Vorobyev Pavel
Verified @gmail.com
Scopus Publications
- Humanized Murine Glioblastoma Models for Evaluation of Coxsackievirus Oncolytic Therapy
Yana D. Gumennaya, Marat P. Valikhov, Elizaveta R. Naberezhnaya, Pavel O. Vorobyev, Veronika V. Vadekhina, Olga N. Alekseeva, Anastasiia O. Sosnovtseva, Dmitry V. Kochetkov, Alesya V. Soboleva, Leen Ibrahim, Stepan A. Ionov, Gaukhar M. Yusubalieva, Alexander V. Ivanov, Peter M. Chumakov, Anastasia V. Poteryakhina
Cancers, 2026
Background/Objectives: Glioblastoma remains the most lethal primary brain tumor in adults, and progress in oncolytic virotherapy is limited by the lack of immunocompetent models permissive to human-tropic viruses. Methods: Here, murine CT-2A and GL261 glioma and B16 melanoma cell lines were engineered to express human Coxsackievirus and Adenovirus Receptor (CXADR) fused to tagBFP, generating “humanized” tumors that preserve parental growth characteristics while acquiring high susceptibility to group B Coxsackieviruses (CVBs) and adenovirus serotype 5. Results: CXADR expression in CT-2A, GL261, and B16 cells markedly enhanced binding, internalization, and replication of CVBs in vitro, with the strongest effect observed for LEV14 (attenuated CVB5), which reached up to 105-fold higher viral titers in humanized cells compared with parental cells. Unchanged sensitivity to vesicular stomatitis virus indicated receptor-specific effects. Humanized CT-2A-CXADR-BFP and GL261-CXADR-BFP cells initiated aggressive subcutaneous and intracranial tumors in syngeneic C57BL/6 mice without signs of immune rejection, and histology and MRI confirmed invasive high-grade glioma phenotypes. In intracranial CT-2A-CXADR-BFP tumors, repeated intratumoral LEV14 administration induced extensive tumor necrosis and prolonged survival despite the rapid development of neutralizing antibodies. Systemic intravenous LEV14 dosing produced strong oncolytic activity against subcutaneous CT-2A-CXADR-BFP tumors, as demonstrated by pronounced tumor growth inhibition, long-lasting regression in a subset of animals with gliomas, and improved overall survival. Conclusions: Collectively, these data establish CXADR-humanized models as versatile, immunocompetent platforms for evaluation of CXADR-dependent oncolytic enteroviruses. - Establishment of a Panel of Human Cell Lines to Identify Cellular Receptors Used by Enteroviruses to Infect Cells
Anastasiia O. Sosnovtseva, Thi Hoa Le, Dmitry S. Karpov, Pavel O. Vorobyev, Yana D. Gumennaya, Olga N. Alekseeva, Peter M. Chumakov, Anastasia V. Lipatova
International Journal of Molecular Sciences, 2025
Non-pathogenic natural and recombinant strains of human Enteroviruses are the subject of ongoing study with some strains having been approved for use as anticancer agents. The efficacy of oncolytic virotherapy depends upon identifying the receptor utilized by a specific strain for cell entry, and the presence of this receptor on the surface of cancer cells. Accordingly, a rapid and straightforward approach to determining the enteroviral receptors is necessary for developing an effective patient-specific, virus-based cancer therapy. To this end, we created a panel of seven lines with double knockouts on the background of the HEK293T cell line, which lacks the IFNAR1 gene. In these lines, the main viral receptor genes, including PVR, CXADR, CD55, ITGA2, SCARB2, ICAM1, and FCGRT, were knocked out using the CRISPR/Cas9 system. The panel of lines was validated on twelve different Enteroviruses types, providing a basis for studying the molecular mechanisms of enterovirus entry into cells, and for developing new therapeutic strains. - INTERFERON TYPE I-EXPRESSING RECOMBINANT VACCINIA VIRUS AS A PLATFORM FOR SELECTIVE IMMUNOTHERAPY OF GLIOBLASTOMA AND MELANOMA
ER Naberezhnaya, AV Soboleva, PO Vorobyev, VV Vadekhina, GM Yusubalieva, IV Isaeva, VP Baklaushev, PM Chumakov, AV Lipatova
Bulletin of Russian State Medical University, 2024
