#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Diverse asparagine synthetase expression in lymphoid blasts is not relevant to the sensitivity to L-Asparaginase


Authors: I. Heřmanová;  J. Trka;  J. Stárková
Authors‘ workplace: Klinika dětské hematologie a onkologie ;  UK 2. LF a FN v Motole, Praha ;  CLIP – Childhood Leukemia Investigation Prague
Published in: Transfuze Hematol. dnes,16, 2010, No. 3, p. 133-140.
Category: Comprehensive Reports, Original Papers, Case Reports

Overview

Acute lymphoblastic leukaemia (ALL), the most common haematological malignancy in childhood, is treated by combined chemotherapy, which includes the enzyme L-Asparaginase (L-Asp). The cytotoxic effect of L-Asp consists in its ability to deplete extracellular asparagine and glutamine. The sensitivity of primary ALL cells to this depletion is traditionally explained by decreased activity of glutamine-dependent enzyme asparagine synthetase (ASNS). Despite the fact that increased ASNS level was indeed shown to be connected with L-Asp resistance, the exact relationship between ASNS expression and L-Asp sensitivity has not been elucidated so far. The gene expression and ASNS protein content was evaluated in 4 leukemic cell lines: Nalm6 (TEL/PDGFRB[+]); RS4;11 (MLL/AF4[+]); REH (TEL/AML1[+]) and UOCB6 (TEL/AML1[+]). ASNS protein levels reflected ASNS mRNA levels and these correlated negatively with L-Asp sensitivity. UOCB6 as the most resistant cell line had the highest expression of ASNS, followed by Nalm6, REH and RS4;11. Detection of protein content in primary ALL blasts was not possible due to significantly lower ASNS gene expression compared to cell lines. Gradient knock-down was performed in 2 ALL cell lines: REH with intermediate basal expression and RS4;11 with very low basal expression. A gradual silencing of ASNS gene in REH cell line led to gradual increase of L-Asp sensitivity. The reduction of ASNS did not potentiate L-Asp cytotoxicity in RS4;11 cell line. Our data demonstrate that in cells with very low ASNS expression, as shown in primary ALL blasts of various subtypes, the difference in ASNS levels is not relevant to the sensitivity to L-Asp. We suppose that glutamate dehydrogenase (GDH) is a new player in the response to cytotoxic effect of L-Asp. Silencing of GDH gene in TEL/AML1[+] REH cell line increased sensitivity to L-Asp. Furthermore, we suggest a relationship between ASNS and GDH based on our observations of increased GDH expression in cells with silenced ASNS gene.

Key words:
acute lymphoblastic leukaemia, L-Asparaginase, asparagine synthetase


Sources

1. Pui CH, Evans WE. Acute lymphoblastic leukemia. N Engl J Med 1998; 339: 605-615.

2. Pui CH, Evans WE. Treatment of acute lymphoblastic leukemia. N Engl J Med 2006; 354: 166-178.

3. Jaffe N, Traggis D, Das L, et al. L-Asparaginase in treatment of neoplastic diseases in children. Cancer Res 1971; 31: 934-949.

4. Sutow WW, Garacia F, Starling KA, Williams TE, Lane DM, Gehan EA. L-Asparaginase therapy in children with advanced leukemia. Cancer 1971; 28: 819-824.

5. Tallal L, Tan C, Oettgen H, et al. E. coli L-asparaginase in the treatment of leukemia and solid tumors in 131 children. Cancer 1970; 25: 306-320.

6. Chakrabarti R, Schuster SM. L-Asparaginase. Perspectives on the mechanisms of action and resistance. Int J Pediatric Hematol Oncol 1996; 4: 597-611.

7. Muller HJ, Boos J. Use of L-asparaginase in childhood ALL. Crit Rev Oncol Hematol 1998; 28: 97-113.

8. Chen H, Pan YX, Dudenhausen EE, et al. Amino acid deprivation induces the transcription rate of the human asparagine synthetase gene through a timed program of expression and promoter binding of nutrient-responsive bZIP transcription factors as well as localized histone acetylation. J Biol Chem 2004; 279: 50829-50839.

9. Kilberg MS, Pan YX, Chen H, et al. Nutritional control of gene expression: How mammalian cells respond to amino acid limitation. Annu Rev Nutr 2005; 25: 59-85.

10. Asselin BL, Kurtzburg J. Asparaginase. In: Pui C-H, editor. Treatment of acute leukemias. Totawa, NJ: Humana Press; 2003: 365-379.

11. Rizzari C. Asparaginase treatment. In: Pui C-H, editor. Treatment of acute leukemias. Towata, NJ: Humana Press; 2003. 381-391.

12. Richards NG, Kilberg MS. Asparagine synthetase chemotherapy. Ann Rev Biochem 2006; 75: 629-654.

13. Richards NG, Schuster SM. Mechanistic issues in asparagine synthetase catalysis. Adv Enzymol Relat Areas Mol Biol 1998; 72: 145-198.

