Abstract
In search for general PCR targets for minimal residual disease (MRD) studies in acute myeloid leukemia (AML), Wilms' tumor gene 1 (WT1) expression was assessed by real-time RT-PCR relative to the control gene ABL in 569 archived samples of AML patients (pts). Pts were analyzed at diagnosis (n=116) and during follow-up (n=105, median 4 times, range 2–17). Median follow-up time was 258 days (range 16–1578 days). In 66 pts, the WT1 expression was analyzed in comparison to a second PCR marker or to multiparameter flow cytometry. Quantitative WT1 levels correlated to the clinical course or a second marker in 83–96% of the cases. Prognostic significance of WT1 levels was analyzed at diagnosis and three intervals: (1) days 16–60, (2) days 61–120, and (3) days 121–180 after start of chemotherapy. Higher levels of WT1 expression were associated with shorter overall survival (OS) and event-free survival (EFS) within intervals 2 and 3 but not at diagnosis or interval 1. In addition, within these intervals, WT1/ABL levels ⩽0.4% were associated with improved OS and EFS. An increase of WT1 levels was detected in 16/44 cases, which subsequently relapsed within a median of 38 days (range 8–180 days). In conclusion, quantification of WT1 may be used for MRD studies and for prognostification in AML.
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References
Schnittger S, Weisser M, Schoch C, Hiddemann W, Haferlach T, Kern W . New score predicting for prognosis in PML-RARA+, AML1-ETO+, or CBFBMYH11+ acute myeloid leukemia based on quantification of fusion transcripts. Blood 2003; 102: 2746–2755.
Buonamici S, Ottaviani E, Testoni N, Montefusco V, Visani G, Bonifazi F et al. Real-time quantitation of minimal residual disease in inv(16)-positive acute myeloid leukemia may indicate risk for clinical relapse and may identify patients in a curable state. Blood 2002; 99: 443–449.
Guerrasio A, Pilatrino C, De Micheli D, Cilloni D, Serra A, Gottardi E et al. Assessment of minimal residual disease (MRD) in CBFbeta/MYH11-positive acute myeloid leukemias by qualitative and quantitative RT-PCR amplification of fusion transcripts. Leukemia 2002; 16: 1176–1181.
Cilloni D, Saglio G . WT1 as a universal marker for minimal residual disease detection and quantification in myeloid leukemias and in myelodysplastic syndrome. Acta Haematol 2004; 112: 79–84.
Call KM, Glaser T, Ito CY, Buckler A, Pelletier J, Haber DA et al. Isolation and characterization of a zinc finger polypeptide gene at the human chromosome 11 Wilms' tumor locus. Cell 1990; 60: 509–520.
Cook DM, Hinkes MT, Bernfield M, Rauscher III FJ . Transcriptional activation of the syndecan-1 promoter by the Wilms' tumor protein WT1. Oncogene 1996; 13: 1789–1799.
Englert C, Hou X, Maheswaran S, Bennett P, Ngwu C, Re GG et al. WT1 suppress synthesis of the epidermal growth factor receptor and induces apoptosis. EMBO J 1995; 14: 4662–4675.
Miwa H, Beran M, Saunders GF . Expression of the Wilms' tumor gene (WT1) in human leukemias. Leukemia 1992; 6: 405–409.
Miyagi T, Ahuja H, Kubonishi I, Koeffler HP, Miyoshi I . Expression of the candidate Wilm's tumor gene, WT1, in human leukemia cells. Leukemia 1993; 7: 970–977.
Inoue K, Ogawa H, Sonoda Y, Kimura T, Sakabe H, Oka Y et al. Aberrant overexpression of the Wilms tumor gene (WT1) in human leukemia. Blood 1997; 89: 1405–1412.
Menssen HD, Renkl HJ, Rodeck U, Maurer J, Notter M, Schwaz S et al. Presence of Wilms tumor gene (WT1) transcripts and the WT1 nuclear protein in the majority of human acute leukemias. Leukemia 1995; 9: 1060–1067.
