Roman N. Chuprov‐Netochin

Moscow Institute of Physics and Technology

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Sergey Leonov

Institute of Cell Biophysics

Dmitry A. Gorin

Skolkovo Institute of Science and Technology

Marina N. Semenova

Koltzov Institute of Developmental Biology

Vasiliy S. Chernyshev

National Medical Research Center for Obstetrics, Gynecology and Perinatology named after Academician V.I.Kulakov of the Ministry of Healthcare of the Russian Federation

МП

Маргарита Пустовалова

Institute of Biophysics

Elena Marusich

Institute of Biophysics

Victor V. Semenov

N.D. Zelinsky Institute of Organic Chemistry

АН

А. Н. Осипов

State Scientific Center of the Russian Federation - Federal Medical Biophysical Center named after A.I. Burnazyan

Alexey M. Yashchenok

Skolkovo Institute of Science and Technology

Lina Alhaddad

Damascus University

Cooperative Institutions

Russian Academy of Sciences

Moscow Institute of Physics and Technology

Lomonosov Moscow State University

Skolkovo Institute of Science and Technology

Institute of Bioorganic Chemistry

Kurchatov Institute

Moscow Power Engineering Institute

N.D. Zelinsky Institute of Organic Chemistry

Ministry of Health of the Russian Federation

Moscow Aviation Institute

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IR-Surviving NSCLC Cells Exhibit Different Patterns of Molecular and Cellular Reactions Relating to the Multifraction Irradiation Regimen and p53-Family Proteins Expression

Cancers (2021)

Lina Alhaddad Маргарита Пустовалова Taisia Blokhina Roman N. Chuprov‐Netochin А. Н. Осипов

Radiotherapy is a primary treatment modality for patients with unresectable non-small cell lung cancer (NSCLC). Tumor heterogeneity still poses the central question of cancer radioresistance, whether the presence of a particular cell population inside a tumor undergoing a selective outgrowth during radio- and chemotherapy give rise to metastasis and tumor recurrence. In this study, we examined the impact of two different multifraction X-ray radiation exposure (MFR) regimens, fraction dose escalation (FDE) in the split course and the conventional hypofractionation (HF), on the phenotypic and molecular signatures of four MFR-surviving NSCLC cell sublines derived from parental A549 (p53 wild-type) and H1299 (p53-null) cells, namely A549FR/A549HR, H1299FR/H1299HR cells. We demonstrate that sublines surviving different MFR regimens in a total dose of 60 Gy significantly diverge in their molecular traits related to irradiation regimen and p53 status. The observed changes regarding radiosensitivity, transformation, proliferation, metabolic activity, partial epithelial-to-mesenchymal transition (EMT) program activation and 1D confined migratory behavior (wound healing). For the first time, we demonstrated that MFR exposure led to the significant decrease in the expression of p63 and p73, the p53-family members, in p53null cells, which correlated with the increase in cell polyploidy. We could not find significant differences in FRA1 expression between parental cells and their sublines that survived after any MFR regimen regardless of p53 status. In our study, the FDE regimen probably causes partial EMT program activation in MFR-survived NSCLC cells through either Vimentin upregulation in p53null or an aberrant N-cadherin upregulation in p53wt cells. The HF regimen likely less influences the EMT activation irrespectively of the p53 status of MFR-survived NSCLC cells. Our data highlight that both MFR regimens caused overall higher cell transformation of p53null H1299FR and H1299HR cells than their parental H1299 cells. Moreover, our results indicate that the FDE regimen raised the radioresistance and transformation of MFR-surviving NSCLC cells irrespectively of their p53 status, though the HF regimen demonstrated a similar effect on p53null NSCLC cells only. Our data once again emphasize that NSCLC therapy approaches should become more personalized according to radiation therapy (RT) regimen, tumor histology, and molecular status of critical proteins.

