that genetic inactivation of the lymphotoxin in innate lymphoid cells increases the sensitivity of mice to this disease, suggesting an important role of this signaling pathway in protection against infectious colitis. New mouse models have been established featuring tissue-specific inactivation of the interleukin IL-1a and IL-1b genes, as well as transcription factor RelB playing a key role in the transmission of signals from the lymphotoxin receptor. Utilizing the "humanized" mice, an experimental platform has been developed for the study of the outcome of anti-cytokine therapy for concomitant infections. Within the framework of direction “Development of drug prototypes targeting host cell components for the treatment of socially significant infectious diseases” a range of cellular targets have been identified that mediate the effect of hepatitis C virus replication inhibitors. Among these targets are the histone acetylases, matrix metalloproteinases and retinoic acid receptor (RARb2). It has been shown that the capsid and NS5A proteins of hepatitis C virus to enhance the expression of peroxiredoxins 1, 3 and 5 in the hepatocarcinoma Huh7 cells. It has been established for the first time that methylene bisphosphonates, while showing substantial absence of cytostaticity exhibit an antiviral effect in HIV-infected MT4 cells by inhibiting the enzymes of cholesterol biosynthesis. Using the consensus sequence of NS1 protein of the Siberian subtype of encephalitis (TBE) virus, several versions of the prototype DNA vaccine against TBE have been obtained. Studies in cell culture showed that the expression of NS1 activates immune cascades in the cell. Design, synthesis and testing has been performed of modified derivatives of the nucleosides capable of specific inhibition of the growth of Mycobacterium tuberculosis, including multidrug resistant (MDR) strains. One of the physiological targets of these compounds is the tuberculosis thymidylate synthase X. Within the framework of direction "Molecular methods of multiplex analysis of the protein and DNA markers of socially significant diseases" the following results have been obtained in the development of hydrogel biochips -based molecular diagnostic systems for (i) determining personal allergic status of the patient and (ii) analysis of the pathogenesis of the causative agent of tuberculosis (TB). (i) A set of immunoreagents including allergen extracts and recombinant proteins has been collected and characterized. A comparative analysis has been conducted of the efficiency of allergy diagnostics using the extracts of cat epithelium, birch pollen, timothy grass and their recombinant proteins as test case. The method of immunoassay on hydrogel biochip has been adapted for concurrent quantification of sIgE for 38 allergens of different classes, and principal possibility to conduct quantitative determination of sIgE and sIgG4 using biochip format has been demonstrated. (ii) A collection of the TB DNA isolates has been formed, including strains with multiple and extensive drug-resistance. Mutational profile of the strains has been established, and the association of individual TB families with resistant and sensitive forms has been shown. Phenotypic characteristics of strains resistant to bedaquiline, linezolid, perchlozone have been determined. A connection has been shown between mutations in the repressor gene Rv0678 involved in efflux system of the excretion of clofazimine and econazole, with a high level of resistance to bedaquiline. Genetic determinants of TB resistance to linezolid have been revealed, which change the structure of ribosomal peptidyl transferase center. A correlation has been suggested for the analytical microsystem based on high-sensitivity multiplex amplification with multiple “heat pulses” at the elongation stage and fluorescent labeling stage, with subsequent hybridization on the biochip. The prototype of biochip has been developed featuring an integrated personal electronic identifier containing information on the technological stages of production of the biochip and quality control procedures. Within the framework of direction “Basic mechanisms underlying human health and longevity” long-lived Drosophila lines have been obtained with enhanced activity of the genes controlling various types of DNA repair. Geroprotective role of anti-inflammatory drugs has been found for the same model. The key route of action of these drugs through a signaling cascade Pkh2 / ypk / lem3 / tat2 has been identified. Genome and transcriptome of the long-lived of the gray whale have been sequenced. Data have been obtained on the role of DNA repair in maintaining genome stability under effect of unfavorable factors and during aging. In particular the DNA repair genes Hus1, mnk, mei-9, mus210, spn-B and WRNexo have been identified, overexpression of these genes has a significant geroprotective effect. A new role of the bacterial alarmone ppGpp has been revealed as an effector of the transcription-linked DNA excision repair, reinforcing the UvrD-dependent backtracking of RNA polymerase. It has been shown that overexpression of genes katG and gshA maximizes bacterial resistance to the lethal effect of reactive oxygen species. A preparation of the key human "stress protein" Hsp70 with an optimal set of protective properties has been isolated. The effective dose of Hsp70 both for intranasal and intravenous administration has been determined. It has been found that if intranasal administration of Hsp70 is used it rapidly penetrates into the brain of mice. It has been also shown that the effect of Hsp70 depends on the age and sex of the animal, exerting a positive effect on locomotor activity and cognitive performance mostly in older animals. One-off intravenous injections of Hsp70 to male rats led to the mobilization of a number of adaptive mechanisms, increased endurance and shortened recovery time after stress.
1. Zimenkov DV, Kulagina EV, Antonova OV, Zhuravlev VY, Gryadunov DA Simultaneous drug resistance detection and genotyping of Mycobacterium tuberculosis using low-density hydrogel microarray Journal of Antimicrobial Chemotherapy, 71(6): 1520-1531 (year - 2016).
2. Garaeva A.A., Kovaleva I.E., Chumakov P.M., Evstafieva A.G. Mitochondrial dysfunction induces SESN2 gene expression through the Activating Transcription Factor 4 Cell Cycle, 15 (1), 64-71 (year - 2015).
3. Mitkevich V.A., Ilinskaya O.N., Makarov A.A. Antitumor RNases: killer’s secrets Cell Cycle, Том 14, стр. 931-932 (year - 2015).
4. Kuzmenko Y.V., Starodubova E.S., Karganova G.G., Timofeev A.V., Karpov VL Неструктурный белок 1 вируса клещевого энцефалита активирует экспрессию субъединиц иммунопротеасомы Молекулярная биология, 2, 50, 353-359 (year - 2016).
