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Systemic overexpression of antizyme 1 in mouse reduces ornithine decarboxylase activity without major changes in tissue polyamine homeostasis

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Abstract

Polyamines, spermidine, spermine and their precursor putrescine, are ubiquitous cell components essential for normal cell growth. Increased polyamine levels and enhanced biosynthesis have been associated with malignant transformation and tumor formation, and thus, the polyamines have been considered to be a meaningful target to cancer therapies. However, clinical cancer treatment trials using inhibitors of polyamine synthesis have been unsuccessful probably due to compensatory uptake of polyamines from extracellular sources. The antizyme proteins regulate both polyamine biosynthesis and transport, and thus, the antizymes could provide an efficient approach to control cellular proliferation compared to the mere inhibition of biosynthesis. To define the role of antizymes in proliferative processes associated with the whole animal, we have generated transgenic mice overexpressing mouse antizyme 1 gene under its own regulatory sequences. Antizyme 1 protein was abundantly expressed in various organs and the expressed antizyme protein was functional as ornithine decarboxylase activity was significantly reduced in all tissues analyzed. However, antizyme 1 overexpression caused only minor changes in tissue polyamine levels demonstrating the challenges in using the “antizyme approach” to deplete polyamines in a living animal. Neither were there any changes in cellular proliferation in the proliferative tissues of transgenic animals. Interestingly though, there was occurrence of abnormally high level of apoptosis in the non-proliferating part of the colon epithelia. Otherwise, the transgenic founder mice appeared healthy and out of seven founders six were fertile. However, none of the founders could transmit the transgene suggesting that the antizyme 1 overexpression may be deleterious to transgenic gametes.

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References

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (2007) Current protocols in molecular biology. Wiley, Hoboken

    Google Scholar 

  • Bailey HH, Kim K, Verma AK et al (2010) A randomized, double-blind, placebo-controlled phase 3 skin cancer prevention study of α-difluoromethylornithine in subjects with previous history of skin cancer. Cancer Prev Res 3:35–47

    Article  CAS  Google Scholar 

  • Casero R Jr, Frydman B, Stewart TM, Woster PM (2005) Significance of targeting polyamine metabolism as an antineoplastic strategy: unique targets for polyamine analogues. Proc West Pharmacol Soc 48:24–30

    CAS  PubMed  Google Scholar 

  • Feith DJ, Shantz LM, Pegg AE (2001) Targeted antizyme expression in the skin of transgenic mice reduces tumor promoter induction of ornithine decarboxylase and decreases sensitivity to chemical carcinogenesis. Cancer Res 61:6073–6081

    CAS  PubMed  Google Scholar 

  • Fong LYY, Feith DJ, Pegg AE (2003) Antizyme overexpression in transgenic mice reduces cell proliferation, increases apoptosis, and reduces N-nitrosomethylbenzylamine-induced forestomach carcinogenesis. Cancer Res 63:3945–3954

    CAS  PubMed  Google Scholar 

  • Gritli-Linde A, Nilsson J, Bohlooly-Y M, Heby O, Linde A (2001) Nuclear translocation of antizyme and expression of ornithine decarboxylase and antizyme are developmentally regulated. Dev Dyn 220:259–275

    Article  CAS  PubMed  Google Scholar 

  • Gupta S, Ahmad N, Marengo SR, MacLennan GT, Greenberg NM, Mukhtar H (2000) Chemoprevention of prostate carcinogenesis by α-difluoromethylornithine in TRAMP mice. Cancer Res 60:5125–5133

    CAS  PubMed  Google Scholar 

  • Halmekytö M, Hyttinen JM, Sinervirta R et al (1991) Transgenic mice aberrantly expressing human ornithine decarboxylase gene. J Biol Chem 266:19746–19751

    PubMed  Google Scholar 

  • Heby O, Persson L, Rentala M (2007) Targeting the polyamine biosynthetic enzymes: a promising approach to therapy of African sleeping sickness, Chagas’ disease, and leishmaniasis. Amino Acids 33:359–366

    Article  CAS  PubMed  Google Scholar 

  • Hogan B, Constantini F, Lacy E (1986) Manipulating the mouse embryo. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 1–322

