научная статья по теме ТЕЗИСЫ МЕЖДУНАРОДНОЙ КОНФЕРЕНЦИИ “MOLECULAR MECHANISMS OF GROWTH AND PROGRESSION OF MALIGNANT NEOPLASMS”, ПОСВЯЩЕННОЙ 55-ЛЕТИЮ ИНСТИТУТА МОЛЕКУЛЯРНОЙ БИОЛОГИИ РАН И 120-ЛЕТИЮ СО ДНЯ РОЖДЕНИЯ В.А. ЭНГЕЛЬГАРДТА (МОСКВА, 11–12 ДЕКАБРЯ 2014 Г.) Биология

Текст научной статьи на тему «ТЕЗИСЫ МЕЖДУНАРОДНОЙ КОНФЕРЕНЦИИ “MOLECULAR MECHANISMS OF GROWTH AND PROGRESSION OF MALIGNANT NEOPLASMS”, ПОСВЯЩЕННОЙ 55-ЛЕТИЮ ИНСТИТУТА МОЛЕКУЛЯРНОЙ БИОЛОГИИ РАН И 120-ЛЕТИЮ СО ДНЯ РОЖДЕНИЯ В.А. ЭНГЕЛЬГАРДТА (МОСКВА, 11–12 ДЕКАБРЯ 2014 Г.)»

УДК 577.2

ТЕЗИСЫ МЕЖДУНАРОДНОЙ КОНФЕРЕНЦИИ "MOLECULAR MECHANISMS OF GROWTH AND PROGRESSION OF MALIGNANT NEOPLASMS", ПОСВЯЩЕННОЙ 55-ЛЕТИЮ ИНСТИТУТА МОЛЕКУЛЯРНОЙ БИОЛОГИИ РАН И 120-ЛЕТИЮ СО ДНЯ РОЖДЕНИЯ В.А. ЭНГЕЛЬГАРДТА

(Москва, 11—12 декабря 2014 г.)

International Conference "Molecular mechanisms of growth and progression of malignant neoplasms" dedicated to the 55th anniversary of the Engelhardt Institute of MolecUlar Biology of the Russian Academy of Sciences and the 120th anniversary of W.A. Engelhardt.

DOI: 10.7868/S0026898415050171

Runxl MUTATIONS: SETTING THE STAGE FOR LEUKEMIC TRANSFORMATION. K. Behrens, B. Niebuhr, U. Müller, M. Ziegler, C. Stocking (Heinrich-Pette-Institute, Leibniz Institute of Experimental Virology, Hamburg, Germany). The Runxl transcription factor is among the most frequently mutated genes in acute myeloid leukemia (AML). In addition to the generation of the RUNX1-RUNX1T1 fusion protein in patients with the translocation (8;21), circa 15% of patients with a normal karyotype harbor missense mutations or frame-shift mutations in the RUNX1 gene, resulting in either a highly compromised protein lacking DNA-bind-ing activity or a null allele. Recent studies have demonstrated that AML transformation normally occurs at the level of the granulocyte-monocyte-progenitor (GMP), thus we sought to dissect the role of Runx1 at this critical stage of myeloid differentiation by examining the hematopoietic progenitor compartment in conditional Runx1 knockout (KO) mice. Earlier studies have demonstrated increased cell numbers in the stem cell compartment in these mice, but a rigorous assessment of myeloid progenitors has not been performed. Our analysis showed that loss of Runx1 resulted in increased levels of all myeloid progenitors, with a 2.2-fold increase in the absolute GMP numbers. Furthermore, Runx1-deficient GMPs gave rise to 40% more colonies than controls, demonstrating increased self-renewal activity within this population. To identify Runx1 target genes that impact at the level of the GMP, expression analysis of Runx1 wild-type and KO GMPs was performed. In addition, the transcriptome of primary KO GMPs genetically engineered to conditionally re-express RUNX1 in vivo was determined before or after Runx1 induction. Genome wide DNA-binding analysis established Runx1 binding to regulatory regions of target genes. This analysis confirmed that Runx1 regulates several stem-cell specific transcription factors and adhesion molecules that are critical for maintaining self-renewal and blocking GM differentiation. In summary, we conclude that Runx1 has at least two critical functions in normal myeloid development. In the early stem cell and progenitor compartment, Runx1 represses genes that mediate adhesion to the stem cell niche, thereby facilitating the differentiation program at the expense of self-renewal. On the other hand, during myeloid maturation, Runx1 augments both G and M developmental programs, presumably

by facilitating up-regulation of critical granulocytic and monocytic genes. We predict that this former function is of critical importance during leukemogenesis, in which reduced levels of functional Runxl increases the number of myeloid progenitors with self-renewal potential, which are critical targets of secondary genetic mutations during evolution of the leukemic clone.

ADENOVIRUS MEDIATED TRANSFORMATION OF PRIMARY HUMAN CELLS: A MODEL SYSTEM TO STUDY VIRUS INDUCED CELL TRANSFORMATION.

