The enzymatic activity of 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase is enhanced by NPM-ALK: new insights in ALK-mediated pathogenesis and the treatment of ALCL.
Anaplastic large cell lymphoma represents a subset of neoplasms caused by translocations that juxtapose the anaplastic lymphoma kinase (ALK) to dimerization partners. The constitutive activation of ALK fusion proteins leads to cellular transformation through a complex signaling network. To elucidate the ALK pathways sustaining lymphomagenesis and tumor maintenance, we analyzed the tyrosine-kinase protein profiles of ALK-positive cell lines using 2 complementary proteomic-based approaches, taking advantage of a specific ALK RNA interference (RNAi) or cell-permeable inhibitors. A well-defined set of ALK-associated tyrosine phosphopeptides, including metabolic enzymes, kinases, ribosomal and cytoskeletal proteins, was identified. Validation studies confirmed that vasodilator-stimulated phosphoprotein and 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/inosine monophosphate cyclohydrolase (ATIC) associated with nucleophosmin (NPM)-ALK, and their phosphorylation required ALK activity. ATIC phosphorylation was documented in cell lines and primary tumors carrying ALK proteins and other tyrosine kinases, including TPR-Met and wild type c-Met. Functional analyses revealed that ALK-mediated ATIC phosphorylation enhanced its enzymatic activity, dampening the methotrexate-mediated transformylase activity inhibition. These findings demonstrate that proteomic approaches in well-controlled experimental settings allow the definition of informative proteomic profiles and the discovery of novel ALK downstream players that contribute to the maintenance of the neoplastic phenotype. Prediction of tumor responses to methotrexate may justify specific molecular-based chemotherapy.
Center for Experimental Research and Medical Studies (CERMS), University of Torino, Via Santena 5, 10126 Torino, Italy.
Francesco E
FE
Boccalatte
Claudia
C
Voena
Chiara
C
Riganti
Amalia
A
Bosia
Lucia
L
D'Amico
Ludovica
L
Riera
Mangeng
M
Cheng
Bruce
B
Ruggeri
Ole N
ON
Jensen
Valerie L
VL
Goss
Kimberly
K
Lee
Julie
J
Nardone
John
J
Rush
Roberto D
RD
Polakiewicz
Michael J
MJ
Comb
Roberto
R
Chiarle
Giorgio
G
Inghirami
D000595:Amino Acid Sequence; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D002227:Carbazoles / Q000494:pharmacology; D015815:Cell Adhesion Molecules / Q000378:metabolism; D045744:Cell Line, Tumor / Q000187:drug effects / Q000378:metabolism; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D020869:Gene Expression Profiling; D006801:Humans; D019877:Hydroxymethyl and Formyl Transferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007191:Indazoles / Q000494:pharmacology; D017728:Lymphoma, Large-Cell, Anaplastic / Q000188:drug therapy / Q000201:enzymology* / Q000473:pathology; D008727:Methotrexate / Q000494:pharmacology; D008840:Microfilament Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009709:Nucleotide Deaminases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010671:Phenylurea Compounds / Q000494:pharmacology; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000032:analysis; D025941:Protein Interaction Mapping; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014158:Transcription, Genetic; D000098662:Vasodilator-Stimulated Phosphoprotein
D000964:Antimetabolites, Antineoplastic; C540056:CEP 11988; C540057:CEP 14083; D002227:Carbazoles; D015815:Cell Adhesion Molecules; D007191:Indazoles; D008840:Microfilament Proteins; D009097:Multienzyme Complexes; D009363:Neoplasm Proteins; D010671:Phenylurea Compounds; D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; C113120:inosine monophosphate synthase; D000098662:Vasodilator-Stimulated Phosphoprotein; D019000:Phosphotyrosine; D019877:Hydroxymethyl and Formyl Transferases; C107023:p80(NPM-ALK) protein; D011505:Protein-Tyrosine Kinases; D009709:Nucleotide Deaminases; D008727:Methotrexate