Human Erythropoietin Induces Lung Failure and Erythrocytosis in Transgenic Mice

  • Kim, Myoung Ok (School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Kim, Sung Hyun (School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Shin, Mi Jung (School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Lee, Dong Beom (The Republic of Korea Red Cross, Nam-Bu Seoul Blood Center) ;
  • Kim, Tae Won (Department of Life Science, Dongguk University) ;
  • Kim, Kil Soo (Department of Veterinary Medicine, College of Veterinary Medicine, Kyungpook National University) ;
  • Ha, Ji Hong (School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Lee, Sanggyu (School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Park, Yong Bok (School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Kim, Sun Jung (Department of Life Science, Dongguk University) ;
  • Ryoo, Zae Young (School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University)
  • Received : 2006.09.06
  • Accepted : 2006.11.16
  • Published : 2007.02.28

Abstract

We have expressed human erythropoietin (EPO) in transgenic mice using a recombinant EPO cDNA combined with a partial TPO construct. The gene was microinjected using standard techniques and five mice were detected as transgenic by PCR and further used as founders. The life span of the transgenic founders was much shorter than that of their normal littermates. Most of the tissues of the transgenic founders contained human EPO transcripts as judged by RT-PCR. Especially high expression levels were seen in the liver and lung. EPO protein levels in serum were examined by ELISA and ranged from 266-414 mIU/ml. The number of red blood cell, white blood cell and hemoglobin in the hEPO transgenic mice was higher than in normal mice. These results indicate that overexpression of hEPO is deleterious and can provoke lung failure and erythrocytosis.

Keywords

Acknowledgement

Supported by : The Korea Ministry of Agriculture and Fisheries, Korea Science and Engineering Foundation

