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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Enhanced biological effects of Phe140Asn, a novel human granulocyte colony-stimulating factor mutant, on HL60 cells

  • Chung, Hee-Kyoung (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kim, Sung-Woo (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Byun, Sung-June (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Ko, Eun-Mi (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Chung, Hak-Jae (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Woo, Jae-Seok (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Yoo, Jae-Gyu (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Lee, Hwi-Cheul (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Yang, Byoung-Chul (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kwon, Moo-Sik (Department of Genetic Engineering, Sungkyunkwan University) ;
  • Park, Soo-Bong (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Park, Jin-Ki (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kim, Kyung-Woon (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration)
  • Received : 2011.06.08
  • Accepted : 2011.06.23
  • Published : 2011.10.31
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Abstract

Granulocyte colony-stimulating factor (G-CSF) is a cytokine secreted by stromal cells and plays a role in the differentiation of bone marrow stem cells and proliferation of neutrophils. Therefore, G-CSF is widely used to reduce the risk of serious infection in immunocompromised patients; however, its use in such patients is limited because of its non-persistent biological activity. We created an N-linked glycosylated form of this cytokine, hG-CSF (Phe140Asn), to assess its biological activity in the promyelocyte cell line HL60. Enhanced biological effects were identified by analyzing the JAK2/STAT3/survivin pathway in HL60 cells. In addition, mutant hG-CSF (Phe140Asn) was observed to have enhanced chemoattractant effects and improved differentiation efficiency in HL60 cells. These results suggest that the addition of N-linked glycosylation was successful in improving the biological activity of hG-CSF. Furthermore, the mutated product appears to be a feasible therapy for patients with neutropenia.