5. Matyugina E., Novikov M., Babkov D., Ozerov A., Chernousova L., Andreevskaya L., Smirnova T., Karpenko I., Chizhov A., Murthu P., Lutz L., Kochetkov S., Seley-Radtke K. L., Khandazhinskaya A.L. 5-Arylaminouracil Derivatives: New Inhibitors of Mycobacterium tuberculosis Chemical Biology and Drug Design, 86(6): 1387–1396 (year - 2015).
6. Koroleva E.P., Halperin S., Gubernatorova E.O., Macho-Fernandez E., Spencer K.M., Tumanov A.V. Citrobacter rodentium-induced colitis: A robust model to study mucosal immune responses in the gut Journal of immunological methods, 421: 61-72 (year - 2015).
7. Arefieva A.S., Smoldovskaya O.V., Tikhonov A.A., Rubina A.Y. Аллергия и аутоиммунитет: механизмы развития, молекулярная диагностика и терапия Молекулярная биология, - (year - 2016).
8. Dudeck J., Ghouse S.M., Lehmann C.H.K., Hoppe A., Schubert N., Nedospasov S.A., Dudziak D., Dudeck A. Mast-cell-derived TNF amplifies CD8 dendritic cell functionality and CD8 T cell priming Cell Reports, 13(2): 399-411 (year - 2015).
9. Alexandrova L.A., Chekhov V.O., Shmalenyuk E.R., Kochetkov S.N., El-Asrar R.A., Herdewijn P. Synthesis and evaluation of C-5 modified 2'-deoxyuridine monophosphates as inhibitors of M. tuberculosis thymidylate synthase Bioorganic and Medicinal Chemistry, 23(22): 7131-7137 (year - 2015).
10. Zimenkov D.V., Kulagina E.V., Antonova O.V., Krasnova M.A., Chernyaeva E.N., Zhuravlev V.Y., Kuz’min A.V., Popov S.A., Zasedatelev A.S., Gryadunov D.A. Evaluation of a low-density hydrogel microarray technique for mycobacterium species identification Journal of clinical microbiology, Volume 53, Number 4, pages 1103-14 (year - 2015).
11. Benezech C., Luu N.T., Walker J.A., Kruglov A.A. ... Veldhoen M., Nedospasov S.A., McKenzie A.N., Caamano J.H. Inflammation-induced formation of fat-associated lymphoid clusters Nature immunology, 16(8): 819-28 (year - 2015).
12. Kulakovskiy I.V., Vorontsov I. E., Yevshin I.S., Soboleva A.V., Kasianov A.S., Ashoor H., Ba-alawi W., Baji V.B., Medvedeva Y.A., Kolpakov F.A., Makeev V. J. HOCOMOCO: expansion and enhancement of the collection of transcription factor binding sites models Nucleic Acids Research, pii: gkv1249. [Epub ahead of print (year - 2015).
13. Mitkevich V.A., Pace C.N., Koschinski A., Makarov A.A., Ilinskaya O.N. Механизм цитотоксичности катионных мутантов РНКазыSa включает ингибирование калиевого тока через Са2+-активируемые каналы. Молекулярная биология, Том 49, №6, страницы 1041-1047 (year - 2015).
14. Danilov A., Shaposhnikov M. , Shevchenko O., Zemskaya N., Zhavoronkov A., Moskalev A. Influence of non-steroidal anti-inflammatory drugs on Drosophila melanogaster longevity Oncotarget, 6(23):19428-44 (year - 2015).
15. Proshkina E. N., Shaposhnikov M. V., Sadritdinova A. F., Kudryavtseva A. V., Moskalev A. A. Basic mechanisms of longevity: A case study of Drosophila pro-longevity genes Ageing Research Reviews, 24(Pt B):218-31 (year - 2015).
16. Shaposhnikov M., Proshkina E., Shilova L., Zhavoronkov A., Moskalev A. Lifespan and Stress Resistance in Drosophila with Overexpressed DNA Repair Genes Scientific Reports, 5:15299 (year - 2015).
17. Omelyanchuk L.V., Shaposhnikov M.V., Moskalev A.A. Drosophila nervous system as a target of aging and anti-aging interventions Frontiers in genetics, 6:89 (year - 2015).
18. Lisitsyn N.A., Chernyi A.A., Karpov V.L., Beresten S.F. Роль длинных некодирующих РНК в процессе канцерогенеза Молекулярная биология, Том 49, № 4, страницы 561–570 (year - 2015).
