Department of Human Genetics

Head

Lubov L. Lukash

Professor, Dr. Sci. (Mol. Genetics)
Phone: (380-44) 526-07-39;
Fax: (380-44) 526-07-59;
E-mail: lukash@imbg.org.ua
Web-site:www.lukash.org.ua

Education and Degrees:

1967–1972 Graduate Student, Sumy State Pedagogical University named after A. S. Makarenko, Sumy, Ukraine, M.Sc. (biology and chemistry)

1972–1975 Ph.D. Student, Institute of Molecular Biology and Genetics (IMBG), NASU, Kyiv, Ukraine

1980 Ph.D. (genetics)

1990 Scientific degree of Senior Research Scientist (genetics)

1999 Dr.Sci. (molecular genetics)

2010 Professor (molecular genetics)

Professional Employment:

1972 Laboratory Assistant, Department of Botany, Sumy State Pedagogical University named after A. S. Makarenko, Sumy, Ukraine

1976–1980 Engineer, IMBG NASU, Kyiv, Ukraine

1980 Leading Engineer, IMBG NASU, Kyiv, Ukraine

1980–1982 Junior Research Scientist, IMBG NASU, Kyiv, Ukraine

1982–1986 Senior Research Scientist, IMBG NASU, Kyiv, Ukraine

1986–1990 Leading Research Scientist, IMBG NASU, Kyiv, Ukraine

since 1990 Head of the Department of Human Genetics, IMBG NASU, Kyiv, Ukraine

Honours, Prizes, Awards:

1982 Medal “In memory of the 1500 anniversary of the city of Kyiv”

1998 Diploma of honour of Ministry of Education and Science of Ukraine

2003 Diploma of honour and valuable gift from mayor of Kyiv

2006 Gershenson Award of National Academy of Sciences of Ukraine

2010 Diploma of honour of Vavilov Society of geneticists and breeders of Ukraine

2010 Award of the National Academy of Sciences of Ukraine “For professional achievements”

Research Area:

Research of the biological mutagenesis and role of reparative systems in the correction of genetical damages in cells of pro- and eukaryotic origin

Development of biotechnologies using human stem cells

Ñurrent Research Activities and Recent Achievements:

The influence of exogenous biological factors on mutation process.

In was found that exogenous viruses, DNAs and some mitogenic proteins are able to influence on the spontaneous mutation process and mutagenesis induced by alkylating compounds in mammalian cells in vitro. More detailed investigations were performed to study the possibility of mutation process regulation by impact on DNA repair some mitogen proteins and inhibitors. It was shown jointly with American scientists that a modified base O6-benzylguanine significantly strengthened the mutagenic effect induced by nitrosoguanidine, by reducing enzymatic activity of O6- methylguanine-DNA methyltransferase (MGMT) which plays one of the key roles in cancer cells resistance to alkylating chemotherapeutic drugs (Fig. 1). Currently, together with the Department of Biomedical Chemistry of IMBG NASU, a new generation of MGMT inhibitors are being developed.

Fig. 1. Enhancing of MNNG mutagenic effect in the presence of O6-benzylguanine in cells with different repair systems activity
The regulation of the expression of the gene for repair enzyme MGMT.

We have shown that exogenous cytokines LIF, SCF, IL-3, EMAP II and IFN-a2b (a substrate of preparation “Laferobion”) are able to influence on the MGMT gene expression at the protein level. EMAP-II showed an ability to modulate the level of MGMT gene expression in populations of human cells in vitro differently depending on the experimental conditions. Cytokines LIF, SCF, IL-3 and “Laferobion” usually were causing downregulation of MGMT gene expression in studied human cells. We detected unknown ~50kDa protein which is induced by stress factors as MGMT and recognized by monoclonal anti- MGMT antibodies (clone MT23.3, “Novus Biologicals”, USA). A search for new regulatory elements within the promoter of the mouse and human MGMT gene was performed to learn the complex molecular mechanisms of regulation of this gene expression at the transcriptional level, what perhaps will explain the variation in its expression. Among the detected regulatory promoter sites are those that potentially interact with inducible and tissue-specific transcription factors and alter the level of gene expression in response to various factors. Based on the obtained results we offered the hypothesis about the possiblility of the MGMT gene regulation by various biologically active agents, which are commonly used in supporting therapy of cancer (Fig. 2).

