Direct comparison of KATUSHKA far-red fluorescent protein variants with luciferase bioluminescence in xenograft mouse models of epithelial tumor studies
K. A. Levchuk,
D. A. Bogdanova,
A. Ershova,
S. V. Smirnov,
A. A. Goldaeva,
M. L. Vasuytina,
M. S. Istomina,
O. N. Demidov,
A. V. Petukhov,
Ya. G. Toropova https://doi.org/10.18705/2311-4495-2022-9-6-59-70
Abstract
Background. Research in oncology assumes establishment and usage of xenograft animal models, meeting the requirements of humanity and rationality. Although praised as promising field of research, preclinical studies of low-molecular-weight inhibitors, antitumor monoclonal antibodies, vaccines, cellular therapy products, CAR-T cells entail continuous control of tumor growth dynamics. Luciferase detection of bioluminescence requires injection of a reporter substrate. However, injections not only are laborious, time-consuming and expensive, but are also stressful for animals. Thus, a vast variety of new visualization methods is employed, including proteins of the far-red spectrum.
Objective. The study aimed to compare detection efficiency of tumor growth dynamics in mice models of cervical cancer, applying a commercially available line of fluorescent red protein derivatives of Katushka clade and Renilla luciferase, that is commonly used in in vivo studies.
Design and methods. Xenograft mice models were derived injecting modified HeLa cell line, that expresses fluorescent reporter proteins: Katushka, Katushka2S, TurboRFP, TurboFP650 and Renilla enzyme.
Results. Spectral properties and emission wavelength of far-red fluorescent protein Katushka and Katushka2S outlines these markers within RFP derivatives lineage as outstanding instrument for in vivo tumor visualization.
Conclusion. Detection of fluorescent far-red reporters Katushka and Katushka2S can be considered as a credible alternative to Renilla luciferase bioluminescence in experimental models in vivo on the part of immunotherapy research.
Keywords
bioluminescence,
fluorescence in biomedical research,
fluorescent proteins,
luciferase detection,
visualization of xenografts in vivo
Supporting agencies: Recent work was funded by the Russian Science Foundation No. 19-74-20026 dated April 16, 2019 “Investigation of the effectiveness of T-cells with a chimeric antigen receptor (CAR-T) against cell and animal models of solid tumors” 2019-2022.
About the Authors
K. A. Levchuk
Almazov National Medical Research Centre
Ksenia A. Levchuk, PhD student, Junior researcher
Saint Petersburg
D. A. Bogdanova
Institute of Cytology, Russian Academy of Science; Sirius University of Science and Technology
Daria A. Bogdanova, Senior laboratory researcher; Junior researcher, PhD student
Saint Petersburg
Sochi
A. Ershova
Almazov National Medical Research Centre; Sirius University of Science and Technology
Daria A. Bogdanova, Senior laboratory researcher; Junior researcher, PhD student
Saint Petersburg
Sochi
S. V. Smirnov
Almazov National Medical Research Centre
Sergey V. Smirnov, Junior researcher
Saint Petersburg
A. A. Goldaeva
Institute of Experimental Medicine
Alexandra A. Goldaeva, student
Saint Petersburg
M. L. Vasuytina
Almazov National Medical Research Centre
Marina L. Vasyutina, Researcher, Vivarium veterinarian
Saint Petersburg
M. S. Istomina
Almazov National Medical Research Centre
Maria S. Istomina, Junior researcher
Saint Petersburg
O. N. Demidov
Almazov National Medical Research Centre; Institute of Cytology, Russian Academy of Science; Sirius University of Science and Technology
Oleg N. Demidov, MD, Leading researcher; Leading researcher, Sirius University of Science and Technology
Saint Petersburg
Sochi
A. V. Petukhov
Almazov National Medical Research Centre
Alexey V. Petukhov, researcher
Saint Petersburg
Ya. G. Toropova
Almazov National Medical Research Centre; Institute of Experimental Medicine
Yana G. Toropova, Ph.D., assistant professor, Department of pathological physiology, Institute of medical education, Head of the Bioprosthetics and Cardioprotection Research Laboratory; deputy director of scientific work
Saint Petersburg
References
1.
Review
For citations:
Levchuk K.A., Bogdanova D.A., Ershova A., Smirnov S.V., Goldaeva A.A., Vasuytina M.L., Istomina M.S., Demidov O.N., Petukhov A.V., Toropova Ya.G. Direct comparison of KATUSHKA far-red fluorescent protein variants with luciferase bioluminescence in xenograft mouse models of epithelial tumor studies. Translational Medicine. 2022;9(6):59-70. (In Russ.) https://doi.org/10.18705/2311-4495-2022-9-6-59-70
Views:
622
Email this article 