Immunotherapy with oncolytic viruses (OVs) becomes a full-fledged neoadjuvant therapy method in the paradigm of evidence-based medicine for the growing number of cancers. The use of OVs for immunologically “cold” tumors causing minimal immune response and having the clearly immunosuppressive tumor microenvironment is especially relevant. Recombinant OVs carrying the sequences of proteins activating the immune system can be used to stimulate antitumor response. The study aimed to assess oncoselectivity and antitumor activity of the recombinant OV designed based on the LIVP vaccinia virus strain showing expression of human and murine interpheron alpha sequences (hIFNα and mIFNα, respectively). The in vitro experiments showed that the recombinant OVs designed showed oncoselectivity in relation to tumor cell lines of appropriate species. The ability to effectively infect human adenocarcinoma and glioblastoma cell lines was reported for LIVP-hIFNα. LIVP-mIFNα showed selectivity in relation to glioma Gl261 and melanoma B16 in vitro. The in vivo experiment involving the C57Bl/6 mice with subcutaneous melanoma В16 showed the ability of the intravenously administered LIVP-mIFNα to reduce the size of the subcutaneous tumor allograft and increase tumor infiltration with the CD8+ and NK cells. The recombinant virus designed can be a potential platform for the development of oncolytic virotherapy of human melanoma and glioblastoma. - A Snapshot of Early Transcriptional Changes Accompanying the Pro-Neural Phenotype Switch by NGN2, ASCL1, SOX2, and MSI1 in Human Fibroblasts: An RNA-Seq Study
Ekaterina M. Samoilova, Daria A. Chudakova, Erdem B. Dashinimaev, Anastasiya V. Snezhkina, Olga M. Kudryashova, Anastasia V. Lipatova, Alesya V. Soboleva, Pavel O. Vorob’yev, Vladimir T. Valuev-Elliston, Natalia F. Zakirova, Alexander V. Ivanov, Vladimir P. Baklaushev
International Journal of Molecular Sciences, 2024
Direct pro-neural reprogramming is a conversion of differentiated somatic cells to neural cells without an intermediate pluripotency stage. It is usually achieved via ectopic expression (EE) of certain transcription factors (TFs) or other reprogramming factors (RFs). Determining the transcriptional changes (TCs) caused by particular RFs in a given cell line enables an informed approach to reprogramming initiation. Here, we characterized TCs in the human fibroblast cell line LF1 on the 5th day after EE of the single well-known pro-neural RFs NGN2, ASCL1, SOX2, and MSI1. As assessed by expression analysis of the bona fide neuronal markers nestin and beta-III tubulin, all four RFs initiated pro-neuronal phenotype conversion; analysis by RNA-seq revealed striking differences in the resulting TCs, although some pathways were overlapping. ASCL1 and SOX2 were not sufficient to induce significant pro-neural phenotype switches using our EE system. NGN2 induced TCs indicative of cell phenotype changes towards neural crest cells, neural stem cells, mature neurons, as well as radial glia, astrocytes, and oligodendrocyte precursors and their mature forms. MSI1 mainly induced a switch towards early stem-like cells, such as radial glia. - Receptors and Host Factors for Enterovirus Infection: Implications for Cancer Therapy
Olga N. Alekseeva, Le T. Hoa, Pavel O. Vorobyev, Dmitriy V. Kochetkov, Yana D. Gumennaya, Elizaveta R. Naberezhnaya, Denis O. Chuvashov, Alexander V. Ivanov, Peter M. Chumakov, Anastasia V. Lipatova
Cancers, 2024
Enteroviruses, with their diverse clinical manifestations ranging from mild or asymptomatic infections to severe diseases such as poliomyelitis and viral myocarditis, present a public health threat. However, they can also be used as oncolytic agents. This review shows the intricate relationship between enteroviruses and host cell factors. Enteroviruses utilize specific receptors and coreceptors for cell entry that are critical for infection and subsequent viral replication. These receptors, many of which are glycoproteins, facilitate virus binding, capsid destabilization, and internalization into cells, and their expression defines virus tropism towards various types of cells. Since enteroviruses can exploit different receptors, they have high oncolytic potential for personalized cancer therapy, as exemplified by the antitumor activity of certain enterovirus strains including the bioselected non-pathogenic Echovirus type 7/Rigvir, approved for melanoma treatment. Dissecting the roles of individual receptors in the entry of enteroviruses can provide valuable insights into their potential in cancer therapy. This review discusses the application of gene-targeting techniques such as CRISPR/Cas9 technology to investigate the impact of the loss of a particular receptor on the attachment of the virus and its subsequent internalization. It also summarizes the data on their expression in various types of cancer. By understanding how enteroviruses interact with specific cellular receptors, researchers can develop more effective regimens of treatment, offering hope for more targeted and efficient therapeutic strategies. - Polyamine Catabolism Revisited: Acetylpolyamine Oxidase Plays a Minor Role Due to Low Expression