Keywords

References

  1. Bishop, B., Koay, D. C., Sartorelli, A. C. and Regan, L. (2001) Reengineering granulocyte colony-stimulating factor for enhanced stability. J. Biol. Chem. 276, 33465-33470. https://doi.org/10.1074/jbc.M104494200
  2. Ribeiro, D., Veldwijk, M. R., Benner, A., Laufs, S., Wenz, F., Ho, A. D. and Fruehauf, S. (2007) Differences in functional activity and antigen expression of granulocytes primed in vivo with filgrastim, lenograstim, or pegfilgrastim. Transfusion 47, 969-980. https://doi.org/10.1111/j.1537-2995.2007.01241.x
  3. Burgess, A. W. and Metcalf, D. (1980) The nature and action of granulocyte-macrophage colony stimulating factors. Blood 56, 947-958.
  4. Hill, C. P., Osslund, T. D. and Eisenberg, D. (1993) The structure of granulocyte-colony-stimulating factor and its relationship to other growth factors. Proc. Natl. Acad. Sci. U.S.A. 90, 5167-5171. https://doi.org/10.1073/pnas.90.11.5167
  5. Tamada, T., Honjo, E., Maeda, Y., Okamoto, T., Ishibashi, M., Tokunaga, M. and Kuroki, R. (2006) Homodimeric cross-over structure of the human granulocyte colony-stimulating factor (GCSF) receptor signaling complex. Proc. Natl. Acad. Sci. U.S.A. 103, 3135-3140. https://doi.org/10.1073/pnas.0511264103
  6. Jiang, K., Hein, N., Eckert, K., Luscher-Firzlaff, J. and Luscher, B. (2008) Regulation of the MAD1 promoter by G-CSF. Nucleic Acids Res. 36, 1517-1531. https://doi.org/10.1093/nar/gkn002
  7. Gu, L., Chiang, K. Y., Zhu, N., Findley, H. W. and Zhou, M. (2007) Contribution of STAT3 to the activation of survivin by GM-CSF in CD34+ cell lines. Exp. Hematol. 35, 957-966. https://doi.org/10.1016/j.exphem.2007.03.007
  8. Chakraborty, A., White, S. M., Schaefer, T. S., Ball, E. D., Dyer, K. F. and Tweardy, D. J. (1996) Granulocyte colony-stimulating factor activation of Stat3 alpha and Stat3 beta in immature normal and leukemic human myeloid cells. Blood 88, 2442-2449.
  9. Yin, S., Wang, R., Zhou, F., Zhang, H. and Jing, Y. (2011) Bcl-XL is a dominant antiapoptotic protein which inhibits homoharringtonine-induced apoptosis in leukemia cells. Mol. Pharmacol. 79, 1072-1083 https://doi.org/10.1124/mol.110.068528
  10. Gervais, V., Zerial, A. and Oschkinat, H. (1997) NMR investigations of the role of the sugar moiety in glycosylated recombinant human granulocyte-colony-stimulating factor. Eur. J. Biochem. 247, 386-395. https://doi.org/10.1111/j.1432-1033.1997.00386.x
  11. Cumming, D. A. (1991) Glycosylation of recombinant protein therapeutics: control and functional implications. Glycobiology 1, 115-130. https://doi.org/10.1093/glycob/1.2.115
  12. Lu, H. S., Boone, T. C., Souza, L. M. and Lai, P. H. (1989) Disulfide and secondary structures of recombinant human granulocyte colony stimulating factor. Arch. Biochem. Biophys. 268, 81-92. https://doi.org/10.1016/0003-9861(89)90567-5
  13. Ishikawa, M., Iijima, H., Satake-Ishikawa, R., Tsumura, H., Iwamatsu, A., Kadoya, T., Shimada, Y., Fukamachi, H., Kobayashi, K., Matsuki, S. and Asano, K. (1992) The substitution of cysteine 17 of recombinant human G-CSF with alanine greatly enhanced its stability. Cell Struct. Funct. 17, 61-65. https://doi.org/10.1247/csf.17.61
  14. Shimozaki, K., Nakajima, K., Hirano, T. and Nagata, S. (1997) Involvement of STAT3 in the granulocyte colony-stimulating factor-induced differentiation of myeloid cells. J. Biol. Chem. 272, 25184-25189. https://doi.org/10.1074/jbc.272.40.25184
  15. Zaker, F., Oody, A. and Arjmand, A. (2007) A study on the antitumoral and differentiation effects of peganum harmala derivatives in combination with ATRA on leukaemic cells. Arch. Pharm. Res. 30, 844-849. https://doi.org/10.1007/BF02978835
  16. Detmers, P. A., Thieblemont, N., Vasselon, T., Pironkova, R., Miller, D. S. and Wright, S. D. (1996) Potential role of membrane internalization and vesicle fusion in adhesion of neutrophils in response to lipopolysaccharide and TNF. J. Immunol. 157, 5589-5596.
  17. Avalos, B. R. (1996) Molecular analysis of the granulocyte colony-stimulating factor receptor. Blood 88, 761-777.
  18. Gascan, H., Moreau, J. F., Jacques, Y. and Soulillou, J. P. (1989) Response of murine IL3-sensitive cell lines to cytokines of human and murine origin. Lymphokine Res. 8, 79-84.
  19. Shimoda, K., Iwasaki, H., Okamura, S., Ohno, Y., Kubota, A., Arima, F., Otsuka, T. and Niho, Y. (1994) G-CSF induces tyrosine phosphorylation of the JAK2 protein in the human myeloid G-CSF responsive and proliferative cells, but not in mature neutrophils. Biochem. Biophys. Res. Commun. 203, 922-928. https://doi.org/10.1006/bbrc.1994.2270
  20. Li, J. and Sartorelli, A. C. (1994) Evidence for the glycosylation of the granulocyte colony-stimulating factor receptor. Biochem. Biophys. Res. Commun. 205, 238-244. https://doi.org/10.1006/bbrc.1994.2655
  21. Dangoor, D., Biondi, B., Gobbo, M., Vachutinski, Y., Fridkin, M., Gozes, I. and Rocchi, R. (2008) Novel glycosylated VIP analogs: synthesis, biological activity, and metabolic stability. J. Pept. Sci. 14, 321-328. https://doi.org/10.1002/psc.932
  22. Orlic, D., Kajstura, J., Chimenti, S., Limana, F., Jakoniuk, I., Quaini, F., Nadal-Ginard, B., Bodine, D. M., Leri, A. and Anversa, P. (2001) Mobilized bone marrow cells repair the infarcted heart, improving function and survival. Proc. Natl. Acad. Sci. U.S.A. 98, 10344-10349. https://doi.org/10.1073/pnas.181177898
  23. Gomez-Cambronero, J., Horn, J., Paul, C. C. and Baumann, M. A. (2003) Granulocyte-macrophage colony-stimulating factor is a chemoattractant cytokine for human neutrophils: involvement of the ribosomal p70 S6 kinase signaling pathway. J. Immunol. 171, 6846-6855. https://doi.org/10.4049/jimmunol.171.12.6846
  24. Kojima, H., Otani, A., Oishi, A., Makiyama, Y., Nakagawa, S. and Yoshimura, N. (2011) Granulocyte colony-stimulating factor attenuates oxidative stress-induced apoptosis in vascular endothelial cells and exhibits functional and morphologic protective effect in oxygen-induced retinopathy. Blood 117, 1091-1100. https://doi.org/10.1182/blood-2010-05-286963
  25. Pene, F., Claessens, Y. E., Muller, O., Viguie, F., Mayeux, P., Dreyfus, F., Lacombe, C. and Bouscary, D. (2002) Role of the phosphatidylinositol 3-kinase/Akt and mTOR/P70S6-kinase pathways in the proliferation and apoptosis in multiple myeloma. Oncogene 21, 6587- 6597. https://doi.org/10.1038/sj.onc.1205923

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