A standardized, sensitive and universal method for minimal residual disease (MRD) detection in acute myeloid leukemia (AML) is still pending. Although hyperexpression of Wilms' tumor (WT1) gene transcript has been frequently proposed as an MRD marker in AML, wide comparability of the various methods used for evaluating WT1 expression has not been given. We established and standardized a multicenter approach for quantifying WT1 expression by quantitative reverse transcriptase PCR (qRT-PCR), on the basis of a primer/probe set combination at exons 6 and 7. In a series of quality-control rounds, we analyzed 69 childhood AML samples and 47 normal bone marrow (BM) samples from 4 participating centers. Differences in the individual WT1 expressions levels ranged within <0.5 log of the mean in 82% of the cases. In AML samples, the median WT1/1E+04 Abelson (ABL) expression was 3.5E+03 compared with that of 2.3E+01 in healthy BM samples. As 11.5% of childhood AML samples in this cohort harbored WT1 mutations in exon 7, the effect of mutations on WT1 expression has been investigated, showing that mutated cases expressed significantly higher WT1 levels than wild-type cases. Hence, our approach showed high reproducibility and applicability, even in patients with WT1 mutations; therefore, it can be widely used for the quantitation of WT1 expression in future clinical trials.

Willasch, A., Gruhn, B., Coliva, T., Kalinova, M., Schneider, G., Kreyenberg, H., et al. (2009). Standardization of WT1 mRNA quantitation for minimal residual disease monitoring in childhood AML and implications of WT1 gene mutations: a European multicenter study. LEUKEMIA, 23(8), 1472-1479 [10.1038/leu.2009.51].

Standardization of WT1 mRNA quantitation for minimal residual disease monitoring in childhood AML and implications of WT1 gene mutations: a European multicenter study

COLIVA, TIZIANA ANGELA;BIONDI, ANDREA;Cazzaniga, G;
2009

Abstract

A standardized, sensitive and universal method for minimal residual disease (MRD) detection in acute myeloid leukemia (AML) is still pending. Although hyperexpression of Wilms' tumor (WT1) gene transcript has been frequently proposed as an MRD marker in AML, wide comparability of the various methods used for evaluating WT1 expression has not been given. We established and standardized a multicenter approach for quantifying WT1 expression by quantitative reverse transcriptase PCR (qRT-PCR), on the basis of a primer/probe set combination at exons 6 and 7. In a series of quality-control rounds, we analyzed 69 childhood AML samples and 47 normal bone marrow (BM) samples from 4 participating centers. Differences in the individual WT1 expressions levels ranged within <0.5 log of the mean in 82% of the cases. In AML samples, the median WT1/1E+04 Abelson (ABL) expression was 3.5E+03 compared with that of 2.3E+01 in healthy BM samples. As 11.5% of childhood AML samples in this cohort harbored WT1 mutations in exon 7, the effect of mutations on WT1 expression has been investigated, showing that mutated cases expressed significantly higher WT1 levels than wild-type cases. Hence, our approach showed high reproducibility and applicability, even in patients with WT1 mutations; therefore, it can be widely used for the quantitation of WT1 expression in future clinical trials.
Articolo in rivista - Articolo scientifico
Sensitivity and Specificity; Adolescent; Middle Aged; Neoplasm Proteins; Bone Marrow Examination; Neoplasm, Residual; RNA, Neoplasm; Cohort Studies; Leukemia, Myeloid; DNA Primers; Child; Adult; WT1 Proteins; Male; Young Adult; Infant; Female; Child, Preschool; Gene Expression Regulation, Leukemic; Humans; Genes, Wilms Tumor; Acute Disease; RNA, Messenger; Reverse Transcriptase Polymerase Chain Reaction; Exons
English
2009
23
8
1472
1479
none
Willasch, A., Gruhn, B., Coliva, T., Kalinova, M., Schneider, G., Kreyenberg, H., et al. (2009). Standardization of WT1 mRNA quantitation for minimal residual disease monitoring in childhood AML and implications of WT1 gene mutations: a European multicenter study. LEUKEMIA, 23(8), 1472-1479 [10.1038/leu.2009.51].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/14933
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