Annotation of the results obtained in 2016
Cytokines as a key to understanding the mechanisms and treatment of diseases The focus of the project is to study the role of cytokines and signaling cascades mediated by cytokines in the etiology of inflammatory, autoimmune and cancer diseases. A novel concept is based on the proposition that a protective as well as a pathological form of a cytokine could be produced by different cell types. Using the mouse model of collagen antibody-induced arthritis it was shown that myeloid cell-derived tumor necrosis factor (TNF) plays a pathogenic role, since mice deficient in TNF production only in myeloid cells did not develop the disease. The efficacy of pharmacological blockade of TNF by bispecific antibody MYSTI, which selectively recognizes the cytokine on macrophages, was evaluated using the TNF humanized mice and compared with the systemic TNF inhibitors (Efimov G. et al., 2016, Proc Natl Acad Sci U S A). The discovery of this specific TNF inhibitor as a potential drug for arthritis treatment was widely discussed by the scientific community including the reports in Nature Reviews Rheumatology (http://www.nature.com/nrrheum/journal/v12/n4/full/nrrheum.2016.38.html) and ScienceDaily (https://www.sciencedaily.com/releases/2016/02/160229182536.html). On the other hand, it was demonstrated that myeloid cell-derived TNF is involved in regeneration of deep skin wounds in mice. Moreover, TNF, produced by myeloid cells, performs protective function in the experimental ischemic heart disease model (Clausen B. et al., 2016, Scientific Reports). TNF, synthesized by smooth muscle cells, regulates myogenic activity of blood vessels and systemic blood pressure (Sauve M. et al., 2016, Diabetes). TNF, produced by kidney epithelial cells, increases the sensitivity to cisplatin, which could lead to the acute renal failure during chemotherapy (Zhang J. et al., 2016, J Am Soc Nephrol). The investigation of cytokines as mediators of inflammatory diseases was continued using experimental model of acute intestinal ischemia. It was shown that LTβR gene-deficient mices are more resistant to the ischemic injury of the intestinal mucosa. The novel method for the detection of the reactive oxygen species during ischemic injury of the intestine was developed. The mechanism of regulation of innate lymphoid cells during lymphotoxin-induced intestinal inflammation was revealed. It is activated through MyD88-dependent pathway in response to signals from commensal microbiota and triggers IL-22-mediated protection against intestinal inflammation. It was shown that attenuated response to T-cell receptor stimulation in Th2 cells is associated with decreased levels of Lck kinase expression. High level of IL22RA2 expression was detected in cells of human breast cancer cell line MDA-MB-468. Reporter constructs, containing necessary regulatory elements of murine and human IL22R2A genes, were designed and their activity was tested in preliminary experiments using MDA-MB-468 cells. Targets for cancer biotherapy Previously unknown details of natural anti-tumor defense mechanisms and their failures in malignant diseases were revealed. It was shown that transcription factor ATF4, that together with tumor suppressor p53 is activated in response to mitochondrial stress, regulates expression of adrenomedullin 2 gene which controls tissue reactions including metastasis (Kovaleva I. et al., 2016, Gene). The activity of homeostatic regulation genes that are regulated by p53 together with p73 is responsible for the increased survival of cancer cells with unaffected p53 gene under low-dose chemotherapy. Cancer cells with lost p53 activity become irresponsive to interferon. However, recovery of p53 function in these cells does not amend the defect of interferon system. At the same time, cervical cancer cells, where p53 is suppressed by the activity of viral genes E6 and E7, recover p53 function, interferon sensitivity and die following treatment with ribonuclease binase. The dynamics of oncolytic enteroviral adaptation revealed in cell culture and in mice, suggested that artificial destabilization of viral genome may be required for adaptational retargeting of viruses to the insensitive tumor cells. Crystals of MCT1/DENR protein complexes with 40S ribosome subunit were obtained and their 3D modeling was performed to study the oncogenic function of the proteins (Akulich K. et al., 2016, Scientific Reports). The expression profile of small serine phosphatase genes CTDSPL/1/2 was determined relevant to the stage of the clear cell renal cell carcinoma. Performed experiments allowed identifying microRNAs - potential regulators of these genes. Conditions for the detection of long non-coding RNAs (lncRNA) in serum samples of colon cancer patients were selected. Most informative lncRNAs were chosen for the analysis of cancer stem cells. A method to monitor these lncRNAs in patients was developed. Basic mechanisms underlying human health and longevity Functional annotation of genome and transcriptome of the gray whale has been completed. Molecular genetic features of the long-lived line of Drosophila melanogaster with Gclc gene overexpression were established. Lentiviral vectors have been created to suppress age-dependent activity of retrotransposon MusD. The mechanisms of anti-tumor and geroprotective properties of Abisyl drug (LLC "Initium-Farm", Russia) were revealed. Geroprotector criteria were established (Moskalev A. et al., 2016, Aging cell). “Aging Chart” - the novel online database of aging mechanisms was created (Moskalev A. et al., 2016, Nucleic Acids Res). A new DNA repair mechanism based on the interaction of RNA polymerase with protein factors UvrD and DksA in complex with ppGpp was proposed (Kamarthapu V. et al., 2016, Science). It was shown that amino acid cysteine and its derivatives play important role in protection of bacteria against oxidative stress. E.coli strain with increased production of glutathione - one of the most effective anti-oxidants was obtained. Transcriptome and histological studies of brain samples from 5XFAD (Alzheimer's disease model) mice revealed that chronic intranasal administration of human recombinant heat shock protein HSP70 has a protective "rejuvenating" effects in the various areas of the brain Significant role of Hsp70 in RNA-interference system functioning was determined (Funikov S. et al., 2016, Open Biology). A method for producing human recombinant HSP70 in the milk of transgenic mice was developed (Gurskii G. et al., 2016 Cell Stress&Chaperones). Development of drug prototypes, targeting host cell components, for the treatment of socially significant infectious diseases It was demonstrated that stabilization of the hepatitis C virus (HCV) replicon in the cell by growth factor TGF-β1 is due to the