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

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

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