Department of Cell Regulatory Mechanisms

Head

Vitalii A. Kordium

Professor, Dr. Sci. (Microbiol.),
Corresponding Member of NASU and
Full Member of Acad.Med.Sci. of Ukraine
Phone: (380-44) 526-55-96;
Fax: (380-44) 526-07-59;
E-mail: kordium@imbg.org.ua

Laboratory of Modification of Biologically Active Compounds

Head: Anatoliy I. Potopalsky,
Ph.D. (Med. Sci.), Senior Lecturer

Laboratory of Microbial Ecology

Head: Natalia O. Kozyrovska,
Ph.D. (Mol. Biol.), Senior Staff Scientist

Education and Degrees:

1950–1955 Graduate Student, Faculty of Biology, Taras Shevchenko National University of Kyiv, Kyiv, Ukraine

1955–1958 ostgraduate Student, Department of Microbiology, Taras Shevchenko National University of Kyiv, Kyiv, Ukraine

1960 Ph.D. (microbiology)

1972 Dr.Sci. (microbiology)

1991 Corresponding Member of NASU (medical genetics)

1993 Professor

2000 Full Member of NAMSU (medical genetics)

Honours, Prizes, Awards:

1977 Prize of Council of Ministers of the USSR

1979 State Prize of Ukraine in Science and Technology for a series of works on space biology

2003 Diploma of Verkhovna Rada of Ukraine

2008 Honoured Worker of Science and Technology of Ukraine

2012 The Order of Prince Yaroslav the Wise, V degree

Research Area:

The study on intercellular informational interactions

Ñurrent Research Activities and Recent Achievements:

The development of gene therapy for diabetes type-1 and atherosclerosis.

A series of recombinant vector DNAs containing the human full-size preproinsulin and apolipoprotein A1 genes in the integrative expression cassettes have been constructed in the Department. The mammalian cell lines that express the target human proteins were obtained. Vector DNA polyplexes based on modified polyethyleneimine containing glycoside ligands have been optimized for efficient and safe targeted gene transfer in mammalian liver cells. The consistently reproducible technique for obtaining experimental models of chemically induced diabetes in animals of four species (mice, rats, rabbits and pigs) has been perfected. The models of alimentary hypercholesterolemia and of lipid metabolism disorders under diabetes, induced by streptozotocin in rabbits, have been developed. Therapeutic efficacy of targeted vector DNAs has been demonstrated in long-term experiments with these animal models (Fig. 1): a) antihyperglycemic action of polyplexes containing human preproinsulin gene in the model of type 1 diabetes in mice, rats, rabbits and pigs; b) antiatherogenic effects of polyplexes containing human apolipoprotein A1 gene in the models of alimentary hypercholesterolemia and combined carbohydrate and lipid metabolism disorders in rabbits.

Fig. 1. a – Gene Therapy Correction of Streptozotocin-induced Diabetic Hyperglycemia in Landras Swine, b – Gene Therapy for Experimental Hypercholesterolemia in Rabbits
Study on human umbilical cord MSC isolation, cultivation, characterisation and usage.

A technology of MSC isolation from human umbilical cord Wharton jelly with maximal preservation of their native characteristics has been developed. The necessity of individual approach for the cells obtaining from umbilical cord was confirmed. The optimal culturing conditions, in particular, considering an influence of gas mixture were determined. It has been shown that lowered oxygen concentration and nitrogen substitution with argon had positive effect on the umbilical cord MSC during their cultivation. The morphological characteristics as well as surface markers expression were studied. MSC from umbilical cord were shown to be heterogeneous at isolation (0 passage), but already at the first passage they had typical for MSC spindle-shaped morphology. They were CD73, CD90, CD105 positive and CD34, CD45 negative, which is characteristic of MSC. After the second ex vivo passage (Fig. 2), gradual change of the MSC properties occurred (cell morphology, surface markers expression, doubling time, etc.), as well as spontaneous differentiation to adipo- and chondrocytes began. Accordingly, only the cells of this or lower culture level passage can be used. Collagens I and II types were extracted, and matrices for the cell cultivation were prepared.