Olga N. Ivanova, Anna V. Gavlina, Inna L. Karpenko, Martin A. Zenov, Svetlana S. Antseva, Natalia F. Zakirova, Vladimir T. Valuev-Elliston, George S. Krasnov, Irina T. Fedyakina, Pavel O. Vorobyev, Birke Bartosch, Sergey N. Kochetkov, Anastasiya V. Lipatova, Dmitry V. Yanvarev, Alexander V. Ivanov
Cells, 2024
Biogenic polyamines are ubiquitous compounds. Dysregulation of their metabolism is associated with the development of various pathologies, including cancer, hyperproliferative diseases, and infections. The canonical pathway of polyamine catabolism includes acetylation of spermine and spermidine and subsequent acetylpolyamine oxidase (PAOX)-mediated oxidation of acetylpolyamines (back-conversion) or their direct efflux from the cell. PAOX is considered to catalyze a non-rate-limiting catabolic step. Here, we show that PAOX transcription levels are extremely low in various tumor- and non-tumor cell lines and, in most cases, do not change in response to altered polyamine metabolism. Its enzymatic activity is undetectable in the majority of cell lines except for neuroblastoma and low passage glioblastoma cell lines. Treatment of A549 cells with N1,N11-diethylnorspermine leads to PAOX induction, but its contribution to polyamine catabolism remains moderate. We also describe two alternative enzyme isoforms and show that isoform 4 has diminished oxidase activity and isoform 2 is inactive. PAOX overexpression correlates with the resistance of cancer cells to genotoxic antitumor drugs, indicating that PAOX may be a useful therapeutic target. Finally, PAOX is dispensable for the replication of various viruses. These data suggest that a decrease in polyamine levels is achieved predominantly by the secretion of acetylated spermine and spermidine rather than by back-conversion. - Preclinical Safety Assessment of Antineoplastic Agents Based on Live Non-Pathogenic Enteroviruses
K. V. Sivak, K. I. Stosman, E. Yu. Kalinina, M. M. Lyubishin, Ya. R. Orshanskaya, T. N. Savateeva-Lyubimova, S. B. Kazakova, D. M. Fedoseeva, A. S. Nazarov, E. I. Radion, S. M. Yudin, A. A. Keskinov, V. V. Makarov, V. S. Yudin, A. O. Zheltukhin, P. O. Vorobyev, D. V. Kochetkov, G. V. Ilyinskaya, A. V. Lipatova, P. M. Chumakov
Safety and Risk of Pharmacotherapy, 2024
INTRODUCTION. Developing novel medicines based on non-pathogenic enterovirus strains exhibiting oncotropic and oncolytic properties represents an up-to-date and safe approach to complex cancer treatment and postoperative metastasis prevention. Safety pharmacology studies are a necessary step in the preclinical development of medicinal products.AIM. The study aimed to investigate the single and repeated-dose general toxicity, local tolerance, safety pharmacology, and pyrogenicity of medicinal products based on non-pathogenic LEV4, LEV7, LEV8, LEV14, and Russo enterovirus strains as part of preclinical safety studies.MATERIALS AND METHODS. The study used medicinal products of highly purified group A, B, and C enteroviruses at a titre of 2×107–5×108 CPD50/mL (CPD50 is a cytopathogenic dose of the virus causing 50% cell lysis) and normal saline as a diluent. The viruses were propagated in Vero cells. The safety study used 220 male and female BALB/c mice, 440 male and female Wistar rats, and 18 male Soviet chinchilla rabbits. The study animals received an intravenous dose of 1×105 or 1×106 CPD50/animal once (single-dose toxicity) or weekly for 90 days (repeated-dose toxicity). Clinical examination, laboratory testing, and necropsy were performed on Days 45 and 91 of the experiment. Statistical data processing was performed using Prism 8.0 software (GraphPad Software, Inc., USA).RESULTS. Upon single administration of each of the five