Laboratory of Biotechnology

Vladimir A. Riсhter,
Ph.D.
Phone: +7(383)363-51-90, +7(383)363-51-52
Email

• Investigation of human biological fluids proteins and peptides, inducing apoptotic death of cancer cells. Creation of novel therapeutic substances on the basis of recombinant analogues of proapoptotic peptides.
• Investigation of structures and biological functions of extracellular human RNAs.

• Guaranteeing of the Institute activities dealing with radioactive isotopes.
• Labeling of organic compounds with tritium.
• Synthesis of [32P] labeled mono-, di-, triphosphates and oligonucleotides.
• Synthesis of standard and modified nucleosidetriphosphates.

• Human milk protein - lactaptin, inducing the apoptotic death of human adenocarcinoma MCF-7 cells, was isolated and characterized. [Nekipelaya V.V. et al., Dokl. Biochem. Biophys. 2008. 419, 268; Patent RF N.2317304, 2008].
• Engineered producers of lactaptin and lactaptin analogues were obtained and characterized. The specific proapoptotic effect of lactaptin analog RL2 on human cancer cells in culture and on primary cells of human breast tumor was determined. At the same time, non-malignant human mesenchymal stem cells (MSC) showed complete resistance to RL2. [Semenov D.V. et al., The Protein Journal. 2010. 29(3), 174; Patent RF N.2401307, 2010; Fomin A.S. et al., Russian Journal of Bioorganic Chemistry. 2012. 38(1), 1-7].
• The treatment method for mice malignant tumors with lactaptin recombinant analogue RL2 was developed. [Koval O.A. et al., Biochimie. 2012. 94(12), 2467-2474; Patent RF N.2461566, 2012].
• Preclinical studies of antitumor medical drug based on lactaptin recombinant analogue RL2 is now in progress. [State contract No 16.N08.12.1009 (2011-2013)].
• The analysis of blood plasma small noncoding RNA diversity of healthy donors and patients with non small cell lung cancer via massively parallel sequencing technology on SOLID platform was performed. Changes of human extracellular transcriptom supervenient malignant tumor formation were characterized. [Dmitry V. Semenov et al., Expert Opin. Biol. Ther. 2012. 12(1), 43].
• Artificial analogues of U24 snoRNA directed to nucleotides of 28S and 18S rRNA, as well as pre-mRNA and mature mRNA of human heat shock cognate protein hsc70 (gene HSPA8) were designed and synthesized. It was shown that snoRNA analogues penetrate into cells and accumulate in cytoplasm and nucleus. Transfection of cultured human cells with artificial box C/D snoRNA targeted to pre-mRNA induced partial splicing impairments. Transfection with artificial snoRNA directed to 18S and 28S rRNA nucleotides significant for ribosome functioning was found to induce decreasing of MCF-7 cells viability. [Stepanov G.A. et al., Acta Naturaе. 2012. 4(1), 26; Stepanov G.A. et al., BioMed Research International. 2013 (In press)].
• Comparative analysis of blood plasma RNA of healthy donors and patients with non small cell lung cancer allows to characterize changes of human extracellular transcriptom supervenient malignant tumor formation. [Dmitry V. Semenov et al., Expert Opin. Biol. Ther. 2012. 12 (1), 43-51].

