Genetic alterations determine chemotherapy resistance in childhood T‐ALL: modelling in stage‐specific cell lines and correlation with diagnostic patient samples

Acquired drug resistance eventually leads to treatment failure in T‐cell acute lymphoblastic leukaemia (T‐ALL). Immunophenotypic and cytogenetic heterogeneities within T‐ALL influence susceptibility to cytotoxic therapy, and little is known about the mechanisms of drug resistance at specific stages of T‐cell ontogeny. We developed tolerance to therapeutic concentrations of daunorubicin (DNR) and l‐asparaginase (l‐asp) in Jurkat (CD1a−, sCD3+) and Sup T1 (CD1a+, sCD3−) cell lines, having respective ‘mature’ and ‘cortical’ stages of developmental arrest. DNR resistant cells acquired multidrug resistance: 310‐fold increased resistance to vincristine (VCR) and a 120‐fold increased resistance to prednisolone (PRED). Microarray analysis identified upregulation of asparagine synthetase (ASNS) and argininosuccinate synthase 1 (ASS1) to cell lines with acquired resistance to l‐asp, and in the case of DNR, upregulation of ATP‐binding cassette B1 (ABCB1). Suppression of ABCB1, ASNS and ASS1 by RNA interference revealed their functional relevance to acquired drug resistance. Expression profiling of these genes in 80 T‐ALL patients showed correlation with treatment response. This study expands the pool of available drug resistant cell lines having cortical and mature stages of developmental arrest, introduces three new drug resistant T‐ALL cell lines, and identifies gene interactions leading to l‐asp and DNR resistance.

[1]  F. Behm,et al.  Bone marrow-derived stromal cells prevent apoptotic cell death in B-lineage acute lymphoblastic leukemia. , 1992, Blood.

[2]  M. Tiirikainen,et al.  INITIAL P-GLYCOPROTEIN EXPRESSION IN CHILDHOOD ACUTE LYMPHOBLASTIC LEUKEMIA: NO EVIDENCE OF PROGNOSTIC IMPACT IN FOLLOW-UP , 2001, Pediatric hematology and oncology.

[3]  C. Bloomfield,et al.  Value of immunophenotype in intensively treated adult acute lymphoblastic leukemia: cancer and leukemia Group B study 8364. , 1999, Blood.

[4]  B. Dörken,et al.  In vitro susceptibility to dexamethasone- and doxorubicin-induced apoptotic cell death in context of maturation stage, responsiveness to interleukin 7, and early cytoreduction in vivo in childhood T-cell acute lymphoblastic leukemia. , 2002, Blood.

[5]  L. Boxer,et al.  A genome-wide view of the in vitro response to l-asparaginase in acute lymphoblastic leukemia. , 2005, Cancer research.

[6]  J. Robert,et al.  Mitochondrial localization and activity of P-glycoprotein in doxorubicin-resistant K562 cells. , 2006, Biochemical pharmacology.

[7]  Z. Darżynkiewicz.  Differential staining of DNA and RNA in intact cells and isolated cell nuclei with acridine orange. , 1990, Methods in cell biology.

[8]  M. Tiirikainen,et al.  Multiple drug resistance mediated by P-glycoprotein is not a major factor in a slow response to therapy in childhood ALL. , 1998, Pediatric hematology and oncology.

[9]  Cheng Cheng,et al.  Gene-expression patterns in drug-resistant acute lymphoblastic leukemia cells and response to treatment. , 2004, The New England journal of medicine.

[10]  H. Miyachi,et al.  MDR1 (multidrug resistance) gene expression in adult acute leukemia: correlations with blast phenotype. , 1993, International journal of hematology.

[11]  J. Philippé,et al.  The Combined Analysis of P-Glycoprotein Expression and Activity Predicts Outcome in Childhood Acute Lymphoblastic Leukemia , 2003, Pediatric hematology and oncology.

[12]  M. Prager,et al.  Asparagine synthetase in normal and malignant tissues: correlation with tumor sensitivity to asparaginase. , 1968, Archives of biochemistry and biophysics.

[13]  H. Drexler,et al.  Heterogeneity of T-acute lymphoblastic leukemia (T-ALL) cell lines: suggestion for classification by immunophenotype and T-cell receptor studies. , 1999, Leukemia research.

[14]  M. Amylon,et al.  Intensive high-dose asparaginase consolidation improves survival for pediatric patients with T cell acute lymphoblastic leukemia and advanced stage lymphoblastic lymphoma: a Pediatric Oncology Group study , 1999, Leukemia.

[15]  M. Relling,et al.  Childhood Acute Lymphoblastic Leukaemia , 2007 .

[16]  M. Prager,et al.  Asparagine synthetase in asparaginase resistant and susceptible mouse lymphomas. , 1968, Biochemical and biophysical research communications.

[17]  H. Sather,et al.  Response of children with high-risk acute lymphoblastic leukemia treated with and without cranial irradiation: a report from the Children's Cancer Group. , 1998, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[18]  MDR1 protein expression is an independent predictor of complete remission in newly diagnosed adult acute lymphoblastic leukemia. , 2002, Blood.