activation of noncanonical signaling pathways. The activity of matrix metalloproteinases (MMPs) and the activation of TGF-β1 is effectively inhibited by multipotent MMPs inhibitors. Previously observed antiviral effect of tubastatin A is achieved via the inhibition of histone deacetylase 6 and suppression of the enzyme expression. A new analogue of tamibarotene – TamHA was obtained. TamHA properties as a retinoic acid receptor ligand were characterized. It was shown that it has increased anti-HCV activity in comparison with the original tamibarotene. It was established that HCV replicons induce selective increase of antioxidant peroxiredoxins gene expression (Magri A. et al., 2016, Scientific reports). Structural and functional analysis of previously obtained methylene bisphosphonates identified structural elements that are necessary for the manifestation of the inhibitory activity. Eleven new compounds with improved properties were synthesized and tested (Yanvarev D. et al., 2016, Biochimie). Methoxyethylene glycol and polylysine copolymer-based nanoparticles were created in collaboration with American colleagues. The particles are able to inhibit the activity of HIV-1 reverse transcriptase and suppress viral replication in HIV-infected cells (Leporati A., et al, 2016, Nanomedicine: NBM). Obtained prototype DNA vaccines against tick-borne encephalitis virus (TBEV) were tested in cell cultures and using laboratory animals. It was shown that immunization of mice with TBEV nonstructural protein 1 (NS1) - based DNA vaccines provides partial protection from the disease. It has been found that modified variants of NS1, artificially destined for proteasomal degradation, increase life expectancy of infected animals. The optimization of structures and synthesis of fleximer analogues of 9-dezapurine bases and 5'-norcarbocyclic nucleoside analogs was performed. The molecules are potential antibacterial agents and growth inhibitors of M. tuberculosis. Molecular methods of multiplex analysis of the protein and DNA markers of socially significant diseases Biological microchips (biochips) containing sixty immobilized allergen extracts belonging to different groups were designed. A novel multiplex biochip-based immunoassay was developed to determine the concentration of specific immunoglobulins E and G4 classes in the blood serum of patients (Feyzkhanova G. et al., 2016. Clin Proteomics). The analytical performance of the method was characterized and the efficacy of the assay was demonstrated on the detection of type I hypersensitivity. The prototypes of diagnostic kits were designed. It was shown that the activity of allergens immobilized on hydrogel biochips remains intact for at least one year. An improved version of analyzer for quantitative analysis of protein biochips was designed (patent application No 2016148663 (RU), priority from 12/12/2016). The molecular mechanisms of tuberculosis causative agent resistance to bedaquiline and linezolid were revealed. Bedaquiline resistance is primarily associated with inactivation of the transcriptional repressor of mmpL5 and mmpS5 genes, whose products provide the drug efflux from the cell. For the first time mutations in the atpE gene (encodes a transmembrane ATP synthase subunit C) associated with high-level bedaquiline resistance were found in isolates from patients. Linezolid resistance is associated with mutations in genes encoding 23S rRNA and ribosomal protein L3 which alter the structure of the peptidyl transferase center of the ribosome. The prototypes of biochips with immobilized probes for the identification of genetic determinants of resistance to linezolid and bedaquiline together with a mockup of the biochip analyzer for contactless writing/reading of hybridization results from a biochip personalized electronic identifier were developed and successfully tested.
1. Akulich K.A., Andreev D.E., Terenin I.M., Smirnova V.V., Anisimova A.S., Makeeva D.S., Arkhipova V.I., Stolboushkina E.A., Garber M.B., Prokofjeva M.M., Spirin P.V., Prassolov V.S., Shatsky I.N., Dmitriev S.E. Four translation initiation pathways employed by the leaderless mRNA in eukaryotes. Scientific Reports, 6, № 37905 (year - 2016).
2. Efimov G.A., Kruglov A.A., Khlopchatnikova Z.V., Rozov F.N., Mokhonov V.V., Rose-John S., Scheller J., Gordon S., Stacey M., Drutskaya M.S., Tillib S.V., Nedospasov S.A. Cell-type-restricted anti-cytokine therapy: TNF inhibition from one pathogenic source. Proceedings of the National Academy of Sciences of the United States of America, 113(11): 3006-3011 (year - 2016).
3. Zhang J., Rudemiller N.P., Patel M.B., Wei Q., Karlovich N.S., Jeffs A.D., Wu M., Sparks M.A., Privratsky J.R., Herrera M., Gurley S.B., Nedospasov S.A., Crowley S.D. Competing Actions of Type 1 Angiotensin II Receptors Expressed on T Lymphocytes and Kidney Epithelium during Cisplatin-Induced AKI. Journal of the American Society of Nephrology, 27(8):2257-2264 (year - 2016).
4. - Онколитические вирусы Телеканал «1Медтв» Авторская программа Марины Аствацатурян «Медицина в контексте», 12.05.2016 (year - ).
5. Ivanov A.V., Valuev-Elliston V.T., Tyurina D.A., Ivanova O.N., Kochetkov S.N., Bartosch B., Isaguliants M.G. Oxidative Stress, a trigger of hepatitis C and B virus-induced carcinogenesis Oncotarget, - (year - 2016).
6. - Ученые обсудят, как продлить жизнь и победить старость Интернет-портал Наука и технологии РФ (STRF), 22.04.16 (year - ).
7. Smoldovskaya O., Feyzkhanova G., Arefieva A., Voloshin S., Ivashkina O., Reznikov Y., Rubina A. Allergen extracts and recombinant proteins: comparison of efficiency of in vitro allergy diagnostics using multiplex assay on a biological microchip Allergy, Asthma and Clinical Immunology, том 12, № 9 (year - 2016).
8. Ivanov A.V., Valuev-Elliston V.T., Ivanova O.N., Kochetkov S.N., Starodubova E.S., Bartosch B., Isaguliants M.G. Oxidative stress during HIV infection: mechanisms and consequences. Oxidative Medicine and Cellular Longevity, - (year - 2016).
9. Yanvarev D.V., Korovina A.N., Usanov N.N., Khomich O.A., Vepsäläinen J., Puljula E., Kukhanova M.K., Kochetkov S.N. Methylene bisphosphonates as the inhibitors of HIV RT phosphorolytic activity. Biochimie, 127, 153-162 (year - 2016).
10. Garbuz D.G. Регуляция экспрессии генов теплового шока в ответ на стрессовое воздействие. Молекулярная биология, - (year - 2017).
11. - Революция в онкологии. Какие вирусы могут убивать рак (Петр Чумаков) Еженедельник "Аргументы и Факты", 20.03.2016 Еженедельник "Аргументы и Факты" № 11 16/03/2016 (year - ).
12. - Cell-type-specific approach to TNF inhibition (ORIGINAL ARTICLE Efimov, G. A. et al. Cell-typerestricted anti-cytokine therapy: TNF inhibition from one pathogenic source. Proc. Natl Acad. Sci. USA http://dx.doi.org/10.1073/pnas.1520175113) Nature Reviews Rheumatology, 17.03.2016 (year - ).