1. Koval O.A., Tkachenko A.V., Fomin A.S., Semenov D.V., Nushtaeva A.A., Kuligina E.V., Zavjalov E.L. and Richter V.A. Lactaptin induces p53-independent cell death associated with features of apoptosis and autophagy and delays growth of breast cancer cells in mouse xenografts. PLoS One 2014. 9(4): e93921.
2. Savel’eva A.V., Semenov D.V., Stepanov G.A., Baryakin D.N., Kuligina E.V., Rabinov I.V., Koval O.A., Richter V.A. Effect of recombinant Nucleophosmin 1 Analogue on Artificial RNA Internalization into Human Adenocarcinoma MCF-7 Cells. Russ. J. of Bioorganic Chemistry 2014. 40(1), 48-55.
3. Baryakin D.N., Semenov D.V., Savel’eva A.V., Koval O.A., Rabinov I.V., Kuligina E.V., Richter V.A. Alu and 7SL RNA Analogue Suppress MCF-7 Cell Viability through Modulating the Transcription of Endoplasmic Reticulum Stress Response Genes. Acta Naturae. 2013. 5(4), 92-103.
4. Fomin A.S., Matveev L.E., Matveeva V.A., Richter V.A. Production of monoclonal antibodies to the recombinant analog of lactaptin. Russian Journal of Biopharmaceuticals. 2013. 5(5), 38-43.
5. Stepanov G.A., Semenov D.V., Savelyeva A.V., Koval O.A., Kuligina E.V., Rabinov I.V., and Richter V.A. Artificial box C/D RNAs affect pre-mRNA maturation in human cells. BioMed Research International. 2013; 2013:656158.
6. Koval O.A., Fomin A.S., Kaledin V., Semenov D.V., Potapenko M.O., Kuligina E.V., Richter V.A. A novel pro-apoptic effector lactaptin inhibits tumor growth in mice models. Biochimie. 2012. 94(12), 2467-2474.
7. Fomin A.S., Koval O.A., Semenov D.V., Potapenko M.O., Kuligina E.V., Kit Yu.Ya., and Richter V.A. Analysis of Biochemical Markers of MCF-7 Cell Apoptosis Induced by a Recombinant Analogue of Lactaptin. Russian Journal of Bioorganic Chemistry. 2012. 38(1), 77–82.
8. Semenov D.V., Baryakin D.N., Brenner E.V., Kurilschikov A.M., Vasiliev G.V., Bryzgalov L.A., Chikova E.D., Richter V.A. Unbiased approach to profile the variety of small non-coding RNA of human blood plasma with massively parallel sequencing technology. Expert Opin. Biol. Ther. 2012. 12 (1), 43-51.
9. Stepanov G.A., Semenov D.V., Kuligina E.V., Koval O.A., Rabinov I.V., Kit Y.Y., Richter V.A. Artificial Human Box C/D Small Nucleolar RNA Analogues as Regulators of Alternative Splicing of pre-mRNA-target. Acta Naturaе. 2012. 4(1), 26-35.
10. Gruntenko N.Е., Bogomolova E.V., Adonyeva N.V., Karpova E.K., Menshanov P.N., Alekseev A.A., Romanova I.V., Li S., Rauschenbach I.Yu. Decrease in juvenile hormone level as a result of genetic ablation of the corpus allatum cells affects the synthesis and metabolism of stress related hormones in Drosophila. Journal of Insect Physiology. 2012. 58(1), 49-55.
11. Van der Vaart, Dmitry V. Semenov, Elena V. Kuligina, Vladimir A. Richter, Pieter J. Pretorius. Parallel tagged 454 sequensing for the characterization of circulating DNA. Advances in Experimental Medicine and Biology. 2011, 29-34.
12. Van der Vaart M., Semenov D.V., Kuligina E.V., Richter V.A., Pretorius P.J. Advanced Analysis of Human Plasma DNA Sequences Produced by Parallel Tagged Sequencing on the 454 Platform. P.B. Gahan (ed.), Circulating Nucleic Acids in Plasma and Serum, Springer Science+Business Media B.V. 2011, 35–40.
13. Semenov D.V., Stepanov G.A., Baryakin D.N., Koval O.A., Kuligina E.V., Richter V.A. Extracellular RNA as Regulators of Cellular Processes. P.B. Gahan (ed.), Circulating Nucleic Acids in Plasma and Serum, Springer Science+Business Media B.V. 2011, 233-237.

1. Koval' O.A., Kaledin V.I., Kuligina E.V., Semenov D.V., Fomin A.S., Potapenko M.O., Richter V.A. The method of mammal's tumors treatment. Patent RF N.2461566, 2012.
2. Tikunova N.V., Semenov D.V., Babkina I.N., Kuligina E.V., Koval O.A., Fomin A.S., Matveeva V.A., Matveev L.E., Matveev A.L., Richter V.A. Recombinant plasmid DNA pFK2, providing synthesis of the recombinant peptide which is the analog of human kappa-casein, and recombinant peptide – the analog of human kappa-casein fragment, with the apoptotic activity against human tumor cells. Patent RF N.2401307, 2010.
3. Matveev L.E., Matveev A.L., Semenov D.V., Fomin A.S., Kuligina E.V., Matveeva V.A., Tikunova N.V., Babkina I.N., Richter V.A. Strain of hybridous cultured mammalian cells Mus. Musculus producing monoclonal antibodies specific for the peptide possessed apoptotic activity against human tumor cells. Patent RF N.2402605, 2010.

• N.13-04-01058-a «Structures, functions and diagnostic significance of extracellular RNA within microvesicles, exosomes and ribonucleoprotein complexes of human blood plasma»(2013- 2015)
  
• N.13-04-01313-a «Antitumor recombinant peptide lactaptin: mechanism of cell penetration and apoptosis induction» (2013- 2015)
• N.13-04-01457-a «Immune system activation in antitumor therapy by recombinant analog of lactaptin» (2013- 2015)
• N.14-04-31468 «Molecular mechanisms of artificial small nucleolar RNAs action on human cells» (2014-2015)

• N.84 «Natural microvesicles and complexes transporting nucleic acids: properties, diological functions and prospects applications in diagnostics and therapeutics»

• N.94 «Development of cell technologies for carcinogenesis mechanism study and anti-cancer drugs testing»

N.5 «Basic Science for Medicine»

• N.13 “Development of therapeutic composition of lactaptin with protein preparations for oncological neoplasms combined therapy” (2013-2015)

• Multicolor Real-Time PCR Detection System Bio-Rad, IQ5;
• Real-Time PCR Detection System Roche LightCycler 480 II;
• Amplifier Eppendorf Mastercycler gradient;
• Amplifier Eppendorf Mastercycler;
• French Press, GlenMills;
• Chromatography System Bio-Rad BioLogic LP, Chromatography System GE Healthcare Avant 150;
• Chromatography System Bio-Rad BioLogic Dou Flow;
• Centrifuge Hitachi CR 22 GIII;
• Centrifuge Eppendorf 5810 r, Microscope Axiovert 40 С, Cytometer BD FACS Canto II TM.