    Google Scholar 

  • Hyvönen T, Keinänen TA, Khomutov AR, Khomutov RM, Eloranta TO (1992) Monitoring of the uptake and metabolism of aminooxy analogues of polyamines in cultured cells by high-performance liquid chromatography. J Chromatogr 574:17–21

    Article  PubMed  Google Scholar 

  • Ivanov IP, Rohrwasser A, Terreros DA, Gesteland RF, Atkins JF (2000) Discovery of a spermatogenesis stage-specific ornithine decarboxylase antizyme: antizyme 3. Proc Natl Acad Sci USA 97:4808–4813

    Article  CAS  PubMed  Google Scholar 

  • Jänne J, Williams-Ashman HG (1971) On the purification of l-ornithine decarboxylase from rat prostate and effects of thiol compounds on the enzyme. J Biol Chem 246:1725–1732

    PubMed  Google Scholar 

  • Kahana C (2007) Ubiquitin dependent and independent protein degradation in the regulation of cellular polyamines. Amino Acids 33:225–230

    Article  CAS  PubMed  Google Scholar 

  • Kahana C, Asher G, Shaul Y (2005) Mechanisms of protein degradation: an odyssey with ODC. Cell Cycle 4:1461–1464

    Article  CAS  PubMed  Google Scholar 

  • Kaipia A, Toppari J, Mali P, Kangasniemi M, Alcivar AA, Hecht NB, Parvinen M (1990) Stage- and cell-specific expression of the ornithine decarboxylase gene during rat and mouse spermatogenesis. Mol Cell Endocrinol 73:45–52

    Article  CAS  PubMed  Google Scholar 

  • Kaiser A, Gottwald A, Maier W, Seitz HM (2003) Targeting enzymes involved in spermidine metabolism of parasitic protozoa—a possible new strategy for anti-parasitic treatment. Parasitol Res 91:508–516

    Article  CAS  PubMed  Google Scholar 

  • Kankare K, Uusi-Oukari M, Jänne OA (1997) Structure, organization and expression of the mouse ornithine decarboxylase antizyme gene. Biochem J 324:807–813

    CAS  PubMed  Google Scholar 

  • Liu GY, Liao YF, Hsu PC, Chang WH, Hsieh MC, Lin CY, Hour TC, Kao MC, Tsay GJ, Hung HC (2006) Antizyme, a natural ornithine decarboxylase inhibitor, induces apoptosis of haematopoietic cells through mitochondrial membrane depolarization and caspases’ cascade. Apoptosis 11:1773–1788

    Article  CAS  PubMed  Google Scholar 

  • Liu B, Sun H, Wang W, Li W, Yan Y, Chen S, Yang Y, Xu C, Xin J, Liu X (2009) Adenovirus vector-mediated upregulation of spermidine/spermineN1-acetyltransferase impairs human gastric cancer growth in vitro and in vivo. Cancer Sci 100:2126–2132

    Article  CAS  PubMed  Google Scholar 

  • Marshman E, Ottewell PD, Potten CS, Watson AJM (2001) Caspase activation during spontaneous and radiation-induced apoptosis in the murine intestine. J Pathol 195:285–292

    Article  CAS  PubMed  Google Scholar 

  • Martinez ME, O’Brien TG, Fultz KE, Babbar N, Yerushalmi H, Qu N, Guo Y, Boorman D, Einspahr J, Alberts DS, Gerner EW (2003) Pronounced reduction in adenoma recurrence associated with aspirin use and a polymorphism in the ornithine decarboxylase gene. Proc Natl Acad Sci USA 100:7859–7864

    Article  CAS  PubMed  Google Scholar 

  • Matsufuji S, Matsufuji T, Miyazaki Y, Murakami Y, Atkins JF, Gesteland RF, Hayashi S (1995) Autoregulatory frameshifting in decoding mammalian ornithine decarboxylase antizyme. Cell 80:51–60

    Article  CAS  PubMed  Google Scholar 

  • Meyskens J, McLaren CE, Pelot D et al (2008) Difluoromethylornithine plus sulindac for the prevention of sporadic colorectal adenomas: a randomized placebo-controlled, double-blind trial. Cancer Prev Res 1:32–38

