Vol 12, No 4 (2017)

Full Issue


Exosomes: from biology to clinics

Samoylova E.M., Kalsin V.A., Bespalova V.A., Devichensky V.M., Baklaushev V.P.


Exosomes are extracellular vesicles with the diameter of 30-120 nm, originating from early endosomes. Exosomes have been actively studied in the last decade, and a great amount of data has appeared on their nature and role in the intercellular transport and signaling both in the normal and pathological conditions. A particular interest to exosomes in the clinical practice emerged after the separation of their circulating fraction from the blood and the study of tumor genetic markers in them became possible (so called “liquid biopsy”). The objective of this review is to familiarize clinical specialists with the fundamentals of exosomes' biology and physiology and with the main achievements on their practical application in the medicine, as a natural drug delivery system, as well as for high-precision, early non-invasive differential diagnostics of diseases.
Genes & Cells. 2017;12(4):7-19
pages 7-19 views

Role of the different signal transduction systems on intervertebral disk degeneration

Byvaltsev V.A., Garashchenko N.E., Stepanov I.A., Aldiyarova N.T.


The review systematized the modern data reported about signaling system role in the intervertebral disk degeneration, that is one of the main reasons for back pain and patients' disability as a result. We represented current data revealing molecular-cell mechanisms of disk degeneration. The experimental data analysis of the cytokines and biologic active molecules influencing on the intervertebral disk degenerative diseases development is presented. We denoted actual, still remaining unsolved questions concerning treatment of the diseases that required further experimental studies with opportunity for clinical translation.
Genes & Cells. 2017;12(4):20-25
pages 20-25 views

Expression pattern and alternative splicing of HTT gene in human tissues

Malakhova A.A., Elisaphenko E.A.


The HTT gene (Huntingtin, IT-15) was described in 1993 as highly expressed in various parts of the brain and other human and rodent tissues. The interest to this gene is due to the fact that the expansion of trinucleotide repeats in the first exon leads to the Huntington's disease. However, the causes of selective death of striata neurons in the course of the disease development are still unknown. Studying the HTT expression pattern in different tissues allows us to understand the role of HTT isoforms in different human tissues and organs. We studied the expression and alternative splicing of HTT in different parts of the brain and other human tissues in healthy people and Huntington's disease patients. No aberrant HTT forms were found in striatal neurons. This confirms the important role of the HTT gene for this type of neurons.
Genes & Cells. 2017;12(4):26-32
pages 26-32 views

Expression dynamics of the serotonergic system components in granulosa cells of the developing ovarian follicle and after luteinization

Nikishin D.A., Alyoshina N.M., Semenova M.L., Shmukler Y.B.


Regulation of the growth and development of female sex cells is one of the conservative functions of serotonin, but their specific mechanisms remain undisclosed yet. Molecular-genetic study of the presence of all components of the serotonergic system and dynamics of their expression in granulosa cells at different stages of folliculogenesis and in corpus luteum was carried out by reverse transcription with PCR and real-time PCR. Transcripts of enzymes tryptophan hydroxylase Tph1, aromatic amino acids decarboxylase Ddc and monoamine oxidase Maoa, membrane serotonin transporter Sert and vesicular monoamine transporter Vmat2, as well as serotonin receptors Htrlb, Htrld, Htr2a, Htr2b, Htr5b and Htr7 are detected in probes investigated. A quantitative study revealed that the expression of Vmat2, Htrlb, and Htr7 genes is greater in the early stages of folliculogenesis, whereas the expression of the Vmatl gene grows by the late stages of follicular development. The relative quantity of the mRNA of the Ddc, Maoa, Vmatl and Htrlb genes increases during luteinization, while the expression of the Vmat2 and Htr7 genes, on the contrary, decreases. The presence of expression of key components of the serotonergic system and its dynamics allow as to suggest that they are involved in serotonergic regulation of folliculogenesis, as well as luteinization.
Genes & Cells. 2017;12(4):33-38
pages 33-38 views

Biological activity comparative evaluation of the gene-activated bone substitutes made of octacalcium phosphate and plasmid DNA carrying VEGF and SDF genes: part 2 - in vivo

Bozo I.Y., Rozhkov S.I., Komlev V.S., Volozhin G.A., Eremin I.I., Smirnov I.V., Savva O.V., Isaev A.A., Popov V.K., Drobyshev A.Y., Deev R.V.


