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Disruption of the Tff1 gene in mice using CRISPR/Cas9 promotes body weight reduction and gastric tumorigenesis

Abstract

Trefoil factor 1 (TFF1, also known as pS2) is strongly expressed in the gastrointestinal mucosa and plays a critical role in the differentiation of gastric glands. Since approximately 50% of all human gastric cancers are associated with decreased TFF1 expression, it is considered a tumor suppressor gene. Tff1 deficiency in mice results in histological changes in the antral and pyloric gastric mucosa, with severe hyperplasia and dysplasia of epithelial cells, resulting in the development of antropyloric adenoma. Here, we generated Tff1-knockout (KO) mice, without a neomycin resistant (NeoR) cassette, using the clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRSIPR/Cas9) system. Though our Tff1-KO mice showed phenotypes very similar to the previous embryonic stem (ES)-cell-based KO mice, they differed from the previous reports in that a reduction in body weight was observed in males. These results demonstrate that these newly established Tff1-KO mice are useful tools for investigating genetic and environmental factors influencing gastric cancer, without the effects of artificial gene insertion. Furthermore, these findings suggest a novel hypothesis that Tff1 expression influences gender differences.

References

  1. 1.

    Roder DM. The epidemiology of gastric cancer. Gastric Cancer 2002; 5 Suppl 1: 5–11.

  2. 2.

    Kamangar F, Dores GM, Anderson WF. Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol 2006; 24(14): 2137–2150.

  3. 3.

    Muoz N, Franceschi S. Epidemiology of gastric cancer and perspectives for prevention. Salud Publica Mex 1997; 39(4): 318–330.

  4. 4.

    Hohenberger P, Gretschel S. Gastric cancer. Lancet 2003; 362(9380): 305–315.

  5. 5.

    Shi SQ, Cai JT, Yang JM. Expression of trefoil factors 1 and 2 in precancerous condition and gastric cancer. World J Gastroenterol 2006; 12(19): 3119–3122.

  6. 6.

    Jakowlew SB, Breathnach R, Jeltsch JM, Masiakowski P, Chambon P. Sequence of the pS2 mRNA induced by estrogen in the human breast cancer cell line MCF-7. Nucleic Acids Res 1984; 12(6): 2861–2878.

  7. 7.

    Xiao P, Ling H, Lan G, Liu J, Hu H, Yang R. Trefoil factors: Gastrointestinal-specific proteins associated with gastric cancer. Clin Chim Acta 2015; 450: 127–134.

  8. 8.

    Lefebvre O, Chenard MP, Masson R, Linares J, Dierich A, LeMeur M, Wendling C, Tomasetto C, Chambon P, Rio MC. Gastric mucosa abnormalities and tumorigenesis in mice lacking the pS2 trefoil protein. Science 1996; 274(5285): 259–262.

  9. 9.

    Ribieras S, Tomasetto C, Rio MC. The pS2/TFF1 trefoil factor, from basic research to clinical applications. Biochim Biophys Acta 1998; 1378(1): F61–77.

  10. 10.

    Rio MC, Bellocq JP, Daniel JY, Tomasetto C, Lathe R, Chenard MP, Batzenschlager A, Chambon P. Breast cancer-associated pS2 protein: synthesis and secretion by normal stomach mucosa. Science 1988; 241(4866): 705–708.

  11. 11.

    Wright AV, Nuez JK, Doudna JA. Biology and Applications of CRISPR Systems: Harnessing Nature’s Toolbox for Genome Engineering. Cell 2016; 164(1-2): 29–44.

  12. 12.

    Lee JH, Park JH, Nam TW, Seo SM, Kim J Y, Lee HK, Han JH, Park S Y, Choi YK, Lee HW. Differences between immunodeficient mice generated by classical gene targeting and CRISPR/Cas9-mediated gene knockout. Transgenic Res 2018; 27(3): 241–251.

  13. 13.

