Laboratory of Molecular Endocrinology

The laboratory was founded in 1982. The main areas of scientific research are the study of the processes of hormone reception, cytokines and growth factors, and their intracellular signaling. Elucidation of the molecular mechanisms of regulation of tissue growth, migration, proliferation and differentiation of cells. Studies of the components of the fibrinolytic system and navigation receptors in the processes of vascular and nerve growth. It was found that damage to the vessel leads to the expression of urokinase and its receptor in vascular cells, which are key participants in the migration of blood cells and blood vessels into the lumen of the vessel. For the first time, the laboratory staff determined the tertiary structure of urokinase and its complex with the urokinase receptor. Biosensory technologies and intravital microscopy of single cells were used. The obtained results allowed us to hypothesize that hydrogen peroxide acts as a secondary mediator and mediates the activating effect of growth factors, cytokines and urokinase on cell migration and proliferation. The laboratory actively uses approaches related to the CRISPR/Cas9 genome editing technology. In particular, various cell lines and animals created on the basis of this method, which can be further used to identify new potential therapeutic targets or to screen potential drugs for the treatment of various heart and vascular pathologies. Current research in the laboratory is aimed at elucidating the molecular mechanisms of regulation of directed vascular and nerve growth involving the urokinase system. The role of the urokinase system in the processes mediating wound healing and fibrosis, as well as in the epithelial-mesenchymal transition, is being investigated. A large scientific reserve has been accumulated on the role of urokinase and its receptor in the morphogenesis of the brain and in the restoration of damaged vessels and nerves. The binding of urokinase to the receptor is being studied, leading to the launch of signaling cascades inside the cell, leading to the restructuring of the cytoskeleton, redistribution of adhesive contacts, stimulation of adhesion, migration and proliferation of cells. For the first time, data have been obtained on the navigational role of the urokinase system in regulating the growth trajectory of blood vessels and nerves. The most significant research results of recent years have studied the mechanisms of vascular cell tolerance to the effects of hormones and pharmacological drugs and shown the role of hypoxia in these processes. The interaction of urokinase with the extracellular matrix protein fibulin-5 has been established. Fibulin-5 has been shown to affect such integrin-dependent effects of urokinase as cell proliferation and migration, and intracellular signaling. A new urokinase signaling pathway has been identified that regulates gene expression, mediated by rapid nucleolin-dependent translocation of urokinase into the nucleus, and is responsible for the phenotypic transformation of fibroblasts into myofibroblasts. Based on NMR analysis data, a new mechanism of interaction of urokinase with the urokinase receptor is described, according to which β-layers in the growth domain are not a necessary element for interaction with the urokinase receptor. Laboratory staff isolated and characterized mesenchymal stem cells (MSCs) of human adipose tissue. The ability of these cells to stimulate tissue regeneration and induce the growth of blood vessels and nerves has been discovered. The mechanisms of such stimulation have been identified, and it has been established that the stimulating effect of MSCs is due to both soluble secretion products (growth factors, cytokines, and chemokines) and the generation of microvesicles. The effect of inflammation and hypoxia on the functional activity of these cells was assessed. It has been established that the migration rate of mesenchymal cells is controlled by a receptor-dependent mechanism, including activation of NADPH oxidases (NOX), formation of reactive oxygen species, and accumulation of hydrogen peroxide inside cells. A new transcription factor Prep1 and a mechanism for regulating the activity of PPARy, a master regulator of adipogenesis and lipid metabolism, are described. It has been shown that Prep1 belongs to a family of homeobox-containing factors that are critically important for cell differentiation, including in the adipocytic direction. There is evidence that Prep1 can regulate the sensitivity of cells to the action of insulin. It has been established that the urokinase system provides regeneration of peripheral nerves. Using genetically modified mouse lines lacking the urokinase (uPA-/-) gene and the urokinase receptor (uPAR-/-), the mechanism of the urokinase receptor's influence on post-traumatic nerve regeneration was established: it was shown that the lack of uPAR expression in regenerating axons leads to a delay in functional nerve repair, rapid axonal degeneration, and slowing of Schwann cell proliferation. For the first time, it has been shown that uPAR has its