Aberrant splicing of folylpolyglutamate synthetase as a novel mechanism of antifolate resistance in leukemia

被引:72
作者
Stark, Michal [1 ]
Wichman, Chen [1 ]
Avivi, Irit [2 ,3 ]
Assaraf, Yehuda G. [1 ]
机构
[1] Technion Israel Inst Technol, Fred Wyszkowski Canc Res Lab, Dept Biol, IL-32000 Haifa, Israel
[2] Technion Israel Inst Technol, Dept Hematol, Rambam Med Ctr, IL-32000 Haifa, Israel
[3] Technion Israel Inst Technol, Rappaport Fac Med, IL-32000 Haifa, Israel
关键词
ACUTE LYMPHOBLASTIC-LEUKEMIA; GAMMA-GLUTAMATE SYNTHETASE; METHOTREXATE POLYGLUTAMATES; GENE-EXPRESSION; MOLECULAR-BASIS; MESSENGER-RNA; CELL; ACCUMULATION; 7-HYDROXYMETHOTREXATE; MULTIPLE;
D O I
10.1182/blood-2008-08-173799
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Folylpoly-gamma-gluatamate synthetase (FPGS) catalyzes the polyglutamylation and thus intracellular retention of folates and antifolates (eg, methotrexate; MTX) through the addition of multiple glutamate equivalents to their gamma-carboxyl residue. Since polyglutamylation of antifolates is crucial for their pharmacological activity in leukemia, loss of FPGS function results in decreased cellular levels of polyglutamylation-dependent antifolates and consequent drug resistance. Whereas resistance to pulse exposure to antifolates is frequently associated with loss of FPGS activity, the underlying molecular mechanism remains elusive. Here we explored the molecular basis of antifolate resistance in human MTX-resistant leukemia cell lines displaying marked loss of FPGS activity. We demonstrate that these MTX-resistant cells exhibit impaired splicing of FPGS mRNA based on intron retention and/or exon skipping, thereby resulting in loss of FPGS function due to premature translation termination. Furthermore, analysis of FPGS transcripts in blood or bone marrow specimens from patients with acute lymphoblastic leukemia revealed exon 12 skipping, both at diagnosis and at relapse, the latter of which occurs after high-dose MTX-containing chemotherapy. These results constitute the first demonstration of the loss of FPGS function via aberrant mRNA splicing, thereby resulting in loss of antifolate retention and drug resistance. The clinical ramifications of these novel findings are discussed. (Blood. 2009; 113: 4362-4369)
引用
收藏
页码:4362 / 4369
页数:8
相关论文
共 35 条
[1]   Molecular basis of antifolate resistance [J].
Assaraf, Yehuda G. .
CANCER AND METASTASIS REVIEWS, 2007, 26 (01) :153-181
[2]  
Barnes MJ, 1999, CLIN CANCER RES, V5, P2548
[3]  
BREITHAUPT H, 1982, CANCER TREAT REP, V66, P1733
[4]   The nonsense-mediated decay RNA surveillance pathway [J].
Chang, Yao-Fu ;
Imam, J. Saadi ;
Wilkinson, Miles E. .
ANNUAL REVIEW OF BIOCHEMISTRY, 2007, 76 :51-74
[5]   U2AF-homology motif interactions are required for alternative splicing regulation by SPF45 [J].
Corsini, Lorenzo ;
Bonnal, Sophie ;
Basquin, Jerome ;
Hothorn, Michael ;
Scheffzek, Klaus ;
Valcarcel, Juan ;
Sattler, Michael .
NATURE STRUCTURAL & MOLECULAR BIOLOGY, 2007, 14 (07) :620-U2
[6]   Resistance to Tomudex (ZD1694): Multifactorial in human breast and colon carcinoma cell lines [J].
Drake, JC ;
Allegra, CJ ;
Moran, RG ;
Johnston, PG .
BIOCHEMICAL PHARMACOLOGY, 1996, 51 (10) :1349-1355
[7]   TEMPORARY REMISSIONS IN ACUTE LEUKEMIA IN CHILDREN PRODUCED BY FOLIC ACID ANTAGONIST, 4-AMINOPTEROYL-GLUTAMIC ACID (AMINOPTERIN) [J].
FARBER, S ;
DIAMOND, LK ;
MERCER, RD ;
SYLVESTER, RF ;
WOLFF, JA .
NEW ENGLAND JOURNAL OF MEDICINE, 1948, 238 (23) :787-793
[8]   Disparate mechanisms of antifolate resistance provoked by methotrexate and its metabolite 7-hydroxymethotrexate in leukemia cells: implications for efficacy of methotrexate therapy [J].
Fotoohi, K ;
Jansen, G ;
Assaraf, YG ;
Rothem, L ;
Stark, M ;
Kathmann, I ;
Gregorczyk, J ;
Peters, GJ ;
Albertioni, F .
BLOOD, 2004, 104 (13) :4194-4201
[9]   In adults with standard-risk acute lymphoblastic leukemia, the greatest benefit is achieved from a matched sibling allogeneic transplantation in first complete remission, and an autologous transplantation is less effective than conventional consolidation/maintenance chemotherapy in all patients: final results of the International ALL Trial (MRC UKALL XII/ECOG E2993) [J].
Goldstone, Anthony H. ;
Richards, Susan M. ;
Lazarus, Hillard M. ;
Tallman, Martin S. ;
Buck, Georgina ;
Fielding, Adele K. ;
Burnett, Alan K. ;
Chopra, Raj ;
Wiernik, Peter H. ;
Foroni, Letizia ;
Paietta, Elisabeth ;
Litzow, Mark R. ;
Marks, David I. ;
Durrant, Jill ;
McMillan, Andrew ;
Franklin, Ian M. ;
Luger, Selina ;
Ciobanu, Niculae ;
Rowe, Jacob M. .
BLOOD, 2008, 111 (04) :1827-1833
[10]   Quality control of eukaryotic mRNA: safeguarding cells from abnormal mRNA function [J].
Isken, Olaf ;
Maquat, Lynne E. .
GENES & DEVELOPMENT, 2007, 21 (15) :1833-1856