DSA Faculty
API
← к списку преподавателей

Князев Евгений Николаевич

Факультет биологии и биотехнологии

Профиль на hse.ru ↗ тел.: 15093
Публикаций
0
Языков
1
Наград
7
Конференций
9
Профиль Публикации (33) Курсы (11)

Профессиональные интересы

биохимияонкологиямодель кишечника человека in vitroбиоинформатикаперсонифицированная медицинабиология ракаклеточная биологиятрехмерные клеточные моделимикрофлюидикамикрофизиологические системымолекулярная биологияплацентарный барьер

Должности

  • Доцент — Факультет биологии и биотехнологии, Базовая кафедра Института биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова РАН
  • Заведующий лабораторией — Факультет биологии и биотехнологии, Лаборатория молекулярной физиологии

Био

  • · Начал работать в НИУ ВШЭ в 2019 году.
  • · Научно-педагогический стаж: 14 лет.

Образование

  • 2016 · Кандидат медицинских наук: специальность Онкология, тема диссертации: МикроРНК-маркеры для малоинвазивной диагностики рака предстательной железы
  • 2013 · Специалитет: Московский государственный университет им. М.В. Ломоносова, факультет: Факультет фундаментальной медицины, специальность «Лечебное дело», квалификация «Врач»

Опыт работы

  • · 2019 — по настоящее время: Декабрь : факультет биологии и биотехнологии, Национальный исследовательский университет «Высшая школа экономики», доцент, заведующий лабораторией молекулярной физиологии
  • · 2019 — по настоящее время: Май : лаборатория микрофлюидных технологий для биомедицины, ФГБУН Государственный Научный Центр Российской Федерации Институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова РАН, старший научный сотрудник
  • · 2013: Февраль сентябрь
  • · 2023: : ООО «Центр трансляционных технологий» (ООО Лаборатория «БиоКлиникум»), генеральный директор
  • · 2022: Ноябрь июнь
  • · 2023: : факультет Биомедицинская техника, Московский государственный технический университет им. Н.Э. Баумана, доцент
  • · 2021: Август февраль
  • · 2022: : Городская клиническая больница №15 им О.М. Филатова, врач клинической лабораторной диагностики
  • · 2019: Февраль август
  • · 2019: : Национальный медицинский исследовательский центр терапии и профилактической медицины Министерства здравоохранения Российской Федерации, старший научный сотрудник
  • · 2013: Сентябрь июнь
  • · 2015: : Государственный научный центр Российской Федерации — Институт медико-биологических проблем РАН, научный сотрудник
  • · 2012: Февраль январь
  • · 2013: : ООО НТЦ «Клеточные Технологии», лаборант-исследователь

Награды и поощрения

  • · Благодарность первого проректора НИУ ВШЭ (март 2023)
  • · Надбавка за академические успехи и вклад в научную репутацию НИУ ВШЭ (2024–2026)
  • · Надбавка за публикацию в журнале из Списка А (и приравненном к нему научном издании) (2025–2026, 2023–2024)
  • · Надбавка за публикацию в международном рецензируемом научном издании (2022–2023, 2021–2022)
  • · Лучший преподаватель — 2024
  • · Победитель Конкурса лучших русскоязычных научных и научно-популярных работ работников НИУ ВШЭ – 2023
  • · Группа высокого профессионального потенциала (кадровый резерв НИУ ВШЭ)Категория "Новые преподаватели" (2024–2025)

Гранты и проекты

  • — · на соискание учёной степени кандидата наук

Конференции (9)

