Gastric cancer modeling in immunodeficient mice with orthotopic xenotransplantation

Purpose of the study. Creation of a transplantable orthotopic PDX model of gastric cancer in Balb/c Nude immunodeficient mice using implantation and injection.

Materials and methods. Two methods, that are injection and implantation, were used to create an orthotopic PDX model of human gastric cancer. The first method involved injections of a suspension of a mechanically disaggregated patient's tumor after filtration into the gastric wall of Balb/c Nude mice. For the second method, small fragments (3 × 3 × 3 mm) of patients' tumors were implanted in the gastric wall of mice along the greater curvature with a dissection of the serous muscular layer.

Results. Control laparotomy in Balb/c Nude immunodeficient mice showed a successful engraftment of the tumor material at the 1st and 3rd procedures when using the implantation method for the creation of a PDX model of gastric cancer. The injection method was ineffective, and no models were created. The histological type of the obtained PDX models was compared to the type of the donor tumor by histological examination (hematoxylin and eosin staining). The tumor grade remained stable and did not change during xenograft passage, which showed that the obtained model was identical to the histotype of the donor tumor.

Conclusion. The presented implantation method for the model creation results in effective tumor engraftment. The developed model can be used to test the effectiveness of anticancer or antimetastatic drugs, for studying the functions of biomarkers, or in assessing the microenvironment of a gastric cancer.

1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021 May;71(3):209–249. https://doi.org/10.3322/caac.21660

2. Kit OI, Kasatkin VF, Maksimov AYu, Mamulyan HG. Combined gastrectomy at locally-advanced gastric cancer after neoadjuvant polychemotherapy. News of higher educational institutions. The North Caucasus region. Natural sciences. 2012;(4):108–111. (In Russ.). EDN: PCIYTL

3. Bernards N, Creemers GJ, Nieuwenhuijzen G a. P, Bosscha K, Pruijt JFM, Lemmens VEPP. No improvement in median survival for patients with metastatic gastric cancer despite increased use of chemotherapy. Ann Oncol. 2013 Dec;24(12):3056– 3060. https://doi.org/10.1093/annonc/mdt401

4. Corso S, Isella C, Bellomo SE, Apicella M, Durando S, Migliore C, et al. A Comprehensive PDX Gastric Cancer Collection Captures Cancer Cell-Intrinsic Transcriptional MSI Traits. Cancer Res. 2019 Nov 15;79(22):5884–5896. https://doi.org/10.1158/0008-5472.CAN-19-1166

5. Kit OI, Vashchenko LN, Dashkova IR, Kutilin DS, Maksimov AYU, Goncharova AS. Xenograft models of human breast cancer in experimental studies. Modern problems of science and education. 2019;(6):184–184. (In Russ.). EDN: OWQEZP

6. Hoffman RM. Orthotopic metastatic (MetaMouse) models for discovery and development of novel chemotherapy. Methods Mol Med. 2005;111:297–322. https://doi.org/10.1385/1-59259-889-7:297

7. Julien S, Merino-Trigo A, Lacroix L, Pocard M, Goéré D, Mariani P, et al. Characterization of a large panel of patient-derived tumor xenografts representing the clinical heterogeneity of human colorectal cancer. Clin Cancer Res. 2012 Oct 1;18(19):5314–5328. https://doi.org/10.1158/1078-0432.CCR-12-0372

8. Kiblitskaya AA, Karasev TS, Goncharova AS, Maksimov AYu. Methods for modeling tumor growth in mice in experimental studies of human gastric cancer. South Russian Journal of Cancer. 2021;2(4):26–37. (In Russ.). https://doi.org/10.37748/2686-9039-2021-2-4-4, EDN: BKTBYG

9. Busuttil RA, Liu DS, Di Costanzo N, Schröder J, Mitchell C, Boussioutas A. An orthotopic mouse model of gastric cancer invasion and metastasis. Sci Rep. 2018 Jan 16;8(1):825. https://doi.org/10.1038/s41598-017-19025-y

10. Reddavid R, Corso S, Moya-Rull D, Giordano S, Degiuli M. Patient-Derived Orthotopic Xenograft models in gastric cancer: a systematic review. Updates Surg. 2020 Dec;72(4):951–966. https://doi.org/10.1007/s13304-020-00751-4

11. Guidelines for the maintenance and use of laboratory animals. Ed. by I. V. Belozertseva, D. V. Blinova, M. S. Krasilshchikova. Eighth edition. Moscow: Limited Liability Company "IRBIS", 2017, 304 p. (In Russ.). EDN: ZRJVDJ

12. Kuwata T, Yanagihara K, Iino Y, Komatsu T, Ochiai A, Sekine S, et al. Establishment of Novel Gastric Cancer Patient-Derived Xenografts and Cell Lines: Pathological Comparison between Primary Tumor, Patient-Derived, and Cell-Line Derived Xenografts. Cells. 2019 Jun 14;8(6):585. https://doi.org/10.3390/cells8060585

13. Kang W, Maher L, Michaud M, Bae SW, Kim S, Lee HS, et al. Development of a Novel Orthotopic Gastric Cancer Mouse Model. Biol Proced Online. 2021 Jan 4;23(1):1. https://doi.org/10.1186/s12575-020-00137-1