Algar EM, Khromykh T, Smith SI, Blackburn DM, Bryson GJ, Smith PJ . A WT1 antisense oligonucleotide inhibits proliferation and induces apoptosis in myeloid leukemia cell lines. Oncogene 1996; 12: 1005–1014.
Deuel TF, Guan LS, Wang ZY . Wilms' tumor gene product WT1 arrests macrophage differentiation of HL60 cells through its zinc-finger domain. Biochem Biophys Res Commun 1999; 254: 192–196.
Smith SI, Weil D, Johnson GR, Boyd AW, Li CL . Expression of the Wilms' tumor suppressor gene, WT1 is upregulated by leukemia inhibitory factor and induces monocytic differentiation in M1 leukemic cells. Blood 1998; 91: 764–773.
Svedberg H, Chylicki K, Baldetorp B, Rauscher III FL, Gullberg U . Constitutive expression of the Wilms' tumor gene (WT1) in the leukemic cell line U937 blocks part of the differentiation program. Oncogene 1998; 16: 925–932.
Maurer U, Brieger J, Weidmann E, Mitrou PS, Hoelzer D, Bergmann L . The Wilms' tumor gene is expressed in a subset of CD34+ progenitors and downregulated early in the course of differentiation in vitro. Exp Hematol 1997; 25: 945–950.
Menssen HD, Renkl HJ, Entezami M, Thiel E . Wilms' tumor gene expression in human CD34+ hematopoietic progenitors during fetal development and early clonogenic growth. Blood 1997; 89: 3486–3487.
Sekiya M, Adachi M, Hinoda Y, Imai K, Yachi A . Downregulation of the Wilms' tumor gene (wt1) during myelomonocytic differentiation in HL60 cells. Blood 1994; 83: 1876–1882.
Cilloni D, Gottardi E, De Micheli D, Serra A, Volpe G, Messa F et al. Quantitative assessment of WT1 expression by real time quantitative PCR may be a useful tool for monitoring minimal residual disease in acute leukemia patients. Leukemia 2002; 16: 2115–2121.
Trka J, Kalinova M, Hrusak O, Zuna J, Krejci O, Madzo J et al. Real time quantitative PCR detection of WT1 gene expression in children with AML: prognostic significance, correlation with disease status and residual disease detection by flow cytometry. Leukemia 2002; 16: 1381–1389.
Ostergaard M, Olesen LH, Hasle H, Kjeldsen E, Hokland P . WT1 gene expression: an excellent tool for monitoring minimal residual disease in 70% of acute myeloid leukaemia patients – results from a single-centre study. Br J Haematol 2004; 125: 590–600.
Ogawa H, Tamaki H, Ikegame K, Soma T, Kawakami M, Tsuboi A et al. The usefulness of monitoring WT1 gene transcripts for the prediction and management of relapse following allogeneic stem cell transplantation in acute type leukemia. Blood 2003; 101: 1698–1704.
Inoue K, Ogawa H, Yamagami T, Soma T, Tani Y, Tatekawa T et al. Long-term follow-up of minimal residual disease in leukemia patients by monitoring WT1 (Wilms tumor gene) expression levels. Blood 1996; 88: 2267–2278.
Bergmann L, Maurer U, Weidmann E . Wilms tumor gene expression in acute myeloid leukemias. Leuk Lymphoma 1997; 25: 435–443.
Garg M, Moore H, Tobal K, Liu Yin JA . Prognostic significance of quantitative analysis of WT1 gene transcripts by competitive reverse transcription polymerase chain reaction in acute leukaemia. Br J Haematol 2003; 123: 49–59.
Gaiger A, Schmid D, Heinze G, Linnerth B, Greinix H, Halhs P et al. Detection of the WT1 transcript by RT-PCR in complete remission has no prognostic relevance in de novo acute myeloid leukemia. Leukemia 1998; 12: 1886–1894.
Schmid D, Heinze G, Linnerth B . Prognostic significance of WT1 gene expression at diagnosis in adult de novo acute myeloid leukemia. Leukemia 1997; 11: 639–643.
