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

A. M. Willasch, B. Gruhn, T. Coliva, M. Kalinova, G. Schneider, H. Kreyenberg, D. Steinbach, G. Weber, I. H.I.M. Hollink, C. M. Zwaan, A. Biondi, V. H.J. van der Velden, D. Reinhardt, G. Cazzaniga, P. Bader, J. Trka

Research output: Contribution to journalArticlepeer-review

50 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1472-1479
Number of pages8
JournalLeukemia
Volume23
Issue number8
DOIs
Publication statusPublished - 2009
Externally publishedYes

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