    Article  CAS  Google Scholar 

  • Mitchell JL, Judd GG, Leyser A, Choe C (1998) Osmotic stress induces variation in cellular levels of ornithine decarboxylase-antizyme. Biochem J 329:453–459

    CAS  PubMed  Google Scholar 

  • Murakami Y, Matsufuji S, Miyazaki Y, Hayashi S (1994) Forced expression of antizyme abolishes ornithine decarboxylase activity, suppresses cellular levels of polyamines and inhibits cell growth. Biochem J 304:183–187

    CAS  PubMed  Google Scholar 

  • Perez-Leal O, Merali S (2012) Regulation of polyamine metabolism by translational control. Amino Acids 42:611–617

    Article  CAS  PubMed  Google Scholar 

  • Pietilä M, Lampinen A, Pellinen R, Alhonen L (2012) Inducible expression of antizyme 1 in prostate cancer cell lines after lentivirus mediated gene transfer. Amino Acids 42:559–564

    Article  PubMed  Google Scholar 

  • Shantz LM, Levin VA (2007) Regulation of ornithine decarboxylase during oncogenic transformation: mechanisms and therapeutic potential. Amino Acids 33:213–223

    Article  CAS  PubMed  Google Scholar 

  • Shin WW, Fong WF, Pang SF, Wong PC (1985) Limited blood-brain barrier transport of polyamines. J Neurochem 44:1056–1059

    Article  CAS  PubMed  Google Scholar 

  • Tosaka Y, Tanaka H, Yano Y, Masai K, Nozaki M, Yomogida K, Otani S, Nojima H, Nishimune Y (2000) Identification and characterization of testis specific ornithine decarboxylase antizyme (OAZ-t) gene: expression in haploid germ cells and polyamine-induced frameshifting. Genes Cells 5:265–276

    Article  CAS  PubMed  Google Scholar 

  • Wallace HM, Fraser AV (2004) Inhibitors of polyamine metabolism: review article. Amino Acids 26:353–365

    Article  CAS  PubMed  Google Scholar 

  • Wang X, Feith DJ, Welsh P, Coleman CS, Lopez C, Woster PM, O’Brien TG, Pegg AE (2007) Studies of the mechanism by which increased spermidine/spermine N1-acetyltransferase activity increases susceptibility to skin carcinogenesis. Carcinogenesis 28:2404–2411

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We thank Prof. Olli Jänne for providing the mutated mouse AZ1 gene construct and the rabbit polyclonal anti-AZ1 antibody. We also thank Ms. Marita Heikkinen, Sisko Juutinen, Arja Korhonen, Anne Karppinen and Tuula Reponen for their skillful technical assistance. This work was financially supported by the Academy of Finland.

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Authors and Affiliations

  1. Institute of Biomedicine, University of Eastern Finland, P.O.B 1627, 70211, Kuopio, Finland

    Marko Pietilä

  2. Department of Biotechnology and Molecular Medicine, A. I. Virtanen Institute for Molecular Sciences, Biocenter Kuopio, University of Eastern Finland, Kuopio, Finland

    Hiramani Dhungana, Anne Uimari & Leena Alhonen

  3. Institute of Clinical Medicine, Pathology and Forensic Medicine, Cancer Center of Eastern Finland, University of Eastern Finland, Kuopio, Finland

    Reijo Sironen

  4. Department of Pathology, Imaging Center, Kuopio University Hospital, Kuopio, Finland

    Reijo Sironen

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  1. Marko Pietilä
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  2. Hiramani Dhungana
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  4. Reijo Sironen
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  5. Leena Alhonen
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Correspondence to Marko Pietilä.

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Pietilä, M., Dhungana, H., Uimari, A. et al. Systemic overexpression of antizyme 1 in mouse reduces ornithine decarboxylase activity without major changes in tissue polyamine homeostasis. Transgenic Res 23, 153–163 (2014). https://doi.org/10.1007/s11248-013-9763-y

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  • DOI: https://doi.org/10.1007/s11248-013-9763-y

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