References

  1. Aguirre, A., Castro-Palomino, N., De la Fuente, J., and Ovidio Castro, F. O. (1998) Expression of human erythropoietin transgenes and of the endogenous WAP gene in the mammary gland of transgenic rabbits during gestation and lactation. Trans. Res. 7, 311–317
  2. Alexander, D., Hunpei, S., Dimitri, S., Wilfried, B., Klaus, F. W., et al. (2004) Norepinephrine-induced acute heart failure in transgenic mice overexpressing erythropoietin. Cardi. Res. 61, 105−114
  3. Archibald, A. L., McClenaghan, M., Hornsey, V., Simons, J. P., and Clark, A. L. (1990) High-level expression of biologically active human ${\alpha}1$-antitrypson in the milk of transgenic mice. Proc. Natl. Acad. Sci. USA 87, 5178−5182
  4. Barash, I., Faerman, A., Ratovitsky, T., Puzis, R., Nathan, M., et al. (1994) Ectopic expression of $\beta$-lactoglobulin/human serum albumin fusion genes in transgenic mice: hormonal regulation and in situ localization. Trans. Res. 3, 141−151
  5. Faulds, D. and Sorkin, E. M. (1989) Epoetin (recombinant human erythropoietin). Drugs 38, 863−899
  6. Follis, A. G. and Netsky, M. G. (1969) Astrocytes in formalinfixed paraffin sections. A new modification of Cajal's technic and comparison with the hematoxylin and eosin method. Tech. Bull. Regist. Med. Technol. 39, 38−41
  7. Fried, W. (1972) The liver as a source of extra renal erythropoietin production. Blood 40, 671
  8. Halina, M. Z., Nadia, S., Reza, B., Rosemary, C., Roman, D., et al. (2002) The use of the uromodulin promoter to target production of recombinant proteins into urine of transgenic animals. Trans. Res. 11, 425−435
  9. Jacobs, K., Shoemarker, C., Rudersdorf, R., Neil, S. D., and Kaufman, R. J. (1985) Isolation and characterization of genomic and cDNA clones of human erythropoietin. Nature 313, 806−810 https://doi.org/10.1038/313810a0
  10. Jacobson, L. O., Goldwasswe, E., Fried, W., and Plazak, L. (1957) Role of the kidney in erythropoiesis. Nature 179, 633−634 https://doi.org/10.1038/179634a0
  11. Jelkmann, W. (1992) Erythropoietin: structure, control of production, and function. Physiol. Rev. 72, 449−489
  12. Kantz, S. B. (1991) Erythropoietin. Blood 77, 419−434
  13. Kim, T. W., Ji, J. W., Chang, H. G., Kim, M. O., Ryoo, Z. Y., et al. (2005) 3′-half of the thrombopoietin cDNA confers higher expression pf erythropoietin at the RNA but not at the protein level. Mol. Cells 19, 198−204
  14. Korhonen, V. P., Tolvanen, M., Hyttinen, J. M., Uusi-Oukari, M., Sinervirta, R., et al. (1997) Expression of bovine betalactoglobulin/ human erythropoietin fusion protein in the milk of transgenic mice and rabbits. Eur. J. Biochem. 245, 482− 489
  15. Koury, M. J., Bondurant, M. C., and Koury, M. J. (1990) Erythropoietin retards DNA breakdown and prevents programmed cell death in erythroid progenitor cells. Science 248, 378−381 https://doi.org/10.1126/science.248.4953.381
  16. Lee, S. S., Moon, J. H., Ha, J. W., Lee, Y. K., Ahn, J. S., et al. (2004) A case of transition of polycythemia vera to chronic neutrophilic leukemia. Korean J. Intern. Med. 19, 285−288
  17. Madan, A., Lin, C., Hatch, II S. L., and Curtin, P. T. (1995) Regulated basal, inducible, and tissue-specific human erythropoietin gene expression in transgenic mice requires multiple cis DNA sequences. Blood 85, 2735−2741
  18. Massoud, M., Attal, J., Thepot, D., Pointu, H., Stinnakre, M. G., et al. (1996) The deleterious effects of human erythropoietin gene driven by the rabbit whey acidic protein gene promoter in transgenic rabbits. Reprod. Nutr. Dev. 36, 555–563
  19. Miyamoto, Y., Kuramitsu-Miyamoto, K., Iwanaga, E., Uchio- Yamada, K., Yamaguchi-Yamada, M., et al. (2005) Effect of human erythropoietin (hEPO) treatment on anemia in ICRderived glomerulonephritis (ICGN) mice. Exp. Anim. 54, 181−184
  20. Richard, P., Pollard, H., Lanctin, C., Bello-Roufai, M., Desigaux, L., et al. (2005) Inducible production of erythropoietin using intramuscular injection of block copolymer/DNA formulation. J. Gene Med. 7, 80−86
  21. Sawyer, S. T., Krantz, S. B., and Goldwasser, E. (1987) Binding and receptor mediated endocytosis of erythropoietin in friend virus infected erythoid cells. J. Biol. Chem. 262, 5554−5562
  22. Semenza, G. L., Traystman, M. D., Gearhart, J. D., and Antonarakis, S. E. (1989) Polycythemia in transgenic mice expressing the human erythropoietin gene. Proc. Natl. Acad. Sci. USA 86, 2301−2305
  23. Sohn, B. H., Kim, S. J., Park, H., Park, S. K., and Lee, S. C. (1999) Expression and characterization of bioactive human thrombopoietin in the milk of transgenic mice. DNA Cell Biol. 18, 845−852
  24. Suk, K., Jung, D. Y., Kang, S. K., Kang, S. W., Seo, E. J., et al. (1995) Human erythropoietin-induced polycysthemia in transgenic mice. Mol. Cells 5, 634–640
  25. Vogel, J., Kiessling, I., Heinicke, K., Stallmach, T., Ossent, P., et al. (2003) Transgenic mice overexpressing erythropoietin adapt to excessive erythrocytosis by regulating blood viscosity. Blood 15, 2278−2284
  26. Wanger, K. F., Katschinski, D. M., Hasegawa, J., Schumacher, D., Meller, B., et al. (2001) Chronic inborn erythrocytosis leads to cardiac dysfunction and premature death in mice overexpressing erythropoietin. Blood 15, 536−542
  27. Yamaguchi, K., Jurgens, K. D., Bartels, H., Scheid, P., and Piier, J. (1988) Dependence of O2 transfer conductance of red blood cells on cellular dimensions. Adv. Exp. Med. Biol. 222, 571−578
  28. Zanjani, E. D., Poster, J., Burlington, H., Mann, L. I., and Wasserman, L. R. (1977) Liver as the primary site for erythropoietin formation in the fetus. J. Lab. Clin. Med. 89, 640−644