Radioresistance

Regimen

Radiosensitivity

10.3390/cancers13112669

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Citations (11)

Synthesis of chalcones with methylenedioxypolymethoxy fragments based on plant metabolites and study of their antiproliferative properties

Russian Chemical Bulletin (2023)

Alexander E. Varakutin Еgor А. Muravsky Dmitry V. Tsyganov Ilia Yu Shinkarev Aida I. Samigullina

Chalcone

Conformational isomerism

Cancer cell lines

10.1007/s11172-023-3943-6

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Citations (5)

Gold nanoparticle-carbon nanotube multilayers on silica microspheres: Optoacoustic-Raman enhancement and potential biomedical applications

Materials Science and Engineering C (2020)

Daniil Nozdriukhin Nadezhda A. Besedina Vasiliy S. Chernyshev Olga Efimova Polina G. Rudakovskaya

Multiplex

Nanorod

10.1016/j.msec.2020.111736

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Citations (18)

Effect of Surface Modification of Multifunctional Nanocomposite Drug Delivery Carriers with DARPin on Their Biodistribution In Vitro and In Vivo

ACS Applied Bio Materials (2022)

Marina V. Novoselova Vasiliy S. Chernyshev Alexey Schulga Е. В. Коновалова Roman N. Chuprov‐Netochin

We present a targeted drug delivery system for therapy and diagnostics that is based on a combination of contrasting, cytotoxic, and cancer-cell-targeting properties of multifunctional carriers. The system uses multilayered polymer microcapsules loaded with magnetite and doxorubicin. Loading of magnetite nanoparticles into the polymer shell by freezing-induced loading (FIL) allowed the loading efficiency to be increased 5-fold, compared with the widely used layer-by-layer (LBL) assembly. FIL also improved the photoacoustic signal and particle mobility in a magnetic field gradient, a result unachievable by the LBL alone. For targeted delivery of the carriers to cancer cells, the carrier surface was modified with a designed ankyrin repeat protein (DARPin) directed toward the epithelial cell adhesion molecule (EpCAM). Flow cytometry measurements showed that the DARPin-coated capsules specifically interacted with the surface of EpCAM-overexpressing human cancer cells such as MCF7. In vivo and ex vivo biodistribution studies in FvB mice showed that the carrier surface modification with DARPin changed the biodistribution of the capsules toward epithelial cells. In particular, the capsules accumulated substantially in the lungs─a result that can be effectively used in targeted lung cancer therapy. The results of this work may aid in the further development of the "magic bullet" concept and may bring the quality of personalized medicine to another level.

Biodistribution

Surface Modification

Targeted drug delivery

10.1021/acsabm.2c00289

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Citations (6)

Engineered multicompartment vesicosomes for selective uptake by living cells

Colloids and Surfaces B Biointerfaces (2022)

Vasiliy S. Chernyshev Daniil Nozdriukhin Roman N. Chuprov‐Netochin Ekaterina Tsydenzhapova Marina V. Novoselova

Extracellular Vesicles

10.1016/j.colsurfb.2022.112953

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Citations (6)

СВОЙСТВА ПЕПТИДОГЛИКАНЛИЗИРУЮЩЕГО ФЕРМЕНТА БАКТЕРИОФАГА PMG1 PSEUDOMONAS AERUGINOSA

Биоорганическая химия (2011)

M. B. Burkal’tseva M. Ivanova Н Л Клячко Е С Кострюкова В. Н. Лазарев

Выделенный бактериофаг Pseudomonas aeruginosa имеет отличительные особенности в литическом инфекционном цикле и образует ореол (зону просветления) вокруг негативных колоний. Фаг был изучен с точки зрения потенциального использования в терапевтических фаговых препаратах и как источник пептидогликан- и липополисахариддеградирующих ферментов. Частичное секвенирование генома показало высокую степень его гомологии с известным умеренным бактериофагом D3. В геноме была идентифицирована открытая рамка считывания, кодирующая литическую трансгликозилазу. Этот фермент был получен в рекомбинантном виде, изучены его активность и субстратная специфичность.