Bone substitutes with osteoinductive and (or) osteogenic capacities are highly needed in clinical practice for treatment of patients with skeletal bone pathology. Gene-activated bone substitutes consisting of a scaffold and gene constructs belong to such materials. In this study, we made two proto-types of gene-activated bone substitutes based on octacalcium phosphate (OCP) granules and two variants of plasmid DNA - the system delivering single gene encoding vascular endothelial growth factor A-165 (VEGFA-165, pl-VEGFA) and the other plasmid carrying simultaneously VEGFA and gene of stromal-derived factor 1a (pl-VEGFA-SDF). All the materials were implanted to rabbits into the full-thickness parietal bone defects with diameter 10 mm, OCP without plasmid DNA we used as a control. Both gene-activated materials showed pronounced osteoinduction providing new bone formation in the central part of the defects and complete parietal bone repair by 90 days after surgery. In addition, we found newly formed bone level to be higher in pl-VEGFA-SDF group comparing with pl-VEGFA (p<0.05), while there were the opposite results at 60 days' time point (p<0.05). Thus, gene-activated bone substitutes we developed had osteoinductive effect, dynamic and features of which were different and depended on biological action of the factors delivered by transgenes.
Genes & Cells. 2017;12(4):39-46
pages 39-46 views

Modification of biodegradable fibrous scaffolds with Epidermal Growth Factor by emulsion electrospinning for promotion of epithelial cells proliferation

Tenchurin T.H., Lyundup A.V., Demchenko A.G., Krasheninnikov M.E., Balyasin M.V., Klabukov I.D., Shepelev A.D., Mamagulashvili V.G., Orehov A.S., Chvalun S.N., Dyuzheva T.G.


Supporting of a physiologically relevant cellular micro-environment is currently a grand challenge in the design of tissue-engineering grafts based on biocompatible and biodegradable polymeric materials. The aim of this research was to develop a new technique of a fibrous polycaprolactone-based scaffold modification with epidermal growth factor (EGF) and assessment of its effect on scaffold properties and proliferative activity of epithelial cells in vitro. Fibrous scaffolds from EGF-functionalized polycaprolactone has received by the emulsion electrospinning method. Prolonged yield of EGF upon the material destruction and its biological effect on the MCF7 cell line proliferation have been estimated using ELISA and iCEL-Ligence real-time cell analysis for respectively.
Genes & Cells. 2017;12(4):47-52
pages 47-52 views

Comparative analysis of efficiency of direct and cell-mediated gene therapy of rats with contusion spinal cord injury

Izmailov A.A., Sokolov M.E., Bashirov F.V., Fadeev F.O., Markosyan V.A., Garifulin R.R., Lisyukov A.N., Kuznetsov M.S., Islamov R.R.


Today we have an inadequate set of methods for treating spinal cord injuries. Gene therapy (direct or cell-mediated) is one of the most promising approache for successfully solving this problem. The present study focused on evaluating the therapeutic efficacy of genes encoding vascular endothelial growth factor (VEGF), glial cell-derived neurotrophic factor (GDNF), angiogenin (ANG), and the neuronal cell adhesion molecule (NCAM) in the model of contusion injury in rats. The therapeutic genes in two combinations (VEGF + GDNF + NCAM and VEGF + ANG + NCAM) either were administered intrathecally, with the help of adenoviral vectors, or on cellular carriers - genetically modified mononuclear cells of human umbilical cord blood. On 30 day after a spinal cord injury, the safety of the myelin fibers of the white matter and the kinematics of the left hindlimb joints in experimental animals were analyzed. Both therapeutic combinations of genes have shown a positive effect on the conduction pathways and kinematics of the joints.
Genes & Cells. 2017;12(4):53-59
pages 53-59 views

Analyis of biological compatibility of polylactide nanofibrous matrix vitalized with cardiac fibroblasts in a porcine model

Chepeleva E.V., Balashov V.A., Dokuchaeva A.A., Korobejnikov A.A., Strelnikov A.G., Lependin S.O., Pavlova S.V., Agladze K.I., Sergeevichev D.S., Pokushalov E.A.


Currently the use of synthetic biodegradable polymers based on polyurethane, polycaprolactone, polylactic and polyglycolic acids structures and their co-polymers is one of the most perspective directions of tissue engineering development. Electrospinning was found as an optimal way to produce nanofibers suitable for building several types of biomaterial scaffolds that are used in cell therapy. This technology allows creating a stable biodegradable and highly biocompatible matrix. In this study we investigate the viability of cardiac fibroblasts cultivated on polymeric nanofibrous scaffolds in vitro and in vivo after implantation in the myocardium of an experimental animal. Polymeric nanofibers were produced on an electrospinning unit. Prepared matrixes were vitalized with cell cultures, received from atriums of several mini-pigs. Cell viability was estimated by the use of XTT based colorimetric assay. Two groups of mini-pigs were selected for this research. The first group underwent a procedure of intramyocardial implantation of a matrix, grown with cardiac cell culture. In the second group a clear polymeric matrix was implanted. Seven days after the procedure animals were sacrificed and fragments of myocardium containing implants were harvested. Frozen sections were prepared immediately, then a standard histological analysis and immunofluorescent staining were performed. Current results can be significant for further development of polymeric scaffolds and for research of biophysical and electrophysiological features of cardiac cell cultures, what will help to expand the abilities of contemporary regenerative medicine and may become a standard of autological biological therapy.
Genes & Cells. 2017;12(4):62-68
pages 62-68 views

Bioluminescent monitoring of rat cardiosphere-derived cells in platelet gel engraftment in ischemic heart

Pavlova S.V., Leonova E.A., Chepeleva E.V., Dokuchaeva A.A., Sergeevichev D.S., Pokushalov E.A.