own effects in regeneration, which do not depend on the presence of a proteolytically active urokinase in the system. Investigating the molecular mechanisms of this action, it was found that uPAR stimulates the assembly of the signaling system with the participation of integrins α5ß1, an important participant in cell adhesion. It has been shown that its interaction with receptor tyrosine kinases, the epidermal growth factor EGFR receptor and the neurotrophin receptor TrkC, plays an important role in the mechanism of directed nerve growth involving uPAR. Activation of intracellular signaling involving uPAR and EGFR/TrkC has also been found to mediate intracellular signaling regulating neuron survival and differentiation. Since the urokinase system plays an important role in carcinogenesis and metastasis, the use of approaches to reduce its activity or knock out is a promising approach for the creation of anticancer drugs. Using the CRISPR/cas9 genome editing system to delete the uPAR gene, we have shown that knockout of uPAR in neuroblastoma cells leads to a significant decrease in their proliferation, DNA degradation, and caspase-dependent cell death. List of main publications: 2018. Involvement of the Urokinase Receptor and Its Endogenous Ligands in the Development of the Brain and the Formation of Cognitive Functions. Semina E.V., Rubina K.A., Stepanova V.V., Tkachuk V.A. in the Journal of Neuroscience and Behavioral Physiology, Kluwer Academic/Plenum Publishers (United States), vol. 48, No. 1, pp. 16-27 2018. Mechanisms of regulation of the directed growth of vessels and nerves by the fibrinolytic system components and GPI-anchored navigation receptors. Rubina K.A., Semina E.A., Balatskaya M.N., Plekhanova O.S., Tkachuk V.A. in the journal of Neuroscience and Behavioral Physiology, Kluwer Academic/Plenum Publishers (United States), volume 104, No. 9, pp. 1001-1026 2018. CRISPR/Cas9 nickase mediated targeting of urokinase receptor gene inhibits neuroblastoma cell proliferation. Rysenkova Karina D., Semina Ekaterina V., Karagyaur Maxim N., Shmakova Anna A., Dyikanov Daniyar T., Vasiluev Petr A., Rubtsov Yury P., Rubina Kseniya A., Tkachuk Vsevolod A. in Oncotarget magazine, Impact Journals Publishing house (Albany, N.Y., United States), volume 9, No. 50, pp. 29414-29430, 2018. Mechanisms of regulation of directed nerve and vascular growth by components of the fibrinolytic system and GPI-anchored navigation receptors Rubina K.A., Semina E.A., Balatskaya M.N., Plekhanova O.S., Tkachuk V.A. in the Sechenov Russian Journal of Physiology, Nauka Publishing House (St. Petersburg), volume 104, No. 9, pp. 1001-1026, 2018. The use of CRISPR/Cas9 genome editing technology to suppress the expression of the urokinase receptor gene in neuroblastoma cells. Rysenkova K.D., Semina E.V., Karagyaur M.N., Shmakova A.A., Dyykanov D.T., Rubina K.A., Tkachuk V.A. in the Journal Technologies of Living Systems, Radiotechnika Publishing House (Moscow), volume 15, No. 1, pp. 10-19 2016. Urokinase and urokinase receptor participate in regulation of neuronal migration, axon growth and branching. Semina E.V., Rubina K.A., Sysoeva Veronika, Rysenkova Karina, Klimovich Polina, Plekhanova Olga, Tkachuk Vsevolod in the European Journal of Cell Biology, Elsevier BV publishing house (Netherlands), volume 95, No. 9, pp. 295-310 2017. Navigation molecules and chemokines in the processes of vascular growth and remodeling. Rubina K.A., Semina E.V., Tkachuk V.A. in the journal Journal of Evolutionary Biochemistry and Physiology, Nauka Publishing House. St. Petersburg Publishing House (St. Petersburg), No. 5, pp. 313-327 2016. Participation of Urokinase in Vascular Cell Migration and in Regulation of Growth and Branching of Capillaries. Semina E.V., Rubina K.A., Sysoeva V.Yu, Makarevich P.I., Parfyonova Ye V., Tkachuk V.A. in the journal Cell and Tissue Biology, publisher Maik Nauka/Interperiodica Publishing (Russian Federation), volume 10, No. 1, pp. 37-46 2016. Three-Dimensional Model of Biomatrix as a Method of Studying Blood Vessels and Nerve Growth in Tissue Engineering Structures. Semina E.V., Rubina K.A., Sysoeva V.Yu, Stepanova V.V., Tkachuk V.A. in the Moscow University Chemistry Bulletin, Allerton Press Inc. (United States), vol. 71, No. 3, pp. 172-177 2015. The involvement of the urokinase system in the migration of vascular cells and in the regulation of capillary growth and branching. Semina E.V., Rubina K.A., Sysoeva V.Yu., Makarevich P.I., Parfenova E.V., Tkachuk V.A. in the journal Cytology, Nauka Publishing House (St. Petersburg), volume 57, No. 10, pp. 689-698 2013. Prospects for the creation of antitumor drugs aimed at the urokinase-type plasminogen activator system. Beloglazova I.B., Hakobyan Zh.A., Karagyaur M.N., Plekhanova O.S., Semina E.V., Stambolsky D.V. in the Journal Technologies of Living Systems, Radiotechnika Publishing House (Moscow), volume 10, No. 1, pp. 3-19. 2018. The niche of the stem cell. Nimiritsky P.P., Sagaradze G.D., Efimenko A.Yu., Makarevich P.I., Tkachuk V.A. in the journal Cytology, Nauka Publishing House (St. Petersburg), volume 60, No. 8, pp. 575-586