enterovirus medicinal products to mice and rats, the authors observed complete survival, upward trends in body weight gain, and no gross or histopathological changes in the brain, spleen, liver, kidneys, lungs, or at the injection site. Upon repeated administration at the study doses, the medicinal products caused no functional changes in the organs and systems. All the studied parameters were within the normal physiological ranges for male and female rats. Histopathological examination revealed no pathological changes or specific cytolytic and/or cytopathic effects. No local irritation was observed. None of the investigational medicinal products showed pyrogenicity.CONCLUSIONS. The obtained preclinical results demonstrate the safety of antineoplastic agents based on live non-pathogenic LEV4, LEV7, LEV8, LEV14, and Russo enteroviruses. - Oncolytic therapy with recombinant vaccinia viruses targeting the interleukin-15 pathway elicits a synergistic response
Yasmin Shakiba, Pavel O. Vorobyev, Gaukhar M. Yusubalieva, Dmitry V. Kochetkov, Ksenia V. Zajtseva, Marat P. Valikhov, Vladimir A. Kalsin, Fedor G. Zabozlaev, Alevtina S. Semkina, Alexander V. Troitskiy, Vladimir P. Baklaushev, Peter M. Chumakov, Anastasia V. Lipatova
Molecular Therapy Oncolytics, 2023
We developed recombinant variants of oncolytic vaccinia virus LIVP strain expressing interleukin-15 (IL-15) or its receptor subunit alpha (IL-15Rα) to stimulate IL-15-dependent immune cells. We evaluated their oncolytic activity either alone or in combination with each other in vitro and in vivo using the murine CT26 colon carcinoma and 4T1 breast carcinoma models. We demonstrated that the admixture of these recombinant variants could promote the generation of the IL-15/IL-15Rα complex. In vitro studies indicated that 4T1 breast cancer cells were more susceptible to the developed recombinant viruses. In vivo studies showed significant survival benefits and tumor regression in 4T1 breast cancer syngeneic mice that received a combination of LIVP-IL15-RFP with LIVP-IL15Ra-RFP. Histological analysis showed recruited lymphocytes at the tumor region, while no harmful effects to the liver or spleen of the animals were detected. Evaluating tumor-infiltrated lymphocytes represented profound activation of cytotoxic T cells and macrophages in mice receiving combination therapy. Thus, our experiments showed superior oncolytic effectiveness of simultaneous injection of LIVP-IL15-RFP and LIVP-IL15Ra-RFP in breast cancer-bearing mice. The combined therapy by these recombinant variants represents a potent and versatile approach for developing new immunotherapies for breast cancer. We developed recombinant variants of oncolytic vaccinia virus LIVP strain expressing interleukin-15 (IL-15) or its receptor subunit alpha (IL-15Rα) to stimulate IL-15-dependent immune cells. We evaluated their oncolytic activity either alone or in combination with each other in vitro and in vivo using the murine CT26 colon carcinoma and 4T1 breast carcinoma models. We demonstrated that the admixture of these recombinant variants could promote the generation of the IL-15/IL-15Rα complex. In vitro studies indicated that 4T1 breast cancer cells were more susceptible to the developed recombinant viruses. In vivo studies showed significant survival benefits and tumor regression in 4T1 breast cancer syngeneic mice that received a combination of LIVP-IL15-RFP with LIVP-IL15Ra-RFP. Histological analysis showed recruited lymphocytes at the tumor region, while no harmful effects to the liver or spleen of the animals were detected. Evaluating tumor-infiltrated lymphocytes represented profound activation of cytotoxic T cells and macrophages in mice receiving combination therapy. Thus, our experiments showed superior oncolytic effectiveness of simultaneous injection of LIVP-IL15-RFP and LIVP-IL15Ra-RFP in breast cancer-bearing mice. The combined therapy by these recombinant variants represents a potent and versatile approach for developing new immunotherapies for breast cancer. - Recombinant Strains of Oncolytic Vaccinia Virus for Cancer Immunotherapy
Yasmin Shakiba, Pavel O. Vorobyev, Marah Mahmoud, Azzam Hamad, Dmitriy V. Kochetkov, Gaukhar M. Yusubalieva, Vladimir P. Baklaushev, Peter M. Chumakov, Anastasia V. Lipatova