Показать все
  • · 2024: Седьмой онлайн-семинар по математическому моделированию в области иммунологии (Москва). Доклад: Моделирование динамики SARS-CoV-2 в клеточных линиях
  • · 2024: Второй Саммит разработчиков лекарственных препаратов «Сириус.Биотех» (Федеральная территория «Сириус»). Доклад: Использование платформы «Человек-на-чипе» при разработке лекарственных препаратов
  • · 2024: IV Конгресс молодых учёных. Доклад: Использование платформы Homunculus для моделирования органов-на-чипе
  • · 2019: International Federation of Placenta Associations 2019 (IFPA2019); 8th Latin American Symposium on Maternal-Fetal Interaction and Placenta (VIII SLIMP), Buenos Aires, Argentina (Buenos Aires). Доклад: Placenta-on-a-chip model for assessing the transport and toxicity of xenobiotics in vitro
  • · 2019: International Federation of Placenta Associations 2019 (IFPA2019); 8th Latin American Symposium on Maternal-Fetal Interaction and Placenta (VIII SLIMP), Buenos Aires, Argentina (Buenos Aires). Доклад: Application of impedance spectroscopy for analysis of BeWo clone b30 human choriocarcinoma cell line
  • · 2019: CTR Annual Conference 2019 (Cambridge). Доклад: Placenta-on-a-chip model for the assessment of drug transport and toxicity
  • · 2019: 22st European Congress on Alternatives to Animal Testing, 19th Annual Congress of EUSAAT, Linz, Austria (Linz). Доклад: Placenta-on-a-chip model for assessing the transport and toxicity of xenobiotics in vitro
  • · 2019: Второй международный форум онкологии и радиологии (Москва). Доклад: Модель плаценты-на-чипе in vitro для оценки транспорта и токсичности химиотерапевтических препаратов
  • · 2019: V Всероссийская Конференция по молекулярной онкологии (Москва). Доклад: Модель плаценты-на-чипе in vitro для оценки транспорта и токсичности химиотерапевтических препаратов

Идентификаторы исследователя

Публикации (33)

Intracellular Transport of Ribosome-Inactivating Proteins Depends on Annexin 13

2020 · ARTICLE · en

In the present study, we assessed the role of annexin 13 membrane-binding protein (ANXA13) in the intracellular transport of vesicles containing type II ribosome-inactivating proteins (RIP-IIs). A modified human intestinal epithelial cell line HT29 was used, in which the expression of ANXA13 was significantly reduced. The cytotoxic effect of ricin and viscumin was evaluated by modification of 28S ribosome RNA. The observed differences in the activity of toxins on the parental and modified HT29 lines indicate that ANXA13 plays a different role in the intracellular transport of vesicles containing the RIP-IIs.

DOI ↗ PDF ↗

Relationship between the Expression Level of PSMD11 and Other Proteasome Proteins with the Activity of Ricin and Viscumin

2020 · ARTICLE · en

The role of proteasome proteins and proteins of the ERAD system in the cytotoxicity of type II ribosome-inactivating proteins ricin and viscumin was investigated. For this, the cell line of colorectal adenocarcinoma HT29, as well as the HT29-sh002 line obtained on its basis, were used. On the basis on the proteome analysis of these lines and the estimation of the proportion of inactivated ribosomes, it was shown that the contribution of the proteasome to the degradation of the catalytic subunits of toxins is different. The role of the Cdc37 co-chaperone in maintaining the stability of A subunit of viscumin in the cytoplasm is shown.

DOI ↗ PDF ↗

Towards embedding Caco-2 model of gut interface in a microfluidic device to enable multi-organ models for systems biology

2019 · ARTICLE · en

A cancer cell line originating from human epithelial colorectal adenocarcinoma (Caco-2 cells) serves as a high capacity model for a preclinical screening of drugs. Recent need for incorporating barrier tissue into multi-organ chips calls for inclusion of Caco-2 cells into microperfused environment. This article describes a series of systems biology insights obtained from comparing Caco-2 models cells grown as conventional 2D layer and in a microfluidic chip. When basic electrical parameters of Caco-2 monolayers were evaluated using impedance spectrometry and MTT assays, no differences were noted. On the other hand, the microarray profiling of mRNAs and miRNAs revealed that grows on a microfluidic chip leads to the change in the production of specific miRNA, which regulate a set of genes for cell adhesion molecules (CAMs), and provide for more complete differentiation of Caco-2 monolayer. Moreover, the sets of miRNAs secreted at the apical surface of Caco-2 monolayers grown in conventional 2D culture and in microfluidic device differ. When integrated into a multi-tissue platform, Caco-2 cells may aid in generating insights into complex pathophysiological processes, not possible to dissect in conventional cultures.