The success of cell therapy depends on an effective method of cell delivery and engraftment. The use of transgenic cells carrying a reporter system based on the luciferase gene allows to perform the quantitative evaluation of the transplantation efficiency in dynamics using biochemical methods. The purpose of this work was to monitor the persistence of rat cardiosphere-derived cells (CDC) after allogeneic transplantation into the periinfarction zone. Transplantation was performed by intramyocardial injection of a cell suspension in a culture medium or in platelet rich plasma (PRP). When injected into the myocardium PRP forms fibrin clots which serves as a matrix for the transplanted cells. The cells were modified by the luciferase enzyme gene by transduction with lentiviruses (CDC-Luc). The activity of luciferase was determined in protein extracts of the myocardium at different time points after the transplantation. It was shown that in the first hour after injections, CDC-Luc is quantitatively detected in the peri-infarction zone irrespective of the use of platelet gel or medium, and their amount does not decrease within 48 hours. During this period, we found a positive effect of the fibrin matrix on the cells - the luminescence of CDC-Luc protein extracts in the platelet gel composition was significantly higher. We suggested that the platelet gel promotes a more favorable microenvironment for CDC-Luc and facilitates the adaptation of cells after transplantation, what reflected in the recovery of the level of luciferase production in cells. Further, we found negative dynamics: CDC-Luc injected in the culture medium is retained in the myocardium for 5 days and on the seventh day their presence is not determined, CDC-Luc in the fibrin matrix is retained in the myocardium for 10 days after transplantation. Thus, despite the successful transplantation of CDC, the integration of cells into the myocardium does not occur. Nevertheless, the use of platelet gel prolongates the time of CDC persistence in the tissue and enhances of their paracrine effect. The use of fibrin matrix can be useful for long-lived cells, such as cardiomyocytes, in particular, to improve the efficiency of transplantation of the tissue engineering biological pacemaker. A luciferase reportering system can be effective for in vitro and in vivo monitoring of cell fate, both in biotechnological stages of cultivation and assembly of the tissue engineering biopee maker, and after myocardial transplantation. In the future, the developed methodological approach will be used to study of tissue-engineering biopacemakers in experimental animals.
Genes & Cells. 2017;12(4):69-75
pages 69-75 views

Proliferative processes and features of tumor cell receptor apparatus of the breast carcinoma

Sazonov S.V., Brilliant A.A., Brilliant Y.M.


Cell division in tumor tissue is associated not only with the growth rate of the tumor, but also its response to ongoing chemotherapy. The isolated subtypes of breast carcinoma differ significantly in the prognosis of the disease, but up to now the level of proliferative processes is only taken into account when dividing the Luminal variants. The aim of the study was to analyze the relationship between the level of proliferative processes in the tumor tissue of breast carcinoma in different tumor subtypes, analyze the patterns of proliferation activity and the characteristics of the receptor apparatus of cells. Material investigated 672 cases of breast invasive carcinomas. In all cases investigated by immunohistochemistry proliferative activity index Ki-67 tumor, as well as steroid hormone receptors ER, PR, and HER-2 / neu receptor. All the cases were divided into biological subtypes according to the standard classification. Mid proliferation index Ki-67 group luminal subtype tumors equaled 8,4±0,2%, in the luminal (HER-2 positive) subtype - 28,8±2,5%, in the luminal (HER-2 negative) subtype - 32,3±1,0%, HER-2 positive subtype - 39,0±2,2%, triple negative subtype - 54,6±2,1% tumors (p <0,05), with the exception of groups in luminal tumors (HER-2 negative) and In the luminal (HER-2 positive) biological subtypes (p> 0,05). Significant differences in proliferation between all treatment groups. The lowest level of proliferation characteristic luminal subtype of breast carcinoma (8.4%), most proliferating tumors are triple negative subtype (Ki-67 = 54.6%). The results show low proliferation of tumors in luminal A subtype of breast carcinoma, as well as a high tumor proliferation in such biological subtypes as luminal B (HER2-positive), luminal B (HER2-negative), HER2 positive and triple negative, and the highest level of expression of Ki-67 observed in carcinomas of triple negative subtype. The observed differences in proliferation levels determined by the expression level of Ki-67 between the subtypes of the breast cancer associated with differences in the receptor apparatus of tumor cells allow us not only to clarify the intracellular mechanisms of cell proliferation in tumor tissue, but also to recommend taking them into account as an additional diagnostic criterion for matching the level Proliferation of the receptor status of tumor cells within each of the subtypes of the breast cancer.
Genes & Cells. 2017;12(4):76-81
pages 76-81 views

