6 . 2023

Development of experimental mouse xenograft models of human tumors for preclinical in vivo studies of product for cellular immunotherapy

Abstract

Introduction. In connection with the active development of new directions in cellular immunotherapy, the question of approaches for conducting preclinical studies on safety and efficacy is acute. The most optimal for solving this problem is the use of tumor xenografts to simulate tumor growth in the body while maintaining homeostasis, metabolism and possible mechanisms of tolerance to a therapeutic agent. The effectiveness of this approach to the analysis of the effectiveness of cell preparations led to the need to standardize methodological approaches to the selection of human tumor cell lines and lines of immunodeficient mice.

The purpose of the study was to evaluate the effectiveness of xenograft formation in two strains of immunodeficient mice in order to obtain optimal models for preclinical studies of preparations of modified human cells.

Material and methods. To test the models we used mice of the NRG (NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ) and SCID lines and human tumor cell lines SW620 (human colon adenocarcinoma), U-87MG (human epithelial brain tumor), SK-MEL-37 (melanoma), NW-MEL-38 (melanoma) and S6 (melanoma). Experimental animals were injected subcutaneously into the region of the right shoulder blade with 100 µl of tumor cell suspension at doses of 1, 3 and 5 mln/mouse, followed by measurement of the size of the tumor node.

Results. The effectiveness of the formation of xenografts of various human cell lines in mice with immunodeficiency was evaluated. It has been shown that the NRG mice, compared to the SCID mice, has a higher ability to engraft tumor cell lines. And the cell lines themselves differ in the levels of expression of tumor-associated proteins and form a xenograft with different efficiency. The most optimal was the use of NRG mice and the SK-MEL-37 cell line, with a high level of expression of the GD2, NY-ESO-1 and MAGE-A4 antigens. An experiment on the treatment of solid tumors obtained after transplantation of human tumor cell lines into animals using anti-GD2-CAR-T cells showed effectiveness in reducing the size of the tumor node with intratumoral administration of modified human cells.

Conclusion. The results obtained confirm the efficiency of using xenografts in evaluating the effectiveness of the antitumor immune response of modified cell preparations in vivo.

Keywords:tumor; tumor xenografts; tumor cell lines; lines of immunodeficient mice; genetically modified T-lymphocytes

For citation: Lopatnikova J.A., Shevchenko J.A., Filippova J.G., Fisher M.S., Obleuhova I.A., Zavjalov E.L., Solovjeva O.I., Razumov I.A., Akahori Ya., Shiku H., Sennikov S.V. Development of experimental mouse xenograft models of human tumors for preclinical in vivo studies of product for cellular immunotherapy. Immunologiya. 2023; 44 (6): 709–20. DOI: https://doi.org/10.33029/0206-4952-2023-44-6-709-720 (in Russian)

Funding. The present study was supported by the Russian Science Foundation grant no. 21-65-00004, URL: https://rscf.ru/project/21-65-00004/

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

Authors’ contribution. The concept and design of the study – Lopatnikova Ju.A., Sennikov S.V.; work with cultures of cell lines – Shevchenko Ju.A., Filippova Ju.G., Fisher M.S.; work with primary human cells – Fisher M.S., Obleukhova I.A.; work with experimental animals – Zavyalov E.L., Solovieva O.I.; obtaining retroviral constructs – Akahori Ya., Shiku H.; statistical data processing – Lopatnikova Ju.A., Shevchenko Ju.A.; writing the text – Lopatnikova Ju.A., Shevchenko Ju.A.; text editing – Filippova Ju.G., Obleukhova I.A., Zavyalov E.L., approval of the final version of the article – Sennikov S.V.; responsibility for the integrity of all parts of the article – Lopatnikova Ju.A.

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