Ukr.Biochem.J. 2016; Volume 88, Issue 2, Mar-Apr, pp. 45-55

doi: https://doi.org/10.15407/ubj88.02.045

Effects of the combined arginase and canavanine treatment on leukemic cells in vitro and in vivo

O. I. Vovk1, O. I. Chen1,2, N. I. Igumentseva1, O. Yu. Senchuk1,
M. L. Barska1, N. O. Sybirna1,2, O. V. Stasyk1

 1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
2Ivan Franko National University of Lviv, Ukraine;
e-mail: stasyk@cellbiol.lviv.ua

It was previously demonstrated in in vitro experiments that canavanine (Cav), a natural toxic arginine analogue of plant origin, is a promising candidate for augmenting the antineoplastic effects of arginine starvation. We demonstrated herein that recombinant human arginase, an arginine degrading enzyme, abrogated growth and significantly increased Cav cytotoxicity toward cultured L1210 murine leukemic cells. Cav co-treatment further reduced cells viability in a time-dependent manner and significantly promoted apoptosis induction. In the pilot study we also evaluated for the first time the potential toxicity of the combined arginine deprivation and Cav treatment in healthy mice. Administration of Cav alone or in combination with pegylated cobalt-containing human arginase (Co-hARG) did not evoke any apparent toxic effects in these animals, with no significant behavioural and survival changes after several weeks of the treatment. The therapeutic effects of the combination of Co-hARG and Cav were provisionally evaluated on the highly aggressive murine L1210 leukemia, which is semi-sensitive to arginine deprivation as a monotreatment. Combination of two drugs did not result in significant prolongation of the survival of leukemia-bearing mice. Thus, we have shown that the proposed combinational treatment is rather non-toxic for the animals. It has to be further evaluated in animal studies with alternative tumor models and/or drug doses and treatment modalities.

Keywords: , , ,


References:

  1. Ott PA, Carvajal RD, Pandit-Taskar N, Jungbluth AA, Hoffman EW, Wu BW, Bomalaski JS, Venhaus R, Pan L, Old LJ, Pavlick AC, Wolchok JD. Phase I/II study of pegylated arginine deiminase (ADI-PEG 20) in patients with advanced melanoma. Invest New Drugs. 2013 Apr;31(2):425-34. PubMed, PubMedCentral, CrossRef
  2. Yau T, Cheng PN, Chan P, Chen L, Yuen J, Pang R, Fan ST, Wheatley DN, Poon RT. Preliminary efficacy, safety, pharmacokinetics, pharmacodynamics and quality of life study of pegylated recombinant human arginase 1 in patients with advanced hepatocellular carcinoma. Invest New Drugs. 2015 Apr;33(2):496-504.  PubMed, CrossRef
  3. Qiu F, Huang J, Sui M. Targeting arginine metabolism pathway to treat arginine-dependent cancers. Cancer Lett. 2015 Aug 1;364(1):1-7. Review. PubMed, CrossRef
  4. Stasyk OV, Boretsky YR, Gonchar MV, Sibirny AA. Recombinant arginine-degrading enzymes in metabolic anticancer therapy and bioanalytics. Cell Biol Int. 2015 Mar;39(3):246-52. Review. PubMed, CrossRef
  5. Phillips MM, Sheaff MT, Szlosarek PW. Targeting arginine-dependent cancers with arginine-degrading enzymes: opportunities and challenges. Cancer Res Treat. 2013 Dec;45(4):251-62. Review. PubMed, PubMedCentral, CrossRef
  6. Feun LG, Kuo MT, Savaraj N. Arginine deprivation in cancer therapy. Curr Opin Clin Nutr Metab Care. 2015 Jan;18(1):78-82. Review. PubMed, CrossRef
  7. Rosenthal GA. Plant nonprotein amino and imino acids: biological, biochemical, and toxicological properties, Academic Press, New York NY, 1982; 95.
  8. Blind PJ, Waldenström A, Berggren D, Ronquist G. Antitumour effect of L-2,4 diaminobutyric acid on a hepatoma cell line. Anticancer Res. 2000 Nov-Dec;20(6B):4275-8. PubMed
  9. Rosenthal G.A., In: Singh B.K., Flores H.E., Shannon J.C. (eds) Biosynthesis and molecular regulation of aminoacids in plants, American society of plant physiologists, Rockville, MD, 1992; 249.
  10. Umans JG, Samsel RW. L-canavanine selectively augments contraction in aortas from endotoxemic rats. Eur J Pharmacol. 1992 Jan 21;210(3):343-6. PubMed, CrossRef
  11. Green MH, Brooks TL, Mendelsohn J, Howell SB. Antitumor activity of L-canavanine against L1210 murine leukemia. Cancer Res. 1980 Mar;40(3):535-7. PubMed
  12. Thomas DA, Rosenthal GA, Gold DV, Dickey K. Growth inhibition of a rat colon tumor by L-canavanine. Cancer Res. 1986 Jun;46(6):2898-903. PubMed
  13. Swaffar DS, Ang CY, Desai PB, Rosenthal GA. Inhibition of the growth of human pancreatic cancer cells by the arginine antimetabolite L-canavanine. Cancer Res. 1994 Dec 1;54(23):6045-8. PubMed
  14. Green MH, Ward JF. Enhancement of human tumor cell killing by L-canavanine in combination with gamma-radiation. Cancer Res. 1983 Sep;43(9):4180-2. PubMed
  15. Swaffar DS, Ang CY. Growth inhibitory effect of L-canavanine against MIA PaCa-2 pancreatic cancer cells is not due to conversion to its toxic metabolite canaline. Anticancer Drugs. 1999 Jan;10(1):113-8. PubMed, CrossRef
  16. Vynnytska BO, Mayevska OM, Kurlishchuk YV, Bobak YP, Stasyk OV. Canavanine augments proapoptotic effects of arginine deprivation in cultured human cancer cells. Anticancer Drugs. 2011 Feb;22(2):148-57.  PubMed, CrossRef
  17. Chen O, Kavalets B, Barska M, Lyniv L, Vovk O, Sybirna N Stasyk O. Effect of combinational arginase and canavanine treatment on normal human peripheral blood lymphocytes in vitro. Curr Issues Pharm Med Sci. 2013; 26(4): 385-389. CrossRef
  18. Chen O, Kavaletz B., Lyniv L, Barska M, Vovk O, Sybirna N, Stasyk O. Effect of combinational arginine deprivation-based enzymotherapy (CADE) with canavanine on viability of human leukemic cells in vitro/ International symposium on Cell Biology jointly with 3rd Ukrainian Congresss for Cell Biology. Yalta, Ukraine, 2012. P.99.
  19. Chen O., Barska M., Sybirna N., Stasyk O. Development and evaluation of HPLC method for arginine assay in biological samples. Visnyk Lviv Univ. Series Biology. 2012;60:73-82.
  20. Peterson GL. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346-56. PubMed, CrossRef
  21. Menshikova VV. Laboratory methods of investigation in the clinic. M.: Medicina, 1987. 190 p. (in Russian).
  22. Caraway WT. A stable starch substrate for the determination of amylase in serum and other body fluids. Am J Clin Pathol. 1959 Jul;32(1):97-9. PubMed
  23. Marsh WH, Fingerhut B, Miller H. Automated and manual direct methods for the determination of blood urea. Clin Chem. 1965 Jun;11:624-7. PubMed
  24. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Biotechnology. 1992;24:145-9. PubMed
  25. Stone EM, Glazer ES, Chantranupong L, Cherukuri P, Breece RM, Tierney DL, Curley SA, Iverson BL, Georgiou G. Replacing Mn(2+) with Co(2+) in human arginase i enhances cytotoxicity toward l-arginine auxotrophic cancer cell lines. ACS Chem Biol. 2010 Mar 19;5(3):333-42. PubMed, PubMedCentral, CrossRef
  26. Glazer ES, Stone EM, Zhu C, Massey KL, Hamir AN, Curley SA. Bioengineered human arginase I with enhanced activity and stability controls hepatocellular and pancreatic carcinoma xenografts. Transl Oncol. 2011 Jun;4(3):138-46. PubMed, PubMedCentral, CrossRef
  27. Hall AP, Elcombe CR, Foster JR, Harada T, Kaufmann W, Knippel A, Küttler K, Malarkey DE, Maronpot RR, Nishikawa A, Nolte T, Schulte A, Strauss V, York MJ. Liver hypertrophy: a review of adaptive (adverse and non-adverse) changes–conclusions from the 3rd International ESTP Expert Workshop. Toxicol Pathol. 2012 Oct;40(7):971-94.  PubMed, CrossRef
  28. Bence AK, Crooks PA. The mechanism of L-canavanine cytotoxicity: arginyl tRNA synthetase as a novel target for anticancer drug discovery. J Enzyme Inhib Med Chem. 2003 Oct;18(5):383-94. PubMed, CrossRef

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