D. Hoelzer

Thymic adult T-cell acute lymphoblastic leukemia stratified in standard- and high-risk group by aberrant HOX11L2 expression: experience of the German multicenter ALL study group

Adult T-cell acute lymphoblastic leukemia (T-ALL) continues to represent an unfavorable disease. Molecularly based treatment stratifications could help improve outcome. The prognostic impact of HOX11 and HOX11L2 expression has been an area of controversy. We have investigated 286 adult T-ALL patients enrolled into the German Multicenter ALL (GMALL) therapy protocols by comparative real-time RT-PCR. High HOX11 expression and HOX11L2 expression were predominantly seen in thymic T-ALL (P⩽0.031). In a multivariate analysis HOX11L2 expression proved to be an independent adverse risk factor for relapse-free survival (RFS) with a hazard ratio (HR) of 2.02 (P=0.023) and an HR for overall survival (OS) of 1.81 (P=0.021), both adjusted for the immunophenotype. HOX11 expression was found to have a favorable impact on RFS (HR 0.51; P=0.048) but did not exhibit a significant impact on OS. A subgroup analysis for thymic T-ALL revealed a more pronounced negative correlation of HOX11L2 expression with RFS (HR 3.26; P=0.002) and OS (HR 2.38; P=0.009). Although the prognostic impact of HOX11 in T-ALL is less clear, HOX11L2 expression identifies a small subset of high-risk patients, who are so far classified as standard-risk group. Thus, patients with aberrant HOX11L2 expression should be considered early as candidates for intensified treatment regimes.

High rate of chromosome abnormalities detected by fluorescence in situ hybridization using BCR and ABL probes in adult acute lymphoblastic leukemia

The value of dual-color fluorescence in situ hybridization (FISH) with BCR and ABL probes for the detection of the Philadelphia (Ph) translocation and of other alterations involving ABL and/or BCR was evaluated in adult acute lymphoblastic leukemia (ALL). One hundred and four patients were studied prospectively using interphase nuclei FISH, chromosome analysis (CA), and PCR assays for the chimeric BRC/ABL transcript. FISH detected a Ph translocation in 24 cases (23.1%), as was confirmed by CA and/or PCR. FISH revealed a false positive diagnosis of a Ph translocation in four cases (5% false positive rate). Among 54 cases with combined FISH, CA and PCR assays, FISH failed to establish a correct diagnosis in 3.7%, PCR in 5.6%, and CA in 7.4%. The combination of two screening methods led to discrepant results in 9.3% (FISH + PCR), 11.1% (FISH + CA), or 13% (CA + PCR) of the cases. In seven of 80 (8.8%) Ph-negative patients, gain of BCR and/or ABL was identified. Overall, FISH detected alterations of the BCR and/or ABL genes with an incidence of 29.8% of the current study. Due to the possibility of false positive diagnosis of a Ph translocation using dual-color FISH the combination with chromosome and/or RT-PCR analyses is recommended in adult ALL patients.

The homeobox gene CDX2 is aberrantly expressed and associated with an inferior prognosis in patients with acute lymphoblastic leukemia.

Molecular characterization of acute lymphoblastic leukemia (ALL) has greatly improved the ability to categorize and prognostify patients with this disease. In this study, we show that the proto-oncogene CDX2 is aberrantly expressed in the majority of cases with B-lineage ALL and T-ALL. High expression of CDX2 correlated significantly with the ALL subtype pro-B ALL, cALL, Ph+ ALL and early T-ALL. Furthermore, high expression of CDX2 was associated with inferior overall survival and showed up as a novel and strong risk factor for ALL in bivariate analysis. Functional analyses showed that overexpression of Cdx2 in murine bone marrow progenitors perturbed genes involved in lymphoid development and that depletion of CDX2 in the human ALL cell line Nalm6 inhibited colony formation. These data indicate that aberrant CDX2 expression occurs frequently and has prognostic impact in adult patients with ALL.

Prospective BCR-ABL analysis by polymerase chain reaction (RT-PCR) in adult acute B-lineage lymphoblastic leukemia: reliability of RT-nested-PCR and comparison to cytogenetic data.