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The CD44high Subpopulation of Multifraction Irradiation-Surviving NSCLC Cells Exhibits Partial EMT-Program Activation and DNA Damage Response Depending on Their p53 Status

International Journal of Molecular Sciences (2021)

Маргарита Пустовалова Lina Alhaddad Taisia Blokhina Н. М. Сметанина A. K. Chigasova

Ionizing radiation (IR) is used for patients diagnosed with unresectable non-small cell lung cancer (NSCLC). However, radiotherapy remains largely palliative due to the survival of specific cell subpopulations. In the present study, the sublines of NSCLC cells, A549IR (p53wt) and H1299IR (p53null) survived multifraction X-ray radiation exposure (MFR) at a total dose of 60 Gy were investigated three weeks after the MFR course. We compared radiosensitivity (colony formation), expression of epithelial-mesenchymal transition (EMT) markers, migration activity, autophagy, and HR-dependent DNA double-strand break (DSB) repair in the bulk and entire CD44high/CD166high CSC-like populations of both parental and MFR survived NSCLC cells. We demonstrated that the p53 status affected: the pattern of expression of N-cadherin, E-cadherin, Vimentin, witnessing the appearance of EMT-like phenotype of MFR-surviving sublines; 1D confined migratory behavior (wound healing); the capability of an irradiated cell to continue to divide and form a colony of NSCLC cells before and after MFR; influencing the CD44/CD166 expression level in MFR-surviving NSCLC cells after additional single irradiation. Our data further emphasize the impact of p53 status on the decay of γH2AX foci and the associated efficacy of the DSB repair in NSCLC cells survived after MFR. We revealed that Rad51 protein might play a principal role in MFR-surviving of p53 null NSCLC cells promoting DNA DSB repair by homologous recombination (HR) pathway. The proportion of Rad51 + cells elevated in CD44high/CD166high population in MFR-surviving p53wt and p53null sublines and their parental cells. The p53wt ensures DNA-PK-mediated DSB repair for both parental and MFR-surviving cells irrespectively of a subsequent additional single irradiation. Whereas in the absence of p53, a dose-dependent increase of DNA-PK-mediated non-homologous end joining (NHEJ) occurred as an early post-irradiation response is more intensive in the CSC-like population MFR-surviving H1299IR, compared to their parental H1299 cells. Our study strictly observed a significantly higher content of LC3 + cells in the CD44high/CD166high populations of p53wt MFR-surviving cells, which enriched the CSC-like cells in contrast to their p53null counterparts. The additional 2 Gy and 5 Gy X-ray exposure leads to the dose-dependent increase in the proportion of LC3 + cells in CD44high/CD166high population of both parental p53wt and p53null, but not MFR-surviving NSCLC sublines. Our data indicated that autophagy is not necessarily associated with CSC-like cells’ radiosensitivity, emphasizing that careful assessment of other milestone processes (such as senescence and autophagy-p53-Zeb1 axis) of primary radiation responses may provide new potential targets modulated for therapeutic benefit through radiosensitizing cancer cells while rescuing normal tissue. Our findings also shed light on the intricate crosstalk between autophagy and the p53-related EMT, by which MFR-surviving cells might obtain an invasive phenotype and metastatic potential.

Radiosensitivity

10.3390/ijms22052369

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Citations (16)

Опыт анализа биологической активности урсодезоксихолевой кислоты в контексте in silico прогноза профиля экспрессии генов

Генетика (2020)

Я. Б. Нескородов С. Г. Марданлы Roman N. Chuprov‐Netochin

10.31857/s0016675820100094

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Citations (1)

Translesion DNA Synthesis and Reinitiation of DNA Synthesis in Chemotherapy Resistance

Biochemistry (Moscow) (2020)

Evgeniy S. Shilkin Elizaveta O. Boldinova Anastasia D. Stolyarenko Р. И. Гончарова Roman N. Chuprov‐Netochin

Primase

DNA polymerase II

DNA clamp

DNA polymerase delta

10.1134/s0006297920080039

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Citations (10)

ASYMMETRIC DEPTH-FILTRATION – A VERSATILE AND SCALABLE APPROACH FOR ISOLATION AND PURIFICATION OF EXTRACELLULAR VESICLES

BIOTECHNOLOGY STATE OF THE ART AND PERSPECTIVES (2021)

Roman N. Chuprov‐Netochin E. V. Svirshchevskaya Р. А. Полтавцева Anastasiia Merdalimova Alexey M. Yashchenok

A new method that is based on asymmetric depth-filtration was developed and tested for isolation of extracellular vesicles (EVs) from biological fluids such as human plasma, urine, Wharton's jelly and cell culture. Results showed that such method can be a reliable alternative for EV research and their application in medicine.

Extracellular Vesicles

Filtration (mathematics)

Isolation

Extracellular vesicles

10.37747/2312-640x-2021-19-55-56

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Citations (0)