Inoue K, Sugiyama H, Ogawa, Nakagawa M, Yamagami T, Miwa H et al. WT1 as a new prognostic factor and a new marker for the detection of minimal residual disease in acute leukemia. Blood 1994; 84: 3071–3079.
Bergmann L, Mieting C, Maurer U, Brieger J, Karakas T, Weidmann E et al. High levels of Wilms' tumor gene (wt1) mRNA in acute myeloid leukemias are associated with a worse long term outcome. Blood 1997; 90: 1217–1225.
Bennett JM, Catovsky D, Daniel MT, Flandrin G, Galton DA, Gralnick HR et al. Proposals for the classification of the acute leukaemias. French–American–British (FAB) co-operative group. Br J Haematol 1976; 33: 451–458.
Bennett JM, Catovsky D, Daniel MT, Flandrin G, Galton DA, Gralnick HR et al. Proposal for the recognition of minimally differentiated acute myeloid leukaemia (AML-MO). Br J Haematol 1991; 78: 325–329.
Jaffe ES, Harris NL, Stein H, Vardiman JW . World Health Organization Classification of Tumours. Pathology and Genetics of Tumours of Haematopoietic and Lymphoid Tissues. Lyon, France: IARC Press, 2001.
Buchner T, Hiddemann W, Wormann B, Loffler H, Gassmann W, Haferlach T et al. Double induction strategy for acute myeloid leukemia: the effect of high-dose cytarabine with mitoxantrone instead of standard-dose cytarabine with daunorubicin and 6-thioguanine: a randomized trial by the German AML Cooperative Group. Blood 1999; 93: 4116–4124.
Buchner T, Hiddemann W, Berdel WE, Wormann B, Schoch C, Fonatsch C, et al, German AML Cooperative Group. 6-Thioguanine, cytarabine, and daunorubicin (TAD) and high-dose cytarabine and mitoxantrone (HAM) for induction, TAD for consolidation, and either prolonged maintenance by reduced monthly TAD or TAD–HAM–TAD and one course of intensive consolidation by sequential HAM in adult patients at all ages with de novo acute myeloid leukemia (AML): a randomized trial of the German AML Cooperative Group. J Clin Oncol 2003; 21: 4496–5004.
Gabert J, Beillard E, van der Velden VH, Bi W, Grimwade D, Pallisgaard N et al. Standardization and quality control studies of ‘real-time’ quantitative reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia – a Europe Against Cancer program. Leukemia 2003; 17: 2318–2357.
Schnittger S, Schoch C, Kern W, Hiddemann W, Haferlach T . FLT3 length mutations as marker for follow-up studies in acute myeloid leukaemia. Acta Haematol 2004; 112: 68–78.
Kern W, Voskova D, Schoch C, Hiddemann W, Schnittger S, Haferlach T . Determination of relapse risk based on assessment of minimal residual disease during complete remission by multiparameter flow cytometry in unselected patients with acute myeloid leukemia. Blood 2004; 104: 3078–3085.
Kreuzer KA, Saborowski A, Lupberger J, Appelt C, Na IK, Le Coutre P et al. Fluorescent 5′-exonuclease assay for the absolute quantification of Wilms' tumour gene (WT1) mRNA: implications for monitoring human leukemias. Br J Haematol 2001; 114: 313–318.
Elmaagacli AH, Beelen DW, Trenschel R, Schaefer UW . The detection of WT1 transcripts is not associated with an increased leukemic relapse rate in patients with acute leukemia after allogeneic bone marrow or peripheral blood stem cell transplantation. Bone Marrow Transplant 2000; 25: 91–96.
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Weisser, M., Kern, W., Rauhut, S. et al. Prognostic impact of RT-PCR-based quantification of WT1 gene expression during MRD monitoring of acute myeloid leukemia. Leukemia 19, 1416–1423 (2005). https://doi.org/10.1038/sj.leu.2403809
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DOI: https://doi.org/10.1038/sj.leu.2403809
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