title: Multiplex real-time RT-PCR for prospective evaluation of WT1 and fusion gene transcripts in newly diagnosed de novo acute myeloid leukemia.
abstract: Prognostic assessment is crucial for the management of AML. Although the use of karyotype analysis for risk-stratification is widely accepted, prognosis of AML remains ambiguous, particularly for patients categorized into the intermediate cytogenetic risk group and additional markers are required for an accurate prediction of outcome. For this study, we used multiplex real-time RT-PCR, which can simultaneously quantify WT1 and 10 distinct fusion gene transcripts, to prospectively evaluate the pre-treatment bone marrow findings of 53 de novo AML patients. Five patients with normal karyotype or insufficient metaphases detected by conventional karyotype analysis proved to have AML1-MTG8, CBFbeta-MYH11 or PML-RARalpha fusion transcripts. WT1 overexpression was observed in 92% of the patients, and the levels were significantly higher in the cytogenetic favorable risk group, especially patients with PML-RARalpha. WT1 levels also correlated with the percentage of blasts in bone marrow, especially in cases of core-binding factor leukemia. There was no association between initial WT1 levels and outcome in terms of event-free survival or overall survival. These results suggest that multiplex real-time RT-PCR is rapid and useful for the precise cytogenetic stratification of AML, and that WT1 levels at presentation correlate with several biologic features of leukemia, but have no prognostic significance.
authors.affiliation: Department of Hematology, Nagoya University Graduate School of Medicine, Nagoya, Japan. myanada@med.nagoya-u.ac.jp
authors.forename: Masamitsu
authors.initials: M
authors.lastname: Yanada
authors.forename: Seitaro
authors.initials: S
authors.lastname: Terakura
authors.forename: Toshiya
authors.initials: T
authors.lastname: Yokozawa
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Kiyoi
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Emi
authors.forename: Kunio
authors.initials: K
authors.lastname: Kitamura
authors.forename: Akio
authors.initials: A
authors.lastname: Kohno
authors.forename: Masafumi
authors.initials: M
authors.lastname: Tanaka
authors.forename: Tadasu
authors.initials: T
authors.lastname: Tobita
authors.forename: Takaaki
authors.initials: T
authors.lastname: Takeo
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sao
authors.forename: Takae
authors.initials: T
authors.lastname: Kataoka
authors.forename: Masahide
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Akihiro
authors.initials: A
authors.lastname: Takeshita
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Morishita
authors.forename: Tomoki
authors.initials: T
authors.lastname: Naoe
authors.forename: Isamu
authors.initials: I
authors.lastname: Sugiura
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001853:Bone Marrow / Q000378:metabolism; D005260:Female; D006801:Humans; D007948:Leukemia, Monocytic, Acute / Q000175:diagnosis* / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D011446:Prospective Studies; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015996:Survival Rate; D014158:Transcription, Genetic / Q000235:genetics*; D016896:Treatment Outcome; D025721:WT1 Proteins / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D012334:RNA, Neoplasm; D011993:Recombinant Fusion Proteins; D025721:WT1 Proteins