Preclinical acute toxicity studies of the CAR technology products for malignant neoplasms therapy on the example of the «anti-HER2-CAR-T/CAR-NK»

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Abstract

Introduction. The Car-T-cell therapy is current and modern promising method for the oncology treatment. USA Food and Drug Sanitary Control Department confirms six drugs for cellular immunotherapy the blood oncology today. But the information about biomedical product preclinical test is absent, because used T-cells (cell with chimeric antigen receptor) are autologous for humans, which is the problem for use classic toxicity tests. Besides the biomedical product safety becomes questionable, therefore the different preclinical research strategy is developed to solve those problems and produce normal test-systems with therapeutic target.

The aim of the study was to evaluate the acute toxicity in immunodeficient BALB/c Nude mice of an antineoplastic drug based on genetically modified T/NK cells that express a chimeric T-cell anti-HER2 receptor.

Material and methods. Test and control groups consisted of five males and five females. Animals were injected a single intravenous or intraperitoneal injection of the testing product and the solvent-cryopreserving carrier at the dose 0,2 ml/animal. There were two doses: equal to the human therapeutic 0,5•106 cell/animal dose and ten times over then the therapeutic dose 5•106 cell/animal. During the test the animal’s weight, the food intake and clinically symptoms of the testing product toxicity were registered. On the fifteenth of the study day animals were euthanatized and exposed to a necropsy with the organs’ macroscopic inspection, the weighting and fixating. The detection of the testing biomedical product toxicity was the aim of the histology analysis.

Results. A single intravenous or intraperitoneal injection of the biomedical product «anti-HER2-CAR-T/ CAR-NK» at the human therapeutic dose, as well as in 10 times more than the same, is safe for BALB/c Nude mice. The preclinical study has shown the absence of significant toxic effects.

Limitations. The research was performed on Balb/c nude line immunodeficient mice, because the tested product contained living foreign cells.

Conclusion. This work can be the main basis for the creation of biomedical product preclinical research protocol of biomedical cell products produced from CAR-technology.

Compliance with ethical standards. All manipulations with animals were carried out in accordance with the Animal Application Protocol, which was examined and approved of IACUC of the Branch of the IBCh RAS.

Author contribution:
Goryacheva N.А., Bondarenko D.А., Rzhevskiy D.I., Slashheva G.A., Novikova N.I., Dyachenko I.A., Murashev A.N. — сollection and processing of material, literature review, writing the text;
Rzhevskiy D.I. — сollection and processing of material, statistical analysis;
Kisilevskiy M.V., Chikileva I.O., Vlasenko R.Ya. — the concept and design of the study.
All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version.

Conflict of interests. The authors declare no conflict of interests.

Funding. The study had no sponsorship.

Received: February 08, 2022 / Accepted: December 12, 2022 / Published: December 30, 2022

About the authors

Natal’ya Aleksandrovna Goryacheva

Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences; Pushchino State Institute of Natural Sciences

Author for correspondence.
Email: natasha.goryacheva2017@yandex.ru
ORCID iD: 0000-0003-4004-5214

Junior researcher of Biological Testing Laboratory, Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences, PhD student of Pushchino State Institute of Natural Sciences, Pushchino, Russian Federation.

e-mail: natasha.goryacheva2017@yandex.ru

Russian Federation

Dmitrij Ivanovic Rzhevskiy

Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences

Email: rjevski@bibch.ru
ORCID iD: 0000-0003-0520-4021

Кандидат биол. наук, старший научный сотрудник лаборатории биологических испытаний ФИБХ им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук, 142290, Пущино, Российская Федерация.

e-mail: rjevski@bibch.ru 

Russian Federation

Gulsara Amsngalievna Slashheva

Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences

Email: slashcheva_ga@mail.ru
ORCID iD: 0000-0002-4422-487X

Кандидат биол. наук, старший научный сотрудник лаборатории биологических испытаний, ФИБХ им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук, 142290, Пущино, Российская Федерация.

e-mail: slashcheva_ga@mail.ru 

Russian Federation

Nadezhda Ivanovna Novikova

Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences

Email: novikova@bibsh.ru
ORCID iD: 0000-0002-9145-2736

Кандидат биол. наук, старший научный сотрудник лаборатории биологических испытаний, ФИБХ им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук, 142290, Пущино, Российская Федерация.

e-mail: novikova@bibsh.ru 

Russian Federation

Mikhail Valentinivich Kisilevskiy

N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Healthcare of the Russian Federation

Email: kisele@inbox.ru
ORCID iD: 0000-0002-0132-167X

Доктор мед. наук, профессор, руководитель лаборатории клеточного иммунитета ФГБНУ «Национальный медицинский исследовательский центр онкологии имени Н.Н. Блохина» Минздрава России, 115478, Москва, Российская Федерация.

e-mail: kisele@inbox.ru 

Russian Federation

Irina Olegovna Chikileva

N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Healthcare of the Russian Federation

Email: irinatchikileva@mail.ru
ORCID iD: 0000-0003-0769-1695

Кандидат биол. наук, младший научный сотрудник лаборатории клеточного иммунитета ФГБУ «Национальный медицинский исследовательский центр онкологии имени Н.Н. Блохина» Минздрава России, 115478, Москва, Российская Федерация.

e-mail: irinatchikileva@mail.ru 

Russian Federation

Rajmonda Yanovna Vlasenko

N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Healthcare of the Russian Federation

Email: vlasenko2002@bk.ru
ORCID iD: 0000-0002-3560-001X

Кандидат биол. наук, старший научный сотрудник лаборатории клеточного иммунитета ФГБУ «Национальный медицинский исследовательский центр онкологии имени Н.Н. Блохина» Минздрава России, Москва, Российская Федерация.

e-mail: vlasenko2002@bk.ru 

Russian Federation

Igor Aleksandrovich Dyachenko

Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences; Pushchino State Institute of Natural Sciences

Email: dyachenko@bibch.ru
ORCID iD: 0000-0002-3053-2804

Кандидат биол. наук, заведующий лабораторией, старший научный сотрудник лаборатории биологических испытаний, ФИБХ им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук, доцент ФГБОУ ВО
Пущинского государственного естественно-научного института, 142290, Пущино, Российская Федерация.

e-mail: dyachenko@bibch.ru 

Russian Federation

Arkadij Nikolaevich Murashev

Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences; Pushchino State Institute of Natural Sciences

Email: murashev@bibch.ru
ORCID iD: 0000-0002-2279-9285

Доктор биол. наук, профессор., руководитель, ведущий научный сотрудник лаборатории биологических испытаний ФИБХ им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук, декан БиоМедФармТехнологического факультета ФГБОУ ВО Пущинского государственного естественно-научного института, 142290, Пущино, Российская Федерация.

e-mail: murashev@bibch.ru 

Russian Federation

Dmitrij Aleksandrovich Bondarenko

Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences

Email: bondarenko@bibch.ru
ORCID iD: 0000-0002-8055-0066

Кандидат биол. наук, старший научный сотрудник лаборатории биологических испытаний ФИБХ им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук, 142290, Пущино, Российская Федерация.

e-mail: bondarenko@bibch.ru 

Russian Federation

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Copyright (c) 2022 Goryacheva N.A., Rzhevskiy D.I., Slashheva G.A., Novikova N.I., Kisilevskiy M.V., Chikileva I.O., Vlasenko R.Y., Dyachenko I.A., Murashev A.N., Bondarenko D.A.


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