DOI ↗ PDF ↗

Metabolic Reprogramming of Trophoblast Cells in Response to Hypoxia

2019 · ARTICLE · en

Hypoxia of trophoblast cells is an important regulator of normal development of the placenta. However, some pathological states associated with hypoxia, e.g. preeclampsia, impair the functions of placental cells. Oxyquinoline derivative inhibits HIF-prolyl hydroxylase by stabilizing HIF-1 transcription complex, thus modeling cell response to hypoxia. In human choriocarcinoma cells BeWo b30 (trophoblast model), oxyquinoline increased the expression of a core hypoxia response genes along with up-regulation of NOS3, PDK1, and BNIP3 genes and down-regulation of the PPARGC1B gene. These changes in the expression profile attest to activation of the metabolic cell reprogramming mechanisms aimed at reducing oxygen consumption by enabling the switch from aerobic to anaerobic glucose metabolism and the respective decrease in number of mitochondria. The possibility of practical use of the therapeutic properties of oxyquinoline derivatives is discussed.

DOI ↗ PDF ↗

Oxyquinoline-Dependent Changes in Claudin-Encoding Genes Contribute to Impairment of the Barrier Function of the Trophoblast Monolayer

2019 · ARTICLE · en

Natural response to hypoxia critically depends on rapid stabilization of hypoxia-inducible factor (HIF). Under normoxic conditions, HIF-prolyl hydroxylases mark α-subunits of HIF for degradation, while hypoxia results in stabilization of HIF-α. Oxyquinoline derivatives suppress activity of HIF-prolyl hydroxylases leading to HIF activation in the cell. Here we show that 24-h incubation of BeWo b30 choriocarcinoma cells (a model of trophoblast in the placental barrier) with oxyquinoline derivative leads to a decrease in transepithelial electrical resistance (TEER) of the cell monolayer, while the permeability of the monolayer for FITC-dextran (70 kDa) remains unchanged. These findings suggest that the overall barrier function is preserved, while the structure of intercellular tight junctions can undergo minor changes. Using Affymetrix Human Transcriptome Array 2.0, we showed that the treatment with oxyquinoline derivative was followed by a decrease in the expression of claudins 6 and 7 (CLDN6, CLDN7), occludin (OCLN), contact adhesion molecule 3 (JAM3), and angiomotinlike protein 1 (AMOTL1).

DOI ↗ PDF ↗

Detection of Low-Abundant MicroRNAs with Hybridization Microchips

2019 · ARTICLE · en

The effect of low concentrations of miRNA on the ability of GeneChip miRNA 4.0 hybridization chips to evaluate their representation in the sample was studied. It is shown that the evaluation of the expression of 61 miRNAs is statistically significantly associated with the multiplicity of plasma dilution. Only 12 miRNAs showed very high Pearson correlation coefficient (>0.95) and they all decreased in response to dilution. High abundance of has-miR-4532 miRNA in plasma was demonstrated. This miRNA was never detected during sequencing of similar samples. It was concluded that in case of miRNA expression

DOI ↗ PDF ↗

Transport and toxicity of 5-fluorouracil, doxorubicin, and cyclophosphamide in in vitro placental barrier model based on BeWo b30 cells

2019 · ARTICLE · en

An in vitro placental barrier model based on human choriocarcinoma BeWo b30 cell line was considered as a method of preclinical study of the transport and toxicity of antitumor agents and other organic compounds. Low permeabilities were found for 5-fluorouracil as an example of hydrophilic compound and for doxorubicin as an example of a lipophilic compound with a high degree of binding to proteins and DNA and a high permeability was found for cyclophosphamide as an example of lipophilic compound with a low degree of binding to proteins. Using impedance spectrometry and cell viability assessment via reduction of resazurin to resorufin, a pronounced cytotoxic effect of doxorubicin and good tolerance of 5-fluorouracil and cyclophosphamide by the cells were shown for drug concentrations equal to the maximum concentrations in the patients’ blood during the treatment of breast cancer.

DOI ↗ PDF ↗

Application of impedance spectroscopy for analysis of BeWo clone b30 human choriocarcinoma cell line