Histopathological changes of bone marrow in patients with autosomal recessive osteopetrosis and mutation in TCIRG1 gene

Plaksa I.L., Charlanova E.M., Kravcova V.M., Borovkova A.S., Peshkov M.V., Zubarovskaya L.S., Deev R.V., Afanasiev B.V.


Osteopetrosis is a group of rare hereditary diseases, the general structural manifestation of which is the excessive volume of bone tissue due to violation of osteoclastic resorption. The only way to treat this group of patients so far remains transplantation of hematopoietic stem cells, but the degree of its effectiveness largely depends on the severity of morphological changes in the hematopoietic microenvironment in the bone marrow. In this regard, a comprehensive clinical and morphological analysis, in conjunction with the results of transplantation can help in determining the prognosis of the disease depending on the genetic type of osteopetrosis. The material for the studies were biopsies and smears of bone marrow of patients with osteopetrosis, who were received at the R. Gorbacheva Memorial Research Institute for Pediatric Oncology, Hematology and Transplantation from the republics of Chuvashia and Mari El for carrying out transplantation of hematopoietic stem cells. The histological preparations were stained with hematoxylin and eosin, azur by Romanovsky and by Van Gieson. Bone marrow smears were stained by Romanovsky-Giemsa. The presence of the mutation c.807 + 5G>A in the gene TCIRG1 in patients with autosomal recessive osteopetrosis causes a complete loss of osteoclastic resorption, which is accompanied by pronounced early structural changes in the hematopoietic microenvironment already at the time of diagnosis. This is manifested by an excessive amount of lamellar bone at the same time as the almost complete obliteration of the medullar lacunae, as well as fibrosis of the bone marrow stroma. A common morphological phenomenon among this group of patients is the presence in the bone marrow of an excessive number of osteoclast differentiation cells without the formation of resorption lacunae. This causes a complete suppression of hemopoiesis, which clinically manifests cytopenia and the formation of foci of extramedullary hematopoiesis. Such pronounced structural changes in the hematopoietic microenvironment result in a high risk of primary graft failure during after transplantation of bone marrow hematopoietic stem cells, the effectiveness of which decreases as the child's age increases. Early development of pronounced structural changes in the hematopoietic microenvironment in patients with TCIRG1-mediated osteopetrosis determines the need for diagnosis and transplantation as soon as possible after birth.
Genes & Cells. 2017;12(4):82-90
pages 82-90 views

Medical education at the crossroads. How to step into the future of medicine and medicine of the future?

Kiyasov A.P., Deev R.V., Kiyasova E.V., Gumerova A.A.


Global changes in recent decades, caused by scientific and technological breakthroughs, which are characterized as a genomic revolution and the fourth industrial revolution, raise new pressing issues both for society as a whole and for public health. The latter can not exist and develop without a qualitative, and most importantly, modern medical education, in which significant changes are also taking place. In particular, the transition to standards by the World Federation of Medical Education (WFME) and the Association of Medical Education in Europe (AMEE) - «International Standards of Medical Education for Improving the Quality of Health» (Copenhagen, March 15-19, 2003). In this article, based on the analysis of trends in the development of health care and medical education, possible options for correcting approaches in the teaching of morphological disciplines (normal anatomy, histology, pathology) in medical universities are considered.
Genes & Cells. 2017;12(4):91-96
pages 91-96 views

Legal regulation of biomedical cell products authorization and marketing in international practice

Aleksandrushkina N.A., Tarasova E.V., Makarevich P.I., Gabbasova L.A., Akopyan Z.A., Kamalov A.A., Tkachuk V.A.


Scientific progress and advances in medicine and biology have contributed to emergence of new treatments based on delivery of living cells, genetic material and tissue engineering. The Federal Law N180-FZ “On Biomedical Cell Products” has taken effect in Russia opening a new stage in development of biomedical technologies, the main objective of which is to create a unified harmonious environment for the development, production and sale of safe and effective cell therapy products. At the same time, the system of legal regulation of the market of cellular products abroad has been functioning for more than 10 years and continues to develop actively. This review analyses international experience in regulation of medical biotechnologies market and identifies main features and current problems of existing scheme of biomedical products marketing as well as highlights main issues to be addressed while forming regulatory and legal filed for biomedical cellular products (BMCP) in Russia.
Genes & Cells. 2017;12(4):97-108
pages 97-108 views

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