The reliability of routine BCR-ABL RT-nested-PCR was evaluated in 1453 B-lineage ALL or hybrid leukemia at initial diagnosis by RT-nested-PCR. All BCR-ABL-positive (n = 642) and 176 BCR-ABL-negative samples underwent a second RT-PCR. In 518 patients, karyotyping and/or FISH was compared to the BCR-ABL status. The second RT-PCR revealed in 155/642 initially positive samples a divergent result (153 BCR-ABL-negative, two other transcripts) that in most cases turned out to be caused by contaminations in the first RT-nested-PCR. Confirmatory RT-PCR detected 2/176 false negative first RT-nested-PCR results. Thirty-nine specimens remained ambiguous despite different RT-PCR approaches. As far as cytogenetic evaluation and FISH is available (n = 23), the majority but not all patients with an ambiguous RT-PCR result were Ph-negative (n = 18). RT-nested-PCR and cytogenetics yielded in 346 of 383 evaluable samples a concordant result. Differing results are given and account in part to the lower sensitivity of karyotyping. Taken together, confirmed RT-PCR detected BCR-ABL fusion transcripts consistently in 487 out of 1453 ALL samples (c-ALL: 43%, pre-B ALL: 34%, pro-B ALL: 5%, B-ALL: 0%, hybrid leukemia: 5/11). Since false positive initial RT-nested-PCR data were frequent, either confirmatory second RT-PCR or FISH analysis is warranted to guarantee sensitive and reliable results of utmost clinical relevance.

The German competence network 'Acute and chronic leukemias'

R Hehlmann, U Berger, C Aul, Th Büchner, H Döhner, G Ehninger, A Ganser, N Gökbuget, D Hoelzer, K Überla, W Gassmann, WD Ludwig, H Rieder, M Kneba, A Hochhaus, A Reiter, W Hiddemann, OG Ottmann, U Germing, K Adelhard, M Dugas, P Dirschedl, D Messerer, A Böhme, E Harrison-Neu, M Griesshammer, J Kienast, HJ Kolb, AD Ho, M Hallek, A Neubauer, B Schlegelberger, D Niederwieser, G Heil, T Müller and J Hasford Kompetenznetz ‘Akute und chronische Leukämien’, III. Medizinische Universitätsklinik, Klinikum Mannheim der Universität Heidelberg, Mannheim, Germany

Uncovering early, lineage-dependent effects of TPMT genotype in adult acute lymphoblastic leukemia by minimal residual disease

Uncovering early, lineage-dependent effects of TPMT genotype in adult acute lymphoblastic leukemia by minimal residual disease

Treatment of Minimal Residual Disease in Adult ALL: The German National Study

Results in adult acute lymphoblastic leukemia (ALL) have clearly improved during the last decade with intensified chemotherapy. Complete re-mission (CR) rates of 70% – 85% and 5-year disease-free survival rates of 30% – 40% or even more can now be achieved (1 – 6). The better out-come in adult ALL is due to experience gained in childhood ALL, such as the selection and scheduling of cytostatic drugs, the use of early intensive consolidation therapy and the evaluation of various modalities for prophylactic treatment of the nervous system (CNS).

Prof. Thomas Buchner (22 September 1934-5 August 2016) OBITUARY

Prof. Thomas Buchner (22 September 1934–5 August 2016)—A leading European hematologist and AML trialist

Prof. Thomas Büchner (22 September 1934–5 August 2016)—A leading European hematologist and AML trialist

Prof. Thomas Buchner (22 September 1934–5 August 2016)—A leading European hematologist and AML trialist

LeukemiaNet: A transnational model for cooperative leukemia research.

6556 Background: Leukemia research may be jeopardized by lack of communication structures or fragmentation of study activities, insufficient interaction and underestimation of the problem. METHODS Recognizing that the only way to cure leukemia is by cooperative research, scientists in Europe in 2002, set out to establish an exemplary cooperative network for research in leukemia, the European LeukemiaNet (ELN). At present, ELN integrates 105 national leukemia trial groups and networks, 105 interdisciplinary specialty groups and about 1000 leukemia specialists from 175 institutions. They care for ten thousands of leukemia patients in 33 countries. RESULTS The most visible results are 1.) those due to the cooperative research projects and trials as reflected by a large number of high impact publications; and 2.) the guidelines and management recommendations for virtually every type of leukemia and interdisciplinary specialty which have lain the groundwork for uniform definitions and standards required for common clinical trials and projects. CONCLUSIONS ELN is a model of transnational cooperation. Working together successfully has created a spirit of cooperation and mutual trust. Due to its structured and long-term cooperation, ELN is likely to have a durable impact on leukemia research in Europe. By promoting cooperation over competition that is necessary for good research, ELN provides a competitive advantage for all participants to the benefit of every patient with leukemia worldwide.