T. Speiseder1, D. Indenbriken1, A. Schellenberg2, N. Akyuz2, A. Grundhoff1, B. Fehse3, C. Lange3, T. Dobner1 (1Hein-rich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany; 2Department of Oncology, Hematology, BMT with Section Pneumology, Hubertus Wald Tumorzentrum — University Cancer Center Hamburg, University Hamburg Medical Center Hamburg-Eppendorf, Hamburg, Germany; 3Research Unit Cell and Gene Therapie, University Hamburg Medical Center Hamburg-Eppendorf, Hamburg, Germany). The human adenovirus (Ad) E1A/E1B (E1) and E4 proteins are multifunctional regulators of Ad replication participating in many processes required for maximal virus production and Ad-mediated oncogenesis. Their growth promoting activities correlate with their ability to interact with and to manipulate a number of cellular key regulators controlling transcription, apoptosis, DNA repair and protein stability. Prominent examples are p53, Mre11, DNA ligase IV, factors of the ubiquitin- and SUMO-conjugation machineries as well as components of PML nuclear bodies. Over the past years work in our group has concentrated on the biochemical and genetic analysis of the E1 and E4 proteins in productive Ad-infection and cell transformation, focusing on the mechanisms by which these viral factors promote oncogenic transformation of primary mammalian cells in culture. It has been shown, that tumor induction and focus formation in the rodent system is induced by the interactions of E1A/B and E4 with several host cell factors. However, it is widely believed, that human adenoviruses or at least their E1 oncogenes are not able to transform primary human cells. Even though transformations studies in the rodent system identified a lot of host cell factors involved in the transformation process, there are some differences be-

tween the rodent and the human cell system and data can only be partial transferred from one system to the other. As there are some publications linking human adenoviruses to tumor formation in humans, we developed a model system, based on primary human mesenchymal stromal cells (hMSCs), in order to study the growth promoting activities of adenoviral oncogenes in the human system. Using a lentivirus-based transfer-system to model the classical concept ofvirus-medi-ated transformation, we could show for the first time that primary hMSCs are highly susceptible for transformation by Ad oncogenes. Ad5 E1 transformed hMSCs exhibit properties commonly associated with a high grade of oncogenic transformation. To unravel the underlying molecular mechanisms, which induce the process of cell transformation, we use Illumina-based next generation sequencing (NGS) for our comparative transcriptome analysis to detect novel, yet unknown host cell factors, which are highly deregulated after transduction of Ad E1 oncogenes into these human progenitor cells. Moreover, our data suggest that these pheno-types are, at least in part, due to alterations of the host cell epigenome as well as chromosomal reorganizations of the genome. In fact, there is a good reason to believe that these changes could be a general mechanism contributing to the etiology of many infection-associated cancers, which may also form the molecular basis of other transformation processes.

UNUSUAL FEATURES OF TELOMERASE AND NOVEL TELOMERASE INHIBITORS. T.S. Zatsepin, M.I. Zvereva, M.P. Rubtsova, O.A. Dontsova (Department of Chemistry and Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, 119991 Russia). Telom-erase is a multicomponent complex responsible for the maintenance of telomere length, which consists of special RNA molecules, telomerase reverse transcriptase subunit, and of several other proteins. Telomerase activity is characteristic for cells that have unlimited division potential: germ, stem, embryo, as well as for yeast. Activation of telomerase correlates with the process of carcinogenesis since tumor cells possess unlimited replicative potential. In about 90% of tumor cell types active telomerase is a key participant of telomere length maintenance. Therefore, telom-erase is an attractive target for anticancer therapy. Reduced telomerase activity leads to induction of cellular senescence, and eliminates tumor cells from further active proliferation by apoptosis [1]. Unusual properties of thetelom-erase RNA may interfere with the development of effective anticancer drugs, it is therefore necessary to keep the components of telomerase intact, but to disrupt its activity [2]. We proposed a new approach to telomerase inhibition using it's disruption at the stage of assembly. This approach is based on the usage of bifunctional chimeric oligonucle-otides, which contain two oligonucleotide fragments complementary to the functional domains of telomerase RNA that are connected by non-nucleotides linker in different orientations ( 5' ^ 3', 5' ^ 5' or 3' ^ 3'). Such chimeras (in nm concentrations) inhibit the activity of telomerase in vitro and 100 times more effective in vivo. Such chimeras do not affect the level of telomerase RNA in the cell, non-toxic and inhibit the assembly of active telomerase complex [3]. This work was supported by grants from the Russian Foundation for Basic Research (14-04-01092, 14-04-01637) and Competition for Projects of Fundamental Research (RFBR-OFI, 11-026-12051).

LITERATURE

1. Zvereva M.I., Shcherbakova D.M., Dontsova O.A. 2010. Telomerase: structure, functions, and activity regulation. Biochemistry (Mosc.). 75 (13), 1563-1583.

2. Rubtsova M.P., Vasilkova D.P., Malyavko A.N., Narai-kina Y.V., Zvereva M.I., Dontsova O.A. 2012. Telomere lengthening and other functions of telomerase. Acta Naturae. 4 (2), 44-61.

3. Azhibek D., Zvereva M., Zatsepin T., Rubtsova M., Dontsova O. 2014. Chimeric bifunctional oligonucleotides as a novel tool to invade telomerase assembly. Nucl. Acids Res. 42 (15), 9531-9542.

GENETIC BARCODING TO ASSESS CLONALITY OF MALIGNANT DISEASES. Kerstin Cornils1, Lars Thielecke2, Doreen Winkel-mann1, Tim Aranyossy1, Andreas Dahl3, Svenja Hüser1, Nadja Kleist1, Ingo Roeder2, Ingmar Glauche2, Boris Fehse1 (1Research Dept. Cell and Gene Therapy, Clinic for Stem Cell Transplantation, University Medical Centre Hamburg-Eppendorf, Martinistr. 52, 20246 Hamburg, Germany; 2Institute for Medical Informatics and Biometry, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, 01307 Dresden, Germany; 3Deep Sequencing Group SFB 655, Biotechnology Centr

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