13. Anisimova A.S., Rubtsov P.M., Akulich K.A., Dmitriev S.E., Frolova E., Tiulpakov A. Late Diagnosis of POMC Deficiency and in vitro evidence of residual translation from allele with c.-11C>A mutation The Journal of Clinical Endocrinology & Metabolism, - (year - 2016).
14. Leporati A., Novikov M.S., Valuev-Elliston V.T., Korolev S.P., Khandazhinskaya A.L., Kochetkov S.N., Gupta S., Goding J., Bolotin E., Gottikh M.B., Bogdanov A. Hydrophobic-core PEGylated graft copolymer-stabilized nanoparticles composed of insoluble non-nucleoside reverse transcriptase inhibitors exhibit strong anti-HIV activity. Nanomedicine: Nanotechnology, Biology, and Medicine, 12, 8, 2405-2413 (year - 2016).
15. - Флуориметрический анализатор биологических микрочипов -, 2016148663 (year - ).
16. Nosova E, Zimenkov D, Khakhalina A, Isakova A, Krylova L, Makarova M, Galkina K, Krasnova M, Safonova S, Litvinov V, Gryadunov D, Bogorodskaya E A comparison of the Sensititre MycoTB Plate, the Bactec MGIT 960, and a microarray-based molecular assay for the detection of drug resistance in clinical isolates of Mycobacterium tuberculosis in Moscow, Russia. PLoS One, PONE-D-16-28365R2 (year - 2016).
17. - Autoimmune diseases: Novel drug for arthritis sciencedaily.com, 29.02.2016 (year - ).
18. Alexandrova L.A., Efremenkova O.V., Andronova V.L., Galegov G.A., Solyev P.N., Karpenko I.L., Kochetkov S.N. 5-(4-Алкил-1,2,3-триазол-1-ил)метильные производные 2’-дезоксиуридина – ингибиторы роста бактерий и вирусов. Биоорганическая химия, 42, 6, 746-754 (year - 2016).
19. - Результаты первого года Комплексной научной программы, финансируемой РНФ и выполняемой ИМБ РАН Пресс-служба Российского научного фонда, о результатах первого года Комплексной научной программы, финансируемой РНФ и выполняемой ИМБ РАН., Фильм демонстрировался на заседании представителей попечительского и экспертного советов РНФ в Нижнем Новгороде 24-25 февраля 2016 г. Результаты работы сотрудников ИМБ РАН получили высокую оценку со стороны руководства Фонда (year - ).
20. Kamarthapu V., Epshtein V., Benjamin B., Proshkin S., Mironov A., Cashel M., Nudler E. ppGpp couples transcription to DNA repair in E. coli. Science, 352, 6288, 993-996 (year - 2016).
21. Gurskiy Y.G., Garbuz D., Soshnikova N.V., Krasnov A.N., Deikin A., Lazarev V.F., Sverchinskyi D., Margulis B.A., Zatsepina O.G., Karpov V.L., Belzheralskaya S., Feoktistova E, Georgieva S.G., Evgen’ev M.B. The development of modified human Hsp70 and its production in the milk of transgenic mice. Cell Stress and Chaperones, 21, 1055-1064 (year - 2016).
22. - Выпуск новостей Телеканал Россия 1, 15.03.16 (year - ).
23. Gubernatorova E.O., Perez-Chanona E., Koroleva E.P., Jobin C., Tumanov A.V. Murine Model of Intestinal Ischemia-reperfusion Injury. Journal of Visualized Experiments, (111), e53881e53881 (year - 2016).
24. Zatsepina O.G., Przhiboro A.A., Yushenova I.A., Shilova V., Zelentsova E.S., Shostak N.G., Evgen'ev M.B., Garbuz D.G. A Drosophila heat shock response represents an exception rather than a rule amongst Diptera species. Insect Molecular Biology, 25, 4, 431-449 (year - 2016).
25. Clausen B.H., Degn M., Sivasaravanaparan M., Fogtmann T., Andersen M.G., Trojanowsky M.D., Gao H., Hvidsten S., Baun C., Deierborg T., Finsen B., Kristensen B.W., Bak S.T., Meyer M., Lee J., Nedospasov S.A., Brambilla R., Lambertsen K.L. Conditional ablation of myeloid TNF increases lesion volume after experimental stroke in mice, possibly via altered ERK1/2 signaling. Scientific Reports, 6:29291 (year - 2016).
26. Andreeva N.V., Zatsepina O.G., Garbuz D.G., Evgen'ev M.B., Belyavsky A.V. Recombinant HSP70 and mild heat shock stimulate growth of aged mesenchymal stem cells. Cell Stress and Chaperones, 21, 4, 727-733 (year - 2016).
27. - ГЗК П-53 - страж генома, регулирующий судьбу наиважнейших процессов в клетках, от которого зависит здоровье или нездоровье человека Телеканал Культура Программа Черные дыры. Белые пятна, 7.04.2016 (year - ).
28. - Двурушничество как точная наука_ Сайт РНФ, 15.03.16 (year - ).
29. Proshkin S.A., Mironov A.S. Остановленная РНК-полимераза – мишень фактора Mfd. Молекулярная биология, 50, 2, 332-335 (year - 2016).
30. - Рак уже не приговор? Какие методы диагностики и лечения существуют Еженедельник "Аргументы и Факты", 05.06.2016, Журнал «АиФ Про Здоровье» №6 (104), июнь 2016 (year - ).
31. Kuzmenko Y.V., Smirnova O.A., Ivanov A.V., Starodubova E.S., Karpov V.L. Nonstructural protein 1 of tick-borne encephalitis virus induces oxidative stress and activates antioxidant defense Nrf2/ARE pathway. Intervirology, 59, 111-117 (year - 2016).
32. - Онколитические вирусы (повтор) Телеканал «1Медтв» Авторская программа Марины Аствацатурян «Медицина в контексте», 14.06.2016 (year - ).