    Roh JI, Lee J, Park SU, Kang YS, Lee J, Oh AR, Choi DJ, Cha J Y, Lee HW. CRISPR-Cas9-mediated generation of obese and diabetic mouse models. Exp Anim 2018; 67(2): 229–237.

  14. 14.

    Sung YH, Kim JM, Kim HT, Lee J, Jeon J, Jin Y, Choi JH, Ban YH, Ha SJ, Kim CH, Lee HW, Kim JS. Highly efficient gene knockout in mice and zebrafish with RNA-guided endonucleases. Genome Res 2014; 24(1): 125–131.

  15. 15.

    Henry JA, Bennett MK, Piggott NH, Levett DL, May FE, Westley BR. Expression of the pNR-2/pS2 protein in diverse human epithelial tumours. Br J Cancer 1991; 64(4): 677–682.

  16. 16.

    Luqmani Y, Bennett C, Paterson I, Corbishley CM, Rio MC, Chambon P, Ryall G. Expression of the pS2 gene in normal, benign and neoplastic human stomach. Int J Cancer 1989; 44(5): 806–812.

  17. 17.

    Machado JC, Carneiro F, Ribeiro P, Blin N, Sobrinho-Simes M. pS2 protein expression in gastric carcinoma. An immunohistochemical and immunoradiometric study. Eur J Cancer 1996; 32A(9): 1585–1590.

  18. 18.

    Theisinger B, Welter C, Seitz G, Rio MC, Lathe R, Chambon P, Blin N. Expression of the breast cancer associated gene pS2 and the pancreatic spasmolytic polypeptide gene (hSP) in diffuse type of stomach carcinoma. Eur J Cancer 1991; 27(6): 770–773.

  19. 19.

    Wanebo HJ, Kennedy BJ, Chmiel J, Steele G Jr, Winchester D, Osteen R. Cancer of the stomach. A patient care study by the American College of Surgeons. Ann Surg 1993; 218(5): 583–592.

  20. 20.

    Everett SM, Axon AT. Early gastric cancer in Europe. Gut 1997; 41(2): 142–150.

  21. 21.

    Saukkonen K, Tomasetto C, Narko K, Rio MC, Ristimki A. Cyclooxygenase-2 expression and effect of celecoxib in gastric adenomas of trefoil factor 1-deficient mice. Cancer Res 2003; 63(12): 3032–3036.

  22. 22.

    Blum D, Stene GB, Solheim TS, Fayers P, Hjermstad MJ, Baracos VE, Fearon K, Strasser F, Kaasa S. Euro-Impact. Validation of the Consensus-Definition for Cancer Cachexia and evaluation of a classification model—a study based on data from an international multicentre project (EPCRC-CSA). Ann Oncol 2014; 25(8): 1635–1642.

  23. 23.

    Artelt P, Grannemann R, Stocking C, Friel J, Bartsch J, Hauser H. The prokaryotic neomycin-resistance-encoding gene acts as a transcriptional silencer in eukaryotic cells. Gene 1991; 99(2): 249–254.

  24. 24.

    Valera A, Perales JC, Hatzoglou M, Bosch F. Expression of the neomycin-resistance (neo) gene induces alterations in gene expression and metabolism. Hum Gene Ther 1994; 5(4): 449–456.

  25. 25.

    Alitalo K, Koskinen P, Mkel TP, Saksela K, Sistonen L, Winqvist R. myc oncogenes: activation and amplification. Biochim Biophys Acta 1987; 907(1): 1–32.

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Correspondence to Jaehoon Lee or Ki Taek Nam or Han-Woong Lee.

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This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://doi.org/creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Kim, H., Jeong, H., Cho, Y. et al. Disruption of the Tff1 gene in mice using CRISPR/Cas9 promotes body weight reduction and gastric tumorigenesis. Lab Anim Res 34, 257–263 (2018). https://doi.org/10.5625/lar.2018.34.4.257

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Keywords

  • Tff1
  • gastric cancer
  • CRISPR/Cas9
  • Tff1-knockout mouse