Fig. 2. MSC culture, 2 passage. Confocal microscopy, staining with ethidium bromide + FITC

In recent years, the use of transplanted cells producing active factors has been proven to be an emergent technology. However, the transplantation of allogenic and xenogenic cell material results in the immune rejection of cell transplants. Cell microencapsulation is a promising tool to prevent an attack of immune system even in case of xenogenic transplants. Microcapsules, prepared from semipermeable membranes, ensure transmission of target proteins, on the one hand, and defend the cells from attacks of immune system – on the other. It makes possible to avoid the use of long-term therapies of modulating and/or immunosuppressive agents, which have potentially severe side-effects. Usage of encapsulated cells, producing growth factors, cytokines, hormones and other therapeutic proteins, is at present a very promising way of delivery of therapeutic material into an organism. The Department takes part in this field of research. The genetically engineered cells are a source of therapeutic proteins. Transgenic mammalian cell lines producing some important recombinant human cytokines (LIF, FGF2, IL10) have been obtained by us via a nonviral gene transfer technique. The recombinant protein secretion into the cultural medium was shown. When the cells producing cytokines were encapsulated in alginate microcapsules, the production of the recombinant cytokines was continuing and the cytokine molecules are secreted from the microcapsules into the cultural medium (Fig. 3). The therapeutic effects of the cytokines, produced by genetically modified eukaryotic cells, are planned to be tested on animal models when the microcapsules will be transplanted to animals suffering from some disordes caused by the lack of tested cytokines.

Fig. 3. ÑÍÎ-Ê1 cells transfected by pC1eGFP in alginate microcapsules (à – light microscopy; b – fluorescent microscopy)
The study on influence of recombinant cytokines on structural and functional state of ischemic tissues.

It has been demonstrated in the experimental model of chronic kidney ischemia in rabbits that the intraparenchymal administration of the FGF-2 in developed polymeric carrier, based on cross-linked heparin, 5 months after ligature application, decreases the initial sclerotic changes in stroma and vessels and protects from the development of irreversible sclerotic changes in the interstitium and atrophy of the elements of parenchyma in ischemic and contralateral kidney. The data of angiography indicate that the architectonics of arterial bed of kidney of experimental animals with segmental ischemia was almost the same as of intact contralateral kidney after the injection of the preparation of FGF-2 studied (Fig. 4). The gene coding for human interleukin-10 has been cloned, the high-level expression of this cytokine in E.coli cells has been obtained, and the method for its downstream purification and obtaining in biologically active form has been elaborated in the Department. It has been shown that the intraparenchymal administration of the interleukin-10 into ischemic kidney tissue reduced the activity of the processes of lipid peroxidation and decreased the risk of ischemic kidney damage. The genes coding for human stromal-derived factor-1α (SDF-1α) and vascular endothelial growth factor have been cloned for further studies. The expression of these recombinant proteins in E.coli cells has been obtained, and the method for downstream purification and obtaining in biologically active form of SDF-1α has been elaborated.

Fig. 4. Angiogram of vessels of rabbit urinary system after 6 months the application of ligatures on the upper pole of left kidney. The place of the ligature application shown by arrow

National Grants:

Projects of National Academy of Sciences of Ukraine:

  • 2010–2014 N 35/13 Progect: “Fundamental foundations of multigene therapy for mass pathology with complications” (scientific supervisor – Kordium V. A.)
  • 2010–2014 N 36/10 Progect: “Developing a fundumental basis for obtaining new generation immunoreagents” (scientific supervisor – Okunev Î. V.)

Projects of State Agency on Science, Innovations and Informatization of Ukraine:

  • 2013–2014 Progect: “The Development of technology for the repairing injured liver function by MSC transplantation” (scientific supervisor – Rymar S. Y.)