33. - Революция в онкологии. Какие вирусы могут убивать рак Независимое интернет-издание «Новостная лента», 20.03.2016 (year - ).
34. - Recombinant allergens show similar diagnostic accuracy to natural extracts (по материалам статьи Smoldovskaya O, et al. Allergy Asthma Clin Immunol. 2016;doi:10.1186/s13223-016-0117-1) Healio.com, 29.04.2016 (year - ).
35. Beniaminov A.D., Krasnov G.C., Dmitriev A.A., Puzanov G.A., Snopok B.A., Senchenko V.N., Kashuba V.I. Взаимодействие двух супрессоров опухолевого роста фосфатазы CTDSPL и белка Rb Молекулярная биология, том 50, № 3, 504-508 (year - 2016).
36. Yanvarev D.V., Korovina A.N., Usanov N.N., Khomich O.A., Vepsäläinen J., Puljula E., Kukhanova M.K., Kochetkov S.N. Data on synthesis of methylene bisphosphonates and screening of their inhibitory activity towards HIV reverse transcriptase. Data in Brief, 8, 1157-1167 (year - 2016).
37. Antonova A.V., Gryadunov D.A., Zimenkov D.V. Молекулярные механизмы лекарственной толерантности Mycobacterium tuberculosis Молекулярная биология, - (year - 2017).
38. Yurinskaya M.M., Kochetkova O.Yu., Shabarchina L., Antonova O.Yu., Suslikov A.V., Evgen’ev M.B., Vinokurov M.G. Encapsulated Hsp70 decreases endotoxin-induced production of ROS and TNFα in human phagocytes. Cell Stress and Chaperones, - (year - 2016).
39. Kovaleva I.E., Garaeva A.A., Chumakov P.M., Evstafieva A.G. Intermedin/adrenomedullin 2 is a stress-inducible gene controlled by activating transcription factor 4 Gene, Том 590, № 1, 177-185 (year - 2016).
40. Feyzkhanova G., Voloshin S., Smoldovskaya O., Arefieva A., Filippova M., Barsky V., Pavlushkina L., Butvilolvskaya V., Tikhonov A., Reznikov Y., Rubina A. Development of microarray-based method for allergen-specific IgE and IgG4 detection Clinical Proteomics, - (year - 2017).
41. Moskalev A., Shaposhnikov M., Proshkina E., Krasnov G., Belyi A., Fedintsev A., Zhikrivetskaya S., Sadritdinova A., Snezhkina A., Zhavoronkov A., Guvatova Z., Kudryavtseva A. The influence of pro-longevity gene Gclc overexpression on the age-dependent changes in Drosophila transcriptome and biological functions BMC Genomics, - (year - 2016).
42. - Двурушничество как точная наука Интернет-портал Сноб, 15.03.16 (year - ).
43. - Ученые создали прототип лекарства против артрита и псориаза Интернет-портал Наука и технологии РФ (STRF), 1.03.2016 (year - ).
44. - Ученые создали прототип лекарства против артрита, псориаза Интернет-портал Газета.ru, 1.03.2016 (year - ).
45. Kudryavtseva A., Krasnov G., Lipatova A., Alexeev B., Maganova F., Shaposhnikov M., Fedorova M., Snezhkina A., Moskalev A. Effects of Abies sibirica terpenes on cancer- and aging-associated pathways in human cells. Oncotarget, - (year - 2016).
46. Moskalev A., Zhikrivetskaya S., Shaposhnikov M., Dobrovolskaya E., Gurinovich R., Kuryan O., Pashuk A., Jellen L.C., Aliper A., Peregudov A., Zhavoronkov A. Aging Chart: a community resource for rapid exploratory pathway analysis of age-related processes. Nucleic Acids Research, 44, D894-D899 (year - 2016).
47. Funikov S.Yu., Ryazansky S.S., Kanapin A.A., Logacheva M.D., Penin A.A., Snezhkina A.V., Shilova V.Yu., Garbuz D.G., Evgen’ev M.B., Zatsepina O.G. The effect of severe heat shock on microRNA expression pattern in Drosophila melanogaster. Open Biology, 6 (year - 2016).
48. Sauvé M., Hui S.K., Dinh D.D., Foltz W.D., Momen A., Nedospasov S.A., Offermanns S., Husain M., Kroetsch J.T., Lidington D., Bolz S.S. Tumor Necrosis Factor/Sphingosine-1-Phosphate Signaling Augments Resistance Artery Myogenic Tone in Diabetes. Diabetes, 65(7):1916-1928 (year - 2016).
49. Magri A., Ozerov A.A., Tunitskaya V.L., Valuev-Elliston V.T., Wahid A., Pirisi M., Simmonds P., Ivanov AV, Novikov M.S., Patel A.H. Exploration of acetanilide derivatives of 1-(ω-phenoxyalkyl) uracils as novel inhibitors of Hepatitis C Virus replication. Scientific reports, 6, 29487 (year - 2016).
50. Moskalev A, Chernyagina E, Tsvetkov V, Fedintsev A, Shaposhnikov M, Krut'ko V, Zhavoronkov A, Kennedy BK. Developing criteria for evaluation of geroprotectors as a key stage toward translation to the clinic. Aging Cell, 15, 3, 407-415 (year - 2016).
51. Kozlov M.V., Kleymenova A.A., Konduktorov K.A., Malikova A.Z., Kamarova K.A., Novikov R.A., Kochetkov S.N. Синтез (Z)-N-гидрокси-3-метокси-3-фенилакриламида – нового селективного ингибитора репликации вируса гепатита С. Биоорганическая химия, 42, 2, 191-197 (year - 2016).
52. Vorontsov I.E., Khimulya G., Lukianova E.N., Nikolaeva D.D., Eliseeva I.A., Kulakovskiy I.V., Makeev V.J. Negative selection maintains transcription factor binding motifs in human cancer BMC Genomics, 17, 395, 263-276 (year - 2016).
