title: Presence of the BCR-ABL mutation Glu255Lys prior to STI571 (imatinib) treatment in patients with Ph+ acute lymphoblastic leukemia.
abstract: The tyrosine kinase inhibitor STI571 (imatinib) binds competitively to the adenosine triphosphate (ATP) binding site of the ABL kinase, thereby inhibiting auto- and substrate phosphorylation of the oncogenic protein BCR-ABL and preventing the activation of downstream signaling pathways. Comparative studies on leukemic cell samples obtained from chronic myelogenous leukemia (CML) and Philadelphia chromosome-positive (Ph+) acute lymphoblastic leukemia (ALL) patients before and after treatment with STI571 reported point mutations in resistant samples after a short time of therapy. The aim of this study was to determine whether patients with Ph+ ALL in whom resistance developed as a consequence of the Glu255Lys mutation already harbored this subclone prior to STI571 treatment. First, the migration pattern of cDNAs from 30 bone marrow samples from patients with Ph+ ALL was analyzed by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). Thereafter, detailed mutational analysis using genomic DNA was performed on initial STI571-naive bone marrow samples of 4 individuals with Ph+ ALL, for whom the mutation Glu255Lys in association with STI571 treatment had been shown. A 166-bp PCR fragment spanning from nucleotide (nt) 862 to nt 1027 was cloned, and 108 clones per sample were analyzed by direct sequencing. This more sensitive technique revealed the presence of the Glu255Lys mutation in 2 initial samples, one clone each. We identified for the first time the mutation Glu255Lys in STI571-naive leukemic samples of Ph+ ALL patients. The findings suggest that the mutation exists in a very small subpopulation of leukemic cells at the beginning of STI571 therapy.
authors.affiliation: Department of Hematology, University Hospital, Theodor-Stern-Kai 7, 60596, Frankfurt/Main, Germany. w.k.hofmann@em.uni-frankfurt.de
authors.forename: Wolf-Karsten
authors.initials: WK
authors.lastname: Hofmann
authors.forename: Martina
authors.initials: M
authors.lastname: Komor
authors.forename: Barbara
authors.initials: B
authors.lastname: Wassmann
authors.forename: Letetia C
authors.initials: LC
authors.lastname: Jones
authors.forename: Harald
authors.initials: H
authors.lastname: Gschaidmeier
authors.forename: Dieter
authors.initials: D
authors.lastname: Hoelzer
authors.forename: H Phillip
authors.initials: HP
authors.lastname: Koeffler
authors.forename: Oliver G
authors.initials: OG
authors.lastname: Ottmann
mesh_terms: D000328:Adult; D019943:Amino Acid Substitution; D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D001549:Benzamides; D001853:Bone Marrow / Q000201:enzymology; D017322:Clinical Trials, Phase II as Topic; D002999:Clone Cells / Q000201:enzymology; D015331:Cohort Studies; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000494:pharmacology / Q000627:therapeutic use*; D016044:Fusion Proteins, bcr-abl / Q000037:antagonists & inhibitors / Q000235:genetics*; D006801:Humans; D000068877:Imatinib Mesylate; D020125:Mutation, Missense; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D014411:Neoplastic Stem Cells / Q000201:enzymology*; D010879:Piperazines / Q000494:pharmacology / Q000627:therapeutic use*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy / Q000201:enzymology / Q000235:genetics*; D011743:Pyrimidines / Q000494:pharmacology / Q000627:therapeutic use*; D012641:Selection, Genetic; D013997:Time Factors
chemical_list: D000970:Antineoplastic Agents; D001549:Benzamides; D018076:DNA, Complementary; D004273:DNA, Neoplasm; D004791:Enzyme Inhibitors; D009363:Neoplasm Proteins; D010879:Piperazines; D011743:Pyrimidines; D000068877:Imatinib Mesylate; D016044:Fusion Proteins, bcr-abl