2019 · ARTICLE · en

Objectives: BeWo cells are used for the construction of in vitro models of placental barrier. Only the early placenta possesses multiple trophoblast layers whereas third-trimester placenta consists of a single trophoblast layer. BeWo cells do not undergo contact growth inhibition and form multilayer structures. It is important to control the cell state for transport experiments. Impedance spectroscopy was applied for electrical characteristic studying during BeWo cell growth and in response to HIF-1 activator. Methods: 30,000 cells per insert were seeded into a 96-well Transwell plate (1 μm pore size). After 48 h a potent HIF-1 activator D014-0021 was added at 10 μM concentration. Impedance spectra were acquired with impedance spectroscopy system (Bioclinicum, Russia). For the extraction of electrical parameters, the equivalent electrical circuits were used. Student's t-test was used to calculate the statistical significance. Results: It was predicted from the mathematical model that medium resistance (Rmed) and the radius of the impedance hodograph (and hence TEER) will rise linearly with the number of layers. In the case of one layer, the radius can rise due to the formation of tight junctions but the Rmed should remain stable, but the data shows increasing TEER and Rmed. There is a statistically significant difference in Rmed between 48 and 96 h. It can be concluded that after 48 h the BeWo cells form multilayer structures. The addition of D014-0021 leads to a slight increase in TEER after 6 h and a significant decrease in TEER and capacitance after 27 h. Conclusion: It was shown that formation of multilayer structures can be readily detected with impedance spectroscopy and hence it can be used for quality control of the in vitro placental models. It is possible to use impedance spectroscopy for detection of additional parameter changes such as electrical capacitance. This work is supported by the Russian Science Foundation (project No. 16-19-10597).

DOI ↗

Placenta-on-a-chip model for assessing the transport and toxicity of xenobiotics in vitro

2019 · ARTICLE · en

Objectives: The study of the transport and toxicity of xenobiotics in women is limited for ethical reasons. Ex vivo placenta models have high variability and low success rates. Animal models in vivo differ from a human in anatomy, genotype, and proteome. The placenta-on-a-chip model is a compromise. We studied the components of the FAC chemotherapy regimen for breast cancer in this model. Methods: BeWo b30 cell line was grown in the DMEM with L-glutamine, 4.5 g glucose/l and Earle’s salts containing 10% FBS, 1x MEM NEAA, 100 U/ml penicillin and 100 μg/ml streptomycin in inserts cut from 96-well Transwell plate and placed in a microfluidic chip. Cells were seeded with a density of 10,000 cells per insert. After 7 days, 5-fluorouracil (25 μg/ml), doxorubicin (50 μg/ml), cyclophosphamide (150 μg/ml), or all three drugs were added to the cells for 1 hour. Control cells were cultured in the presence of 0.05% DMSO. The impedance spectrum was measured before and 1 and 24 hours after the addition of the drug. The concentration of the drug was determined by HPLC-MS/MS. Cell viability was assessed using the CellTiter-Blue Assay. Results: After 1 h incubation with drugs, TEER decreased in experiment and control groups from an average of 90 to 25 Ω∙cm2, and after 24 h TEER was 67.3±17.9 Ω∙cm2 for control, 67.8±16.4 Ω∙cm2 for cyclophosphamide, 90.0±20.1 Ω∙cm2 for 5-fluorouracil, and decreased to the background for doxorubicin and drug mixture. Cell viability did not differ significantly between the control, 5-fluorouracil, and cyclophosphamide groups, but decreased to 40±9% of the control when exposed to doxorubicin and drug mixture. The placenta-on-a-chip model transported the drugs from the apical to the basolateral side. Conclusion: The developed placenta-on-a-chip model is suitable for assessing the transport and toxicity of xenobiotics in vitro. This work is supported by the Ministry of Science and Higher Education of the Russian Federation, project: 14.588.21.0007, unique id: RFMEFI58817X0007.

DOI ↗

Метаболическое перепрограммирование трофобласта при моделировании ответа на гипоксию

2018 · ARTICLE · ru

Гипоксия клеток трофобласта является важным регулирующим фактором в процессе нормального развития плаценты. Однако воздействие гипоксии на плаценту при ряде патологических состояний, таких как преэклампсия, приводит к нарушению функций клеток. Производное оксихинолина способно ингибировать HIF-пролилгидроксилазы, стабилизируя таким образом транскрипционный комплекс HIF-1 и моделируя ответ клетки на гипоксию. Клетки хориокарциномы человека BeWo b30 используют для моделирования трофобласта, который является основой плацентарного барьера. При воздействии оксихинолина было выявлено не только повышение экспрессии целого ряда генов “ядра ответа” на гипоксию, но и повышение экспрессии генов NOS3, PDK1, BNIP3 и снижение экспрессии гена PPARGC1B. Это указывает на активацию механизмов метаболического перепрограммирования клеток, направленного на снижение потребления кислорода за счет уменьшения числа митохондрий и перехода от аэробного метаболизма глюкозы к анаэробному. Обсуждается возможность практического применения полученных результатов.

DOI ↗ PDF ↗

Курсы (11)