Biochemistry Moscow, 2023
Cancer virotherapy is an alternative therapeutic approach based on the viruses that selectively infect and kill tumor cells. Vaccinia virus (VV) is a member of the Poxviridae, a family of enveloped viruses with a large linear double-stranded DNA genome. The proven safety of the VV strains as well as considerable transgene capacity of the viral genome, make VV an excellent platform for creating recombinant oncolytic viruses for cancer therapy. Furthermore, various genetic modifications can increase tumor selectivity and therapeutic efficacy of VV by arming it with the immune-modulatory genes or proapoptotic molecules, boosting the host immune system, and increasing cross-priming recognition of the tumor cells by T-cells or NK cells. In this review, we summarized the data on bioengineering approaches to develop recombinant VV strains for enhanced cancer immunotherapy. - Oncolytic Efficacy of a Recombinant Vaccinia Virus Strain Expressing Bacterial Flagellin in Solid Tumor Models
Yasmin Shakiba, Pavel O. Vorobyev, Victor A. Naumenko, Dmitry V. Kochetkov, Ksenia V. Zajtseva, Marat P. Valikhov, Gaukhar M. Yusubalieva, Yana D. Gumennaya, Egor A. Emelyanov, Alevtina S. Semkina, Vladimir P. Baklaushev, Peter M. Chumakov, Anastasia V. Lipatova
Viruses, 2023
Oncolytic viral therapy is a promising novel approach to cancer treatment. Oncolytic viruses cause tumor regression through direct cytolysis on the one hand and recruiting and activating immune cells on the other. In this study, to enhance the antitumor efficacy of the thymidine kinase-deficient vaccinia virus (VV, Lister strain), recombinant variants encoding bacterial flagellin (subunit B) of Vibrio vulnificus (LIVP-FlaB-RFP), firefly luciferase (LIVP-Fluc-RFP) or red fluorescent protein (LIVP-RFP) were developed. The LIVP-FLuc-RFP strain demonstrated exceptional onco-specificity in tumor-bearing mice, detected by the in vivo imaging system (IVIS). The antitumor efficacy of these variants was explored in syngeneic murine tumor models (B16 melanoma, CT26 colon cancer and 4T1 breast cancer). After intravenous treatment with LIVP-FlaB-RFP or LIVP-RFP, all mice tumor models exhibited tumor regression, with a prolonged survival rate in comparison with the control mice. However, superior oncolytic activity was observed in the B16 melanoma models treated with LIVP-FlaB-RFP. Tumor-infiltrated lymphocytes and the cytokine analysis of the serum and tumor samples from the melanoma-xenografted mice treated with these virus variants demonstrated activation of the host’s immune response. Thus, the expression of bacterial flagellin by VV can enhance its oncolytic efficacy against immunosuppressive solid tumors. - COMPARISON OF THE ONCOLYTIC ACTIVITY OF RECOMBINANT VACCINIA VIRUS STRAINS LIVP-RFP AND MVA-RFP AGAINST SOLID TUMORS
Y Shakiba, ER Naberezhnaya, DV Kochetkov, GM Yusubalieva, PO Vorobyev, PM Chumakov, VP Baklaushev, AV Lipatova
Bulletin of Russian State Medical University, 2023 - SARS-CoV-2 Establishes a Productive Infection in Hepatoma and Glioblastoma Multiforme Cell Lines
Olga A. Smirnova, Olga N. Ivanova, Irina T. Fedyakina, Gaukhar M. Yusubalieva, Vladimir P. Baklaushev, Dmitry V. Yanvarev, Olga I. Kechko, Vladimir A. Mitkevich, Pavel O. Vorobyev, Vyacheslav S. Fedorov, Birke Bartosch, Vladimir T. Valuev-Elliston, Anastasiya L. Lipatova, Alexander V. Ivanov
Cancers, 2023 - SINGLE-DOMAIN ANTIBODY FOR BINDING THE CONSERVED EPITOPE IN THE SARS-COV-2 SPIKE PROTEIN RECEPTOR-BINDING DOMAIN
PO Vorobyev, SV Tillib
Bulletin of Russian State Medical University, 2023 - 2-Deoxyglucose, an Inhibitor of Glycolysis, Enhances the Oncolytic Effect of Coxsackievirus
Pavel O. Vorobyev, Dmitry V. Kochetkov, Peter M. Chumakov, Natalia F. Zakirova, Sofia I. Zotova-Nefedorova, Konstantin V. Vasilenko, Olga N. Alekseeva, Sergey N. Kochetkov, Birke Bartosch, Anastasiya V. Lipatova, Alexander V. Ivanov
Cancers, 2022 - Oncolytic Viruses in the Therapy of Lymphoproliferative Diseases
P. O. Vorobyev, F. E. Babaeva, A. V. Panova, J. Shakiba, S. K. Kravchenko, A. V. Soboleva, A. V. Lipatova
Molecular Biology, 2022 - Oncolytic Viruses in the Therapy of Lymphoproliferative Diseases
Pavel O. Vorobyev, F E Babaeva, Alexandra V. Panova, J. Shakiba, S K Kravchenko, et al.