Annotation of the results obtained in 2017
Within the scope of the project entitled "Cytokines - the key to understanding the mechanisms and therapy of diseases” are the cytokines and the signaling cascades which they mediate as possible targets for the therapy of autoimmune, inflammatory diseases and cancer. New properties of bispecific human TNF inhibitors have been established, an effective dose of the inhibitor has been established, associated with complete or partial protection of hTNF KI mice in the experimental model of arthritis. The effectiveness of the use of the bispecific TNF inhibitor in the experimental autoimmune encephalomyelitis has also been tested. New functions of cytokines have been established, in particular, the involvement of TNF produced by myeloid cells in the activity of hair follicles during skin wound regeneration and the role of TNF in the regulation of muscle activity and systemic blood pressure, not directly related to the immune system. Studies of mice with genetic defects in cytokines and cytokine signaling cascades have shown that TLR4 expressed by myeloid cells, unlike TNF, is not critical for LPS/D-Gal-induced hepatotoxicity. Finally, mice with TNF knockout in macrophages and in T cells are resistant to the development of ischemic intestinal damage. "Targets for cancer biotherapy" Several mechanisms involved in tumor cell survival under cytotoxic therapy were identified. Induction of p53-dependent genes involved in autophagy was revealed as a mechanism that stands behind relative resistance of tumor cells to chemotherapy. Three new genes regulated by transcription factor ATF4 and involved in overcoming ER stress consequences were identified. A mechanism of mitochondria function regulation by the hsa-miR4485 microRNA which is involved in tumor suppression has been established. Interferon response signaling pathway defect, associated with decreased level of the STAT2 protein and causing the acquisition of sensitivity to oncolytic viruses was revealed in glioblastoma cell line. Several strains of oncolytic Coxsackie virus B6 capable to efficiently replicate and possessing extended tropism with respect to the diversity of human tumor cells were obtained through bioselection on cells previously insensitive to the virus. It has been established that apoptogenic effect of cytotoxic ribonuclease binase on tumor cells is due to alterations in the redox balance, associated with changes in the level of glutathionylation of proteins involved in the differentiation and control of proliferative signaling pathways. By this mechanism binase increases sensitivity to anti-cancer drugs. Cell lines with silenced MCTS1, DENR and eIF2D genes were obtained. Transcriptome analysis of these cells revealed significant changes in the expression of genes associated with translational regulation and with the regulation of transcription of genes involved in malignant transformation. In addition, MCTS1, DENR and eIF2D genes knockout cell lines were obtained using CRISPR/Cas9 technology. The lines will be used to create a test system for the search for inhibitors of protein complexes based on fluorescent complementation for the analysis of complexation of proteins in extracts. The structure of the MCTS1/DENR heterodimer with human ribosome was determined.This allowed us to propose a model describing the role of these translation factors in the re-initiation of translation and ribosome recycling. A long non-coding RNA LINC00973, a new biomarker for the prognosis of colon cancer was discovered. It represents a promising target for chemotherapy of the most common types of tumors. "Basic mechanisms underlying human health and longevity" 1.7.1. Basic classification of geroprotectors providing homeostasis of an organism has been created. The compounds include: the ones eliminating the consequences of homeostasis disturbances; enhancing the ability of endogenous homeostatic systems; neutralizing the damaging influence of environmental factors and, hence, leading to homeostasis failure; inhibiting excessive homeostatic reactions, leading to further homeostasis failure. The major parameters of age dynamics of stress resistance, fertility, neuro muscle activity, circadian rythms and trnscrtiptomic changes after introduction of potential geroprotector fuvoxanthine have been investigated. Geroprotective effects of chronic action of flavonoids quarcetine and (-)-epikatkhin were studied in detail. Analytical mathematic model for evaluation of biological age of humans basing on 89 parameters of aging has been generated. The differences in reaction of grey whale to DNA damage and hypoxia with other long-lived organisms have been determined. New components of Mg homeostasis in yeast that are homologous to the correspondent human genes were described. 1.7.2. The failure of DNA reparation exploring Base Excision Repair (BER) observed in E.coli strain defective by ppGpp synthesis, as well as in the strain with the deleted gene dksA, enables to postulate two independent pathways of BER, one of them coupled with transcription (TCR). In the course of our studies of interaction between cystein metabolism, gene mstA activity and generation of reactive oxygene species (ROS) we revealed the mechanisms underlying protective effects H2S from oxidative stress. It was demonstrated that H2S sequestered free Fe in Fenton reaction and, hence, prevents the generation of hydroxyl radical toxic for cells. 