Fig. 2. Regulation of MGMT gene expression on protein level by biologically active compounds in human cells
Investigation of the disruptions of signaling and structural functions of adherent junctions proteins as potential mechanism for some heart pathologies development.

With using conditional knock-out and transgenic animal models we have shown that the structural role of heart adherent junctions is crucial for early cardiogenesis of mammals: loss of N-cadherin in the embryonic heart leaded to the disruption of its development and to embryonic lethality. Cardiospecific deletion of one allele of b-catenin gene resulted in the delay of development of the adult heart, increased expression of the set of embryonic genes (ANP, BNP, b-MHC) and decreased activity of WNT-b-signaling pathway in mutant animals compared with control ones; at the same time morphological reconstructions of adult myocardium were not detected (Fig. 3).

Fig. 3. Cardiac-specific deletion of N-cadherin gene leads to cardiomyocyte adhesion defect and embryonic lethality
The technologies of obtaining, cultivation and differentiation of human stem cells with the purposes of further application in cell therapy.

We processed the approaches to obtaining lines of undifferentiated mammalian cells and directing them into differentiation by using the original method of medium conditioning and specific cytokines and growth factors. A several immortalized cell lines were obtained, which originated from human and mouse stem and progenitor cells: line 4BL derived from peripheral blood of adult donor, line SK1 – from skin of adult patient; lines G1, G4, G6 and G7 – from embryonic primordial mouse gonads. Cell line 4BL was used for creating skin equivalents, which were successfully tested in clinic on a limited cohort of patients with burn disease. Currently, together with the Institute of Cell Therapy we are developing the technologies of obtaining multipotent mesenchymal stromal cells (MMSK) and hematopoietic progenitor cells from native and cryopreserved human placenta (Fig. 4).

Fig. 4. 4BL cells growth features. Cells can growth as monolayer (a) and with forming of compact colonies (b). For production of dermal skin equivalents for thermal burns treatment cells were cultivated on synthetic (c) and natural (d) membranes
Approaches to improvement of biosystems stability to environmental factors and occupational hazards.

We were the first to investigate the effects of some proteins of different origin especially lectins as factors modulating the mutation and repair processes with using pro- and eukaryotic systems. The influence of several lectins of plant and animal origin on the induction of primary DNA damage and gene mutations caused by mutagens with different mechanisms of action (salts of heavy metals and alkylating compounds) was revealed in cell populations and individual mammalian cells in vitro. Gene-protector properties of Sambucus nigra bark lectin and antimutagenic properties of the same lectin and lectin from perch roe were detected and characterized (Fig. 5).

Fig. 5. Modulation of NiCl2-induced DNA damage by black elderberry bark lectin. The comet assay was used
Creation of a model of mutation process and prototype of software for working with databases.

The analysis of experimental data of mutation process manifestation in cell populations treated by different biological factors was performed. The dependence of a mutation process on heterogeneity of populations, energy ensuring, reparative capabilities was shown. In collaboration with the Institute of Cybernetics of V. M. Glushkov NAS of Ukraine the computational modeling of mutagenesis dynamics was carried out, based on obtained facts and hypothesis. As a result, the possibility to predict the effects of mutagenesis in time have occurred, based on empirically derived characteristics of populations; as well as to establish the data bank, reducing time and financial costs on conducting investigations. The software for the calculation of experimental data was also developed. (Fig. 6).

Fig. 6. Co-modeling of mutagenesis processes and repair enzyme expression at consecutive stages of G1 murine cell line in vitro formation. CV – coefficient of variation for chromosome number distribution, index of chromosomal instability level

National Grants:

Projects of National Academy of Sciences of Ukraine:

  • 2012–2016 N 2.2.4.21 Project: “Relation of genotypephenotype in malignant tumors. Modeling of optimal schemes of supporting therapy during the treatment of cancer with alkylating agents” (scientific supervisor – Lukash L. L.)
  • 2010–2014 N 40/2011 Project: “Development of fundamental basis of stem cell therapy of heart pathologies” (scientific supervisor – Lukash L. L.)