Molekuliarnaia Biologiia, 2022 - COMPARATIVE EFFICIENCY OF ACCESSIBLE TRANSFECTION METHODS IN MODEL CELL LINES FOR BIOTECHNOLOGICAL APPLICATIONS
PO Vorobyev, DV Kochetkov, KV Vasilenko, AV Lipatova
Bulletin of Russian State Medical University, 2022 - Multiomic Profiling Identified EGF Receptor Signaling as a Potential Inhibitor of Type I Interferon Response in Models of Oncolytic Therapy by Vesicular Stomatitis Virus
Anastasia S. Nikitina, Anastasia V. Lipatova, Anton O. Goncharov, Anna A. Kliuchnikova, Mikhail A. Pyatnitskiy, Ksenia G. Kuznetsova, Azzam Hamad, Pavel O. Vorobyev, Olga N. Alekseeva, Marah Mahmoud, Yasmin Shakiba, Ksenia S. Anufrieva, Georgy P. Arapidi, Mark V. Ivanov, Irina A. Tarasova, Mikhail V. Gorshkov, Peter M. Chumakov, Sergei A. Moshkovskii
International Journal of Molecular Sciences, 2022 - DEVELOPMENT OF A RECOMBINANT ONCOLYTIC POLIOVIRUS TYPE 3 STRAIN WITH ALTERED CELL TROPISM
A Hamad, AV Soboleva, PO Vorobyev, M Mahmoud, KV Vasilenko, PM Chumakov, AV Lipatova
Bulletin of Russian State Medical University, 2022 - Study of the Therapeutic Efficiency of Transduced Olfactory Ensheathing Cells in Spinal Cord Cysts
Olga V. Stepanova, Anastasia D. Voronova, Anastasiia O. Sosnovtseva, Aleksei A. Stepanenko, Andrey V. Chadin, Ekaterina K. Karsuntseva, Grigorii A. Fursa, Marat P. Valikhov, Alevtina S. Semkina, Pavel O. Vorobyev, Igor V. Reshetov, Vladimir P. Chekhonin
Stem Cells and Development, 2022 - Multi-omics analysis of glioblastoma cells’ sensitivity to oncolytic viruses
Anastasiya V. Lipatova, Alesya V. Soboleva, Vladimir A. Gorshkov, Julia A. Bubis, Elizaveta M. Solovyeva, George S. Krasnov, Dmitry V. Kochetkov, Pavel O. Vorobyev, Irina Y. Ilina, Sergei A. Moshkovskii, Frank Kjeldsen, Mikhail V. Gorshkov, Peter M. Chumakov, Irina A. Tarasova
Cancers, 2021 - Effects of Siberian fir terpenes extract Abisil on antioxidant activity, autophagy, transcriptome and proteome of human fibroblasts
Anastasiya Lipatova, George Krasnov, Pavel Vorobyov, Pavel Melnikov, Olga Alekseeva, Yulia Vershinina, Alexander Brzhozovskiy, Daria Goliusova, Faniya Maganova, Natalia Zakirova, Anna Kudryavtseva, Alexey Moskalev
Aging, 2021