14. Kiriyama Y, Kubota M, Takimoto T, et al. Biochemical characterization of U937 cells resistant to L-asparaginase: The role of asparagine synthetase. Leukemia 1989; 3:294–7.

15. Su N, Pan YX, Zhou M, Harvey RC, Hunger SP, Kilberg MS. Correlation between asparaginase sensitivity and asparagine synthetase protein content, but no mRNA, in acute lymphoblastic cell lines. Pediatr Blood Cancer 2008; 50: 274-279.

16. Wang L, Hiebert SW. TEL contacts multiple co-repressors and specifically associates with histone deacetylase-3. Oncogene 2001; 20: 3761-3725.

17. Zuna J. The role of TEL and AML1 genes in the pathogenesis of hematologic malignancies. Cas Lek Cesk 2001; 140: 131-137.

18. Zuna J, Hrusak O, Kalinova M, Muzikova K, Stary J, Trka J. TEL/AML1 positivity in childhood ALL: average or better prognosis? Czech Paediatric Haematology Working Group. Leukemia 1999, 13: 22-24.

19. Ramakers-van Woerden NL, Pieters R, Loonen AH, et al. TEL/AML1 gene fusion is related to in vitro drug sensitivity for L-Asparaginase in childhood acute lymphoblastic leukemia. Blood 2000; 96: 1094-1099.

20. Krejci O, Starkova J, Otova B, Madzo J, Kalinova M, Hrusak O, Trka J. Upregulation of asparagine synthetase fails to avert cell cycle arrest induced by L-Asparaginase in TEL/AML1-positive leukaemic cells. Leukemia 2004; 18: 434-441.

21. Stams WA, den Boer ML, Beverloo HB, et al. Sensitivity to l-asparaginase is not associated with expression levels of asparagine synthetase in t(12,21)+ pediatric ALL. Blood 2003; 101: 2743-274.

22. Fine BM, Kaspers GJL, Ho M, Loonen AH, Boxer LM. A genome wide view of the in vitro response to l-asparaginase in acute lymphoblastic leukemia. Cancer Res 2005; 65: 291-299.

23. O’Toole SA, Sheppard BL, McGuinness EPJ, Gleeson NC, Yoneda M, Bonnar JJ. The MTS assay as an indicator of chemosensitivity/resistance in malignant gynaecological tumours. Cancer Detection and Prevention 2003; 27: 47-54

24. Scherf U, Ross DT, Waltham M, et al. A gene expression database for the molecular pharmacology of cancer. Nat Genet 2000; 24: 236-244.

25. Starková J, Krejčí O, Otová B, et al. TEL/AML1 spustením metabolického stresu zvyšuje citlivosť leukemických buniek na L-Asparaginázu. Transfuze Hematol dnes 2006; 12: 70-75.

26. Hutson RG, Kitoh T, Amador DAM, Cosic S, Schuster SM, Kilberg MS. Amino acid control of asparagine synthetase : relation to asparaginase resistance in human leukemia cells. Am J Physiol 1997; 272: C1691-C1699.

27. Prager MD, Bachynsky N. Asparagine synthetase in asparaginase resistant and susceptible mouse lymphomas. Biochem Biophys Res Commun 1968; 31: 43-47.

28. Worton KS, Kerbel RS, Andrulis IL. Hypomethylation and reactivation of the asparagine synthetase gene induced by L-asparaginase and ethyl methanesulfonate. Cancer Res 1991; 51: 985-989.

29. Martin JK, Sun W, Maraga D, Schuster SM, Wylie DE. An investigation into the mechanism of L-asparaginase resistance in L5178Y murine leukemia cells. Amino Acids 1993; 5: 51-69.

30. Aslanian AM, Fletcher BS, Kilberg MS. Asparagine synthetase expression alone is sufficient to induce L-Asparaginase resistance in MOLT-4 human leukemia cells. Biochem J 2001; 357: 321-328.

31. Haskell CM, Canellos GP. L-Asparaginase resistance in human leukemia-asparagine synthetase. Biochem Pharm 1969; 18: 2578-2580.

32. Stams WAG, den Boer ML, Holleman A, et al. Asparagine synthetase expression is linked with L-Asparaginase resistance in TEL-AML-negative but not TEL-AML-positive pediatric acute lymphoblastic leukemia. Blood 2005; 105: 4223-4225.

33. Patel N, Krishnan S, Offman MN, et al. A dyad of lymphoblastic lysosomal cysteine proteases degrades the antileukemic drug L-asparaginase. J Clin Invest 2009; 119: 1964-1973.

34. Estes DA, Lovato DM, Khawaja HM, Winter SS, Larson RS. Genetic alterations determine chemotherapy resistance in childhood T-ALL: modeling in stage-specific cell lines and correlation with diagnostic patient samples. Br J Haematol 2007; 139: 20-30.

Labels
Haematology Internal medicine Clinical oncology
Topics Journals
Login
Forgotten password

Enter the email address that you registered with. We will send you instructions on how to set a new password.

Login

Don‘t have an account?  Create new account

#ADS_BOTTOM_SCRIPTS#