[19]  R. Pieters,et al.  In vitro drug resistance profiles of adult versus childhood acute lymphoblastic leukaemia , 2000, British journal of haematology.

[20]  T. Hongo,et al.  In vitro drug sensitivity testing can predict induction failure and early relapse of childhood acute lymphoblastic leukemia. , 1997, Blood.

[21]  M. D. Boer,et al.  Relation between age, immunophenotype and in vitro drug resistance in 395 children with acute lymphoblastic leukemia–implications for treatment of infants , 1998, Leukemia.

[22]  Richard S Larson,et al.  Identification of genomic classifiers that distinguish induction failure in T-lineage acute lymphoblastic leukemia: a report from the Children's Oncology Group. , 2007, Blood.

[23]  R L Juliano,et al.  A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. , 1976, Biochimica et biophysica acta.

[24]  A Orfao,et al.  Proposals for the immunological classification of acute leukemias. European Group for the Immunological Characterization of Leukemias (EGIL). , 1995, Leukemia.

[25]  B. Seed,et al.  A PCR primer bank for quantitative gene expression analysis. , 2003, Nucleic acids research.

[26]  D. Marquardt,et al.  Multiple mechanisms of adriamycin resistance in the human leukemia cell line CCRF-CEM. , 1989, Biochemical pharmacology.

[27]  E. Lander,et al.  Gene expression signatures define novel oncogenic pathways in T cell acute lymphoblastic leukemia. , 2002, Cancer cell.

[28]  H. Sather,et al.  Asparaginase Antibody and Asparaginase Activity in Children With Higher-Risk Acute Lymphoblastic Leukemia: Children's Cancer Group Study CCG-1961 , 2004, Journal of pediatric hematology/oncology.

[29]  E. Paietta.  Proposals for the immunological classification of acute leukemias. , 1995, Leukemia.

[30]  Katrin Hoffmann,et al.  Gene expression levels assessed by oligonucleotide microarray analysis and quantitative real-time RT-PCR – how well do they correlate? , 2005, BMC Genomics.

[31]  K. Oka,et al.  Induction of multidrug resistance in MOLT-4 cells by anticancer agents is closely related to increased expression of functional P-glycoprotein and MDR1 mRNA , 2002, Cancer Chemotherapy and Pharmacology.

[32]  M. Kilberg,et al.  Asparagine synthetase expression alone is sufficient to induce l-asparaginase resistance in MOLT-4 human leukaemia cells. , 2001, The Biochemical journal.

[33]  H. Mikawa,et al.  Biochemical characterization of U937 cells resistant to L-asparaginase: the role of asparagine synthetase. , 1989, Leukemia.

[34]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[35]  X. Li,et al.  Sensitization of multidrug-resistant human leukemia cells with MDR1-targeted antisense and inhibition of drug-mediated MDR1 induction , 1997, Leukemia.

[36]  M. D. Boer,et al.  Mechanisms of cellular anthracycline resistance in childhood acute leukemia , 1998, Leukemia.

[37]  P. Sonneveld.  Multidrug resistance in haematological malignancies , 2000, Journal of internal medicine.

[38]  S. Arfin,et al.  Properties of asparagine synthetase in asparagine-independent variants of Jensen rat sarcoma cells induced by 5-azacytidine , 1983, Molecular and cellular biology.

[39]  J. Burchenal,et al.  Treatment of acute lymphoblastic leukemia. , 1972, Annual review of medicine.

[40]  C. Vinson,et al.  Regulation of asparagine synthetase gene transcription by the basic region leucine zipper transcription factors ATF5 and CHOP , 2005, Biological chemistry.

[41]  J. Philippé,et al.  P‐glycoprotein is an independent prognostic factor predicting relapse in childhood acute lymphoblastic leukaemia: results of a 6‐year prospective study , 1999, British journal of haematology.

[42]  M. D. Boer,et al.  Patient stratification based on prednisolone-vincristine-asparaginase resistance profiles in children with acute lymphoblastic leukemia. , 2003, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[43]  F. de Longueville,et al.  Expression profiling of ATP-binding cassette transporters in childhood T-cell acute lymphoblastic leukemia , 2006, Molecular Cancer Therapeutics.

[44]  W. T. Beck.  Vinca alkaloid-resistant phenotype in cultured human leukemic lymphoblasts. , 1983, Cancer treatment reports.

[45]  N. Heerema,et al.  Biology and treatment of childhood T-lineage acute lymphoblastic leukemia. , 1998, Blood.

[46]  Z. Li,et al.  Epigenetic changes in the repression and induction of asparagine synthetase in human leukemic cell lines , 2005, Leukemia.

[47]  J. Philippé,et al.  Expression of the Multidrug Transporter P-glycoprotein is Highly Correlated with Clinical Outcome in Childhood Acute Lymphoblastic Leukemia: Results of a Long-Term Prospective Study , 2002, Leukemia & lymphoma.