International Grants:

  • 2011–2014 7th Framework Programme (FP7) FP7- INCO-2011-6, ERA-WIDE Project: “Strengthening cooperation in Molecular Biomedicine between EU and UKRAINE”, COMBIOM (scientific supervisor – Prof. A. Elskaya; supervisor of a subsection of the project – Lukash L. L.)
  • 2008–2012 RECOOP HST (Regional Cooperation for Health, Science and Technology) Consortium. Lukash L. L. was permanent representative of IMBG NASU in RECOOP HST Consortium.

Collaboration:

with Ukrainian organizations:

  • P. L. Shupyk Natinal Medical Academy of Postgraduate Education (Kyiv)
  • Institute of Cell Therapy (Kyiv)
  • M. M. Amosov National Institute of Cardiovascular Surgery, NAMSU (Kyiv)
  • State Institution “Institute of Neurosurgery named after A. P. Romodanov of NAMS of Ukraine” (Kyiv)
  • National Technical University of Ukraine “Kyiv Polytechnic Institute” (Kyiv)
  • Bogomoletz Institute of Physiology, NASU (Kyiv)
  • Zabolotny Insititue of Microbiology and Virology, NASU (Kyiv)

with foreign organizations:

  • International Institute of Molecular and Cell Biology (Warsaw, Poland)
  • Michigan State University (East Lansing, USA)
  • Pennsylvania State University (University Park, USA)
  • Institute of Toxicology, University Medical Center (Mainz, Germany)
  • Max Planck Institute for Heart and Lung Research (Bad Noyhem, Germany)

Selected publications:

  1. Kochubei T., Kitam V., Maksymchuk O., Piven O., Lukash L. Possible mechanisms of Leukoagglutinin induced apoptosis in human cells in vitro. Cell Biology International.- 2016, Sep. 15
  2. Palchevska O.L., Macewicz L. L., Piven O. O. A link between β-catenin and hypertrophy: evaluation and meta-analysis. Biopolymers and Cell. 2016, 32(2):150-157.
  3. Gryschenko A.A., Bdzhola V.G., Balanda A.O., Briukhovetska N.V., Kotey I.M., Golub A.G., Ruban T.A., Lukash L.L., Yarmoluk S.M. Design, synthesis and biological evaluation of N-phenylthieno [2,3-d]pyrimidin-4-amines as inhibitors of FGFR1. Bioorganic and Medicinal Chemistry. 2015, 23:2287-2293.
  4. Gryschenko AA, Bdzhola VG, Balanda AO, Briukhovetska NV, Kotey IM, Golub AL, Lukash LL, Yarmoluk SM. Design, synthesis and biological evaluation of N-phenylthieno [2,3-d]pyrimidin-4-amines as inhibitors of FGFR1. Bioorganic & Medicinal Chemistry. 2015;23:2287-2293.
  5. Karpova IS, Lylo VV, Macewicz LL, Kotsarenko KV, Ruban TP, Palchykovska LG, Lukash LL. Lectins of Sambucus nigra as biologically active and DNA-protective substances. Acta horticulturae. 2015;1061:93-102.
  6. Lylo VV, Karpova IS, Kotsarenko KV, Macewicz LL, Ruban TP, Lukash LL. Lectins of Sambucus nigra in regulation of cellular DNA-protective mechanisms. Acta horticulturae. 2015;1061:103-108.
  7. Kochubei TÎ, Maksymchuk OV, Lukash LL. Isolectins of phytohemagglutinin are able to induce apoptosis in HEp-2 carcinoma cells in vitro. Experimental oncology. 2015;37(2):116-119.
  8. Lylo VV, Karpova IS, Kotsarenko KV, Macewicz LL, Ruban TA, Dziuba GV, Lukash LL. Isolectins from Sambucus nigra flowers and their effect on MGMT and p53 proteins amount in human cells in vitro. Factors in experimental evolution of organisms. 2015;16:273-276.
  9. Piven OO, Palchevska OL, Lukash LL. Role of Wnt/b-Catenin Signaling in Embryonic Cardiogenesis, Postnatal Formation and Reconstruction of Myocardium. Cytology and genetics. 2014; 48(5):333-343.
  10. Kotsarenko KV, Lylo VV, Macewicz LL, et al. Influence of some biologically active substances on amount of MGMT and MARP proteins in human cells in vitro. Biopolymers and Cell. 2014;30(3):203–208.
  11. Macewicz LL, Lylo VV, Karpova IS, et al. Plant and animal lectines as modulators of MGMT and MARP gene expression in vitro. Factors in experimental evolution of organisms. 2014; 15:260-264.
  12. Podgorsky VS, Kovalenko EA, Karpova IS, et al. Extracellular lectins from saprophytic strains of bacteria of the genus Bacillus (Review). 2014; Applied Biochemistry and Microbiology (Moscow) 50(3):228-234.
  13. Podgorsky V, Kovalenko E, Karpova I, et al. New extracellular Bacillus subtilis lectins with sialic acid specificity. Journal of Agricultural Science and Technology (USA). 2014:541-546.
  14. Babič M, Horák D, Lukash L, et al. Influence of surface-modified maghemite nanoparticles on in vitro survival of human stem cells. Beilstein J. Nanotechnol. 2014;5:1732–1737.
  15. Shablii VA, Kuchma MD, Kyryk VM, et al. Mesenchymal and trophoblast immunophenotype of multipotent stromal cells from human placenta. Biopolym. Cell. 2014; 30(2):118-121.
  16. Kotsarenko KV, Lylo VV, Ruban TP, et al. Influence of IFN-α2b, EMAP II and their medicinal preparations on the MGMT protein amount in human cells in vitro. Biopolym.Cell. 2014; 6.
  17. Kotsarenko KV, Stoliar OA, Lylo VV, et al. DNA repair in MGMT- proficient and MGMT- deficient human cells in vitro. The Ukrainian Biochemical Journal. 2014;86(5):90.
  18. Lukash LL. Regulation of mutagenesis by exogenous biological factors in the eukaryotic cell systems. Biopolym. Cell. 2013; 29(4):283–94. doi:10.7124/bc.000823
  19. Kotsarenko KV, Lylo VV, Macewicz LL, Babenko LA, Kornelyuk AI, Ruban TA, Lukash LL. Change of the MGMT gene expression under influence of exogenous cytokines in human cells in vitro. Cytol. Genet. 2013;47(4):9–15. doi:10.3103/S0095452713040087
  20. Iatsyshyna AP. Current approaches to improve the anticancer chemotherapy with alkylating agents: state of the problem in world and Ukraine. Biopolym. Cell. 2012;28(2):83–92. doi: 10.7124/bc.000032
  21. Shabliy VA, Kuchma MD, Kirik VM, Onishchenko AN, Lukash LL, Lobyntseva GS. Cryopreservation of human placental tissue as a source of hematopoietic progenitor cells and multipotent mesenchymal stromal cells. Cell transplantation and tissue engineering. 2012;7(1):54–62.
  22. Piven OO, Kostetskii IE, Macewicz LL, Kolomiets YM, Radice GL, Lukash LL. Requirement for N­cadherin­catenin complex in heart development. Exp Biol Med (Maywood). 2011;236(7):816–22. doi:10.1258/ebm.2011.010362
  23. Lukash LL. Cell therapy of heart pathologies. Biotechnology. 2008;1(1):40–45.
  24. Pidpala OV, Iatsyshyna AP, Lukash LL. Mobile genetical elements of human genome: distribution and functional role. Cytol. Genet. 2008;42(6):69–81. doi:10.3103/S009545270806011X
  25. Karpova IS, Korets'ka NV, Pal'chykovs'ka LH, Nehruts'ka VV. Lectins from Sambucus nigra L inflorescences: isolation and investigation of biological activity using procaryotic testsystems. Ukr Biokhim Zh. 2007;79(5):145–52.
  26. Macewicz LL, Suchorada OM, Lukash LL. Influence of Sambucus nigra bark lectin on cell DNA under different in vitro conditions. Cell Biol Int. 2005;29(1):29–32. doi:10.1016/j.cellbi.2004.11.007
  27. Lukash LL, Boldt J, Pegg AE, Dolan ME, Maher VM, McCormick JJ. Effect of O6alkylguanineDNA alkyltransferase on the frequency and spectrum of mutations induced by NmethylN'nitroN nitrosoguanidine in the HPRT gene of diploid human fibroblasts. Mutat Res. 1991;250(12):397–409. doi: 10.1016/0027-5107(91)90196-U