Collaboration:

with Ukrainian organizations:

  • State Institute of Genetic and Regenerative Medicine, NAMSU (Kyiv)
  • D. F. Chebotarev State Institute of Gerontology, NAMSU (Kyiv)
  • State Institution “V. K. Gusak Institute of Urgent and Reconstructive Surgery of NAMS of Ukraine” (Donetsk)
  • State Institution “Institute of Urology of NAMS of Ukraine” (Kyiv)
  • State Institution “V. P. Komisarenko Institute of Endocrinology and Metabolism of NAMS of Ukraine” (Kyiv)
  • Educational and Scientific Centre “Institute of Biology” Taras Shevchenko National University of Kyiv (Êyiv)
  • State Institution “Institute of Pediatrics, Obstetrics and Gynecology of NAMS of Ukraine” (Kyiv)
  • State Institution “Institute of Neurosurgery named after A. P. Romodanov of NAMS of Ukraine” (Êyiv)
  • State Institution “Institute of Otolaryngology named after O. S. Kolomiychenko of NAMS of Ukraine” (Kyiv)
  • State Institution “National Research Center for Radiation Medicine of NAMS of Ukraine” (Kyiv)
  • State Institution “V. P. Filatov Institute of Eye Diseases and Tissue Therapy of NAMS of Ukraine” (Odessa)
  • Institute of Veterinary Medicine, NAASU (Êyiv)
  • O. V. Palladin Institute of Biochemistry, NASU (Êyiv)

with foreign organizations:

  • Leipzig University (Leipzig, Germany)
  • Centre for Stem Cells Sciences (Hyderabad, India)

Selected publications:

  1. Moshynets O.V., Spiers A.J. Book: "Microbial Biofilms - Importance and Applications", "Viewing biofilms within the larger context of bacterial aggregations", Edited by Dharumadurai Dha-nasekaran and Nooruddin Tha-juddin, ISBN 978-953-51-2436-8, Print ISBN 978-953-51-2435-1, 522 p., July, 2016; Chapter 1:3-22
  2. Pokholenko I., Dubey I., Vozianov S., Romanenko A., Pirogov V., Bazalytska S., Nikitaiev S., Kordium V. Development of the affinity-based delivery system for heparin-binding proteins. The International Journal of Ar-tificial Organs. 2016; 39(7): 373-374
  3. Kordyum VA Clearing flows in the biosphere. And not only. K: Akademperyodyka 2016; 200 p.
  4. Starosyla SA, Volinetc GP, Gorbatiuk OB, Lucashov SS, Golub AG, Bdzhola VG, Yarmoluk SM. Identification of Apoptosis signal-regulating kinase 1 (ASK1) inhibi-tors among the derivatives of Ben-zothiazol-2-yl-3-hydroxy-5-phenyl-1,5-dihydro-pyrrol-2-one. Bioorganic and Medicinal Chemistry. 2015;23:2489–2497.
  5. Reva O, Ovcharenko LP, Kukharenko O, Shpylova S, Podolich O, de Vera J-P, Kozyrovska N. Metabarcoding of the kombucha microbial community grown in different microenviromental. AMB Express. 2015;5(1):35, doi: 10.1186/s13568-015-0124-5
  6. Maslova O, Fedevych O, Shuvalova N, Deryabina O, Zhovnir V, Novak M, Kruzliak P. Morphological characteristics of cultured fresh and thawed pericardium cells. Cell and Tissue banking. 2015; September on-line:1-5, doi: 10.1007/s10561-015-9532-7
  7. Shuvalova NS, Kordium VA. Comparison of proliferative activ-ity of Wharton jelly mesenchymal stem cells in cultures under various gas conditions. Biopolym. Cell. 2015; 31(3):233-239.
  8. Pokholenko IO, Gorbatiuk O, Okunev O, Irodov D, Degtiarova M, Palivoda O, Kordium V. Development of the chroma-tographic medium for the affinity isolation of the recombinant hIFN-β1b based on immobilized singlechain antibodies. Biopolym. Cell. 2015;31(4):279–284.
  9. Gorbatiuk OB, Bakhmachuk AO, Dubey LV, Usenko MO, Irodov DM, Okunev OV, Kostenko OM, Rachkov AE, Kordium VA.Recombinant Staphylococcal protein A with cysteine residue for preparation of affinity chromatography stationary phase and immunosensor applications. Biopolym. Cell. 2015; 31(2):115–122.
  10. Kovalchuk MV, Shuvalova NS, Pokholenko IO, Gulko TP, Deryabina OG, Kordium VA. Monitoring of transplanted human Mesenchymal Stem Cells from Wharton’s Jelly in xenogeneic systems in vivo. Biopolym. Cell. 2015;31(3):193–199.
  11. Rachkov AE, Bakhmachuk AO, Matsishin MJ, Gorbatiuk OB, Khristosenko RV, Ushenin IuV, Soldatkin AP. SPR investigations of the formation of intermediate layer of the immunosensor bioselective element based on the recombinant Staphylococcal protein A. Biopolym. Cell. 2015; 31(4):301–308.
  12. Zaets I, Podolich J, Kukharenko O., et al. Bacterial cellulose may provide the microbial-life biosignature in the rock records. Advances in Space Research. 2014;53(5):828–835.
  13. Boretska M., Datsenko I., Suslova O, et al. Amino acid composition of tightly bound exopolymeric substances produced by corrosion-related bacteria in presence of mild steel. Advances in Microbiology. 2014;4:808–815.
  14. Lukash LL, Ruban TP, Kolomiets YaN, et al. In vitro detection of cytoto-xicity of surface-modified maghemite nanoparticles in human cells. Beilstain J. Nanotechnol. 2014;5:1732–1737.
  15. Sviatenko OV, Gorbatiuk ÎB, Vasylchenko OA. Application of immunoglobulin–binding proteins A, G, L in the affinity chromatography. Biotechnologia Acta. 2014;7(2):34–45.
  16. Slyvka AV, Okunev OV. Molecular mechanisms of versatile biological activity of interleukin-7. Biopolym. Cell. 2014;30(5):349–358.
  17. Pokholenko IaO, Chetyrkina MD, Dubey LV, et al. Development and characterization of porous functionalized collagen scaffolds for delivery of FGF-2. Biopolym. Cell. 2014;30(3):216–222.
  18. Kordium V. A. 40 years within two epochs of two millennia. Biopolym. Cell. 2013; 29(4):255-260 doi:10.7124/bc.00081F
  19. Maslova O, Kordium V, Deryabina O Umbilical Cord matrix cells – promising instrument for regenerative medicine. In: Loredana De Bartolo, Augustinus Bader, editors. Biomaterials for stem cell therapy: state of the art and vision for the future: CRC Press, Taylors&Francis Group, Boca Raton, London, New York, A science publishers book; 2013. p. 212–227.
  20. Maslova OO, Shuvalova NS, Sukhorada OM, et al. Heterogeneity of umbilical cords as a source for MSC. Dataset Papers in Biology 2013, Article ID 370103.
  21. Shpylova SP, Shuvalova NS, Deryabina OG, Kordium VA. Sponta neous formation of spheroids in human umbilical cord matrix cell cultures. Studia Biologica. 2012; 6(2):79–86.
  22. Maslova OO, Shuvalova NS, Sukhorada OM, et al. Alteration of morphofunctional characteristics of umbilical cord matrix me­senchymal cells during passaging. Problems of cryobiology.2012; 22(2):153–6.
  23. Pokholenko IaO, Rachkova LT, Gulko TP, et al. The development of biodegradable scaffold from collagen type I and study of their biological properties in vitro and in vivo. In: Achievements and problems of genetics, breeding and biotechnology. K.: Logos. 2012. p. 358–62.
  24. Rymar SYu, Ruban TA, Irodov DM, Kordium VA. Expression and secretion of human recombinant LIF by genetically modified mammalian cells. Biopolym. Cell. 2011; 27(1):53–8. doi: 10.7124/bc.000082
  25. Vozianov OF, Romanenko AM, Pirogov VO, et al. Novel systems for the restoration of blood flow in ischemic kidney. Ninth International Symposium on Frontiers in biomedical polymers FBPS. 2011:103–4.
  26. Toporova OK, Gulko TP, Sukhorada OM, et al. Longterm correction of experimental diabetes in mice and pigs after nonviral insulin gene therapy. Human Gene Therapy. 2009; 20(4):397.
  27. Novikova SN, Toporova OK, Sukhorada OM, et al. Experimental gene therapy for atherosclerosis. Problemu stareniya i dolgoletiya. 2008; 17(2):174–83.
  28. Toporova OK, Kyrylenko SD, Irodov DM, Kordium VA. Plasmid vector for human preproinsulin gene delivery in mammalin cells. Biopolym. Cell. 2007; 23(2):100–7. doi:10.7124/bc.00075B