Detailed Functional and Proteomic Characterization of Fludarabine Resistance in Mantle Cell Lymphoma Cells.
Mantle cell lymphoma (MCL) is a chronically relapsing aggressive type of B-cell non-Hodgkin lymphoma considered incurable by currently used treatment approaches. Fludarabine is a purine analog clinically still widely used in the therapy of relapsed MCL. Molecular mechanisms of fludarabine resistance have not, however, been studied in the setting of MCL so far. We therefore derived fludarabine-resistant MCL cells (Mino/FR) and performed their detailed functional and proteomic characterization compared to the original fludarabine sensitive cells (Mino). We demonstrated that Mino/FR were highly cross-resistant to other antinucleosides (cytarabine, cladribine, gemcitabine) and to an inhibitor of Bruton tyrosine kinase (BTK) ibrutinib. Sensitivity to other types of anti-lymphoma agents was altered only mildly (methotrexate, doxorubicin, bortezomib) or remained unaffacted (cisplatin, bendamustine). The detailed proteomic analysis of Mino/FR compared to Mino cells unveiled over 300 differentially expressed proteins. Mino/FR were characterized by the marked downregulation of deoxycytidine kinase (dCK) and BTK (thus explaining the observed crossresistance to antinucleosides and ibrutinib), but also by the upregulation of several enzymes of de novo nucleotide synthesis, as well as the up-regulation of the numerous proteins of DNA repair and replication. The significant upregulation of the key antiapoptotic protein Bcl-2 in Mino/FR cells was associated with the markedly increased sensitivity of the fludarabine-resistant MCL cells to Bcl-2-specific inhibitor ABT199 compared to fludarabine-sensitive cells. Our data thus demonstrate that a detailed molecular analysis of drug-resistant tumor cells can indeed open a way to personalized therapy of resistant malignancies.
Institute of Pathological Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
Lucie
L
Lorkova
Thermo Fisher Scientific, Bremen, Germany.
Michaela
M
Scigelova
Thermo Fisher Scientific, Bremen, Germany.
Tabiwang Ndipanquang
TN
Arrey
Institute of Pathological Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
Ondrej
O
Vit
Institute of Pathological Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
Jana
J
Pospisilova
Institute of Pathological Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
Eliska
E
Doktorova
Institute of Pathological Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic; First Department of Medicine-Department of Hematology, General University Hospital and Charles University in Prague, Prague, Czech Republic.
Magdalena
M
Klanova
Institute of Pathological Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
Mahmudul
M
Alam
Institute of Pathological Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic; First Department of Medicine-Department of Hematology, General University Hospital and Charles University in Prague, Prague, Czech Republic.
Petra
P
Vockova
Institute of Pathological Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
Bokang
B
Maswabi
Institute of Pathological Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic; First Department of Medicine-Department of Hematology, General University Hospital and Charles University in Prague, Prague, Czech Republic.
Pavel
P
Klener
Institute of Pathological Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic; Institute of Hematology and Blood Transfusion, Prague, Czech Republic.
Jiri
J
Petrak
D000970:Antineoplastic Agents / Q000494:pharmacology*; D014408:Biomarkers, Tumor / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods; D019008:Drug Resistance, Neoplasm; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D020522:Lymphoma, Mantle-Cell / Q000188:drug therapy / Q000378:metabolism*; D008297:Male; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods; D014407:Tumor Cells, Cultured; D014740:Vidarabine / Q000031:analogs & derivatives* / Q000494:pharmacology
D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D014740:Vidarabine; C024352:fludarabine