1.7.3. Complex investigation of human recombinant human Hsp70 has been performed and the optimal storage conditions have been determined. Mechanisms of Hsp70 proteolysis in culture cells as well as in the brain of experimental animals after intranasal administration have been described in detail. The active doze of Hsp70 for intranasal administration and in cell cultures has been estimated. It was shown that recombinant Hsp70 after chronic intranasal injections does not induce allergic reaction or exhibit immunotoxic properties, which make this compound a promising therapeutic drug to treat various neuropathologies in humans. "Development of drug prototypes, targeting host cell components, for the treatment of socially significant infectious diseases" It has been shown that the reproduction of hepatitis C virus replicon in hepatocyte culture is supported by class I, IIa and IIb histone deacetylases. The antiviral effect of previously synthesized 4-pyridoxy derivatives of cinnamic hydroxamic acid is associated with the impact on several classes of histone acetylases and degradation of viral enzymes NS3 and NS5B. We obtained plasmids and isolated enzymes necessary to create cellular system, which could be used to analyze the influence of low molecular weight inhibitors on the biosynthesis of terpenoids during HIV infection. Prototype DNA vaccines against tick-borne encephalitis virus, based on its NS1 protein, were analyzed and compared in cell culture and in mouse model. A DNA vaccine against the Lassa virus was proposed. A prototype vaccine against rabies virus was tested. Novel biologically active compounds demonstrating anti-HIV and anti-tuberculosis activities simultaneously in combination with low cytotoxicity were synthesized. The growth of parasites Trypanosoma brucei and Leishmania Mexicana was inhibited by obtained 5'-norcarbocyclic pyrimidine nucleosides. "Molecular methods of multiplex analysis of the protein and DNA markers of socially significant diseases" The study of sIgE and sIgG4 profiles from the patients with different allergic diseases showed the increase of frequency of detected levels of sIgE and sIgG4 to inhaled allergens with the growth of age of patients. It has been also shown that sIgG4-response to inhaled allergens develops primarily when the sIgE levels to these allergens are increased. This explains a more frequent incidence of increased sIgG4 levels for allergic rhinitis and bronchial asthma as compared with atopic dermatitis. To investigate the contribution of allergen-specific immunoglobulins into the emergence of allergic reactions, the biochip-based assay for multiplex analysis of IgG, IgA, and IgM was developed. Laboratory trials of prototype of the ALLERGO-BIOCHIP diagnostic kit for the determination of sIgE levels to 44 allergens belonging to different groups were successful. Results of investigation of M. tuberculosis isolates that were resistant to bedaquiline and linezolid indicate the necessity of the analysis of mmpR, atpE, rrl, rplC loci, which are directly involved into resistance acquisition, and also other gene markers affecting the phenotype of isolates, for which the drug tolerance is achieved by alteration to metabolically inactive form and persistence or by activation of the basal resistance mechanisms to newest anti-tuberculosis drugs. The latter schemes include determinants in regulons participated into cell metabolism and cell wall synthesis, activation of transport proteins, and control of transcription of genes involved in lipid metabolism system. The prototype of hydrogel biochip-based TB-Biochip-IV diagnostic kit was developed and successfully tested for the primary screening of mutations in Rv0678, atpE, rrl, rplC loci.
1. Svyatchenko V.A., Ternovoy V.A., Kiselev N.N., Demina A.V., Loktev V.B., Netesov S.V., Chumakov P.M. Bioselection of Coxsackievirus B6 strain variants with altered tropism to human cancer cell lines Archives of Virology, Volume: 162 Issue: 11 Pages: 3355-3362 (year - 2017).
2. Evgen’ev M.B., Krasnov G.S., Nesterova I.V., Garbuz D.G., Karpov V.L., Morozov A.V., Snezhkina A.V., Samokhin A.N., Sergeev A., Bobkova N.V. Molecular mechanisms underlying neuroprotective effect of eHsp70 in transgenic 5XFAD mice Journal of Alzheimer’s disease, Volume: 59 Issue: 4 Pages: 1415-1426 (year - 2017).
3. Mironov A., Seregina T., Nagornykh M., Luhachack L.G., Korolkova N., Lopes L.E., Kotova V., Zavilgelsky G., Shakulov R., Shatalin K., Nudler E. Mechanism of H2S-mediated protection against oxidative stress in Escherichia coli Proceedings of the National Academy of Sciences of the United States of America, Volume: 114 Issue: 23 Pages: 6022-6027 (year - 2017).
4. - Биочипы — высокие технологии в медицинской диагностике Интернет-портал "Элементы.ру", 9.10.2017 (year - ).
5. Kuzmenko Y.V., Starodubova E.S., Shevtsova A.S., Chernokhaeva L.L., Latanova A.A., Preobrazhenskaia O.V., Timofeev A.V., Karganova G.G., Karpov V.L. Intracellular degradation and localization of NS1 of tick-borne encephalitis virus affect its protective properties Journal of General Virology, 98, 50-55 (year - 2017).
6. - Русские ученые: экстракт пихты удерживает юность и лечит рак Познавательный туристический портал «GURSESINTOUR.com», 28.02.2017 (year - ).
7. Moskalev A., Chernyagina E., Kudryavtseva A., Shaposhnikov M. Geroprotectors: A Unified Concept and Screening Approaches Aging and Disease, Volume: 8 Issue: 3 Pages: 354-363 (year - 2017).
8. Sripada L., Singh K., Lipatova A.V., Singh A., Prajapati P., Tomar D., Bhatelia K., Roy M., Singh R., Godbole M.M., Chumakov P.M., Singh R. hsa-miR-4485 regulates mitochondrial functions and inhibits the tumorigenicity of breast cancer cells Journal of Molecular Medicine, 95(6), 641-651 (year - 2017).
9. - Сюжет Вестей о разработках ИМБ РАН, посвященных диагностике аллергии «Телеканал Россия 1», 12.04.2017, «Вести» Эфир в 20:00 (year - ).
10. Savvateeva-Popova E.V., Zhuravlev A.V., Brazda V., Zakharov G.A., Kaminskaya A.N., Medvedeva A.V., Nikitina E.A., Tokmatcheva E.V., Dolgaya J.F., Kulikova D.A., Zatsepina O.G., Funikov S.Y., Ryazansky S.S., Evgen'ev M.B. Drosophila Model for the Analysis of Genesis of LIM-kinase 1-Dependent Williams-Beuren Syndrome Cognitive Phenotypes: INDELs, Transposable Elements of the Tc1/Mariner Superfamily and MicroRNAs Frontiers in Genetics, Volume: 8 Article Number: 123 (year - 2017).
11. - В пихте нашли вещества с противораковым эффектом Научно-популярный мультимедийный портал, «Чердак», 22.11.2017 (year - ).
12. Gaman M.S., Matyugina E.S., Novikov M.S., Babkov D.A., Solyev P.N., Kochetkov S.N., Khandazhinskaya A.L. New benzophenone phosphonate derivatives Mendeleev Communications, 27, 346–348 (year - 2017).
13. - Ученые обнаружили, что экстракт пихты обладает противораковыми свойствами Сайт информационного агентства «INNOV.RU» (Российский бизнес on-line), 27.02.2017 (year - ).
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