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[show abstract]
[hide abstract]
ABSTRACT: Minimal residual disease (MRD) diagnostics is of high clinical relevance in patients with indolent B-cell Non-Hodgkin lymphomas (B-NHL) and serves as a surrogate parameter to evaluate treatment effectiveness and long-term prognosis. MRD diagnostics performed by real-time quantitative PCR (RQ-PCR) is the gold-standard and currently the most sensitive and the most broadly applied method in follicular lymphoma (FL) and mantle cell lymphoma (MCL). RQ-PCR analysis of the junctional regions of the rearranged immunoglobulin heavy-chain gene (IgH) serves as the most broadly applicable MRD target in B-NHL (∼80%). Chromosomal translocations as t(14;18) translocation in FL and t(11;14) translocation in MCL can be used in selected lymphoma subtypes. In patients with B-cell chronic lymphocytic leukemia, both flow-cytometry as well as RQ-PCR are equally suitable for MRD assessment as long as a sensitivity of ≤10(-4) shall be achieved.MRD diagnostics targeting the IgH gene is complex and requires extensive knowledge and experience because the junctional regions of each lymphoma have to be identified before the patient-specific RQ-PCR assays can be designed for MRD monitoring. Furthermore, somatic mutations of the IgH region occurring during B-cell development of germinal center and post-germinal center lymphomas may hamper appropriate primer binding leading to false negative results. The translocations mentioned above have the advantage that consensus forward primers and probes, both placed in the breakpoint regions of chromosome 18 in FL and chromosome 11 in MCL, can be used in combination with a reverse primer placed in the IgH joining region of chromosome 14. RQ-PCR-based methods can reach a good sensitivity (≤10(-4)). This chapter provides all relevant background information and technical aspects for the complete laboratory process from detection of the clonal IgH gene rearrangement and the chromosomal translocations at diagnosis to the actual MRD measurements in clinical follow-up samples of B-NHL. However, it should be noted that MRD diagnostics for clinical treatment protocols has to be accompanied by regular international quality control rounds to ensure the reproducibility and reliability of the MRD results.
Methods in molecular biology (Clifton, N.J.) 01/2013; 971:175-200.
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Julia Richter,
Matthias Schlesner,
Steve Hoffmann,
Markus Kreuz,
Ellen Leich,
Birgit Burkhardt,
Maciej Rosolowski,
Ole Ammerpohl,
Rabea Wagener,
Stephan H Bernhart, [......],
Peter F Stadler,
Peter Lichter,
Roland Eils,
Ralf Küppers,
Michael Hummel,
Wolfram Klapper,
Philip Rosenstiel,
Andreas Rosenwald,
Benedikt Brors,
Reiner Siebert
[show abstract]
[hide abstract]
ABSTRACT: Burkitt lymphoma is a mature aggressive B-cell lymphoma derived from germinal center B cells. Its cytogenetic hallmark is the Burkitt translocation t(8;14)(q24;q32) and its variants, which juxtapose the MYC oncogene with one of the three immunoglobulin loci. Consequently, MYC is deregulated, resulting in massive perturbation of gene expression. Nevertheless, MYC deregulation alone seems not to be sufficient to drive Burkitt lymphomagenesis. By whole-genome, whole-exome and transcriptome sequencing of four prototypical Burkitt lymphomas with immunoglobulin gene (IG)-MYC translocation, we identified seven recurrently mutated genes. One of these genes, ID3, mapped to a region of focal homozygous loss in Burkitt lymphoma. In an extended cohort, 36 of 53 molecularly defined Burkitt lymphomas (68%) carried potentially damaging mutations of ID3. These were strongly enriched at somatic hypermutation motifs. Only 6 of 47 other B-cell lymphomas with the IG-MYC translocation (13%) carried ID3 mutations. These findings suggest that cooperation between ID3 inactivation and IG-MYC translocation is a hallmark of Burkitt lymphomagenesis.
Nature Genetics 11/2012; · 35.53 Impact Factor
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Martin Dreyling,
Hanneke C Kluin-Nelemans,
Sílvia Beà,
Wolfram Klapper,
Niclas Vogt,
Marie-Helene Delfau-Larue,
Grit Hutter,
Chan Cheah,
Annalisa Chiappella,
Sergio Cortelazzo, Christiane Pott,
Georg Hess,
Carlo Visco,
Umberto Vitolo,
Pawel Klener,
Igor Aurer,
Michael Unterhalt,
Vincent Ribrag,
Eva Hoster,
Olivier Hermine
[show abstract]
[hide abstract]
ABSTRACT: Abstract Mantle cell lymphoma (MCL) is a distinct subtype of malignant lymphoma characterized by the chromosomal translocation t(11;14)(q13;q32) resulting in constitutional overexpression of cyclin D1 and cell cycle dysregulation in virtually all cases. Clinically, MCL displays an aggressive clinical course with a continuous relapse pattern and a median survival of only 3-7 years. However, a subset of up to 15% long-term survivors has recently been identified with a rather indolent clinical course. In general, conventional chemotherapy is only palliative and median duration of remissions is only 1-2 years. In 2000, the European MCL Network ( http://www.european-mcl.net ) was founded, which consists of 15 national lymphoma study groups supplemented by experts in hematopathology, cytogenetics and molecular genetics. During the last decade, the European consortium has successfully initiated the largest phase III trials in MCL worldwide. In the current study generation, the addition of high dose Ara-C to an R-CHOP like regimen followed by myeloablative consolidation achieved a significant improvement of progression-free survival. Similarly, in elderly patients rituximab maintenance until progression led to a marked prolongation of remission duration. Emerging strategies include proteasome inhibitors, immune modulatory drugs (IMiDs), mTOR inhibitors and others, all based on the dysregulated cell cycle machinery and impairment of several signaling transduction and apoptotic pathways. Future strategies will apply individualized approaches according to the molecular risk profile of the patient. At the annual conference in Lisbon, recent results of molecular pathogenesis, analyses of current clinical trials and new study concepts were discussed.
Leukemia & lymphoma 09/2012; · 2.40 Impact Factor
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Alba Navarro,
Guillem Clot,
Cristina Royo,
Pedro Jares,
Anastasia Hadzidimitriou,
Andreas Agathangelidis,
Vasilis Bikos,
Nikos Darzentas,
Theodora Papadaki,
Itziar Salaverria, [......],
Birgitta Sander,
German Ott,
Andreas Rosenwald,
Dolors Colomer,
Eva Giné,
Reiner Siebert,
Armando Lopez-Guillermo,
Kostas Stamatopoulos,
Sílvia Beà,
Elías Campo
[show abstract]
[hide abstract]
ABSTRACT: Mantle cell lymphoma (MCL) is a heterogeneous disease with most patients following an aggressive clinical course, whereas others having an indolent behavior. We conducted an integrative and multidisciplinary analysis of 177 MCL to determine whether the immunogenetic features of the clonotypic B-cell receptors (BcR) may identify different subsets of tumors. Truly unmutated (100% identity) IGHV genes were found in 24% cases, 40% were minimally/borderline mutated (99.9%-97%), 19% significantly mutated (96.9%-95%), and 17% hypermutated (<95%). Tumors with high or low mutational load used different IGHV genes, and their gene expression profiles were also different for several gene pathways. A gene set enrichment analysis showed that MCL with high and low IGHV mutations were enriched in memory and naive B-cell signatures, respectively. Furthermore, the highly mutated tumors had less genomic complexity, were preferentially SOX11-negative, and showed more frequent nonnodal disease. The best cut-off of germline identity of IGHV genes to predict survival was 97%. Patients with high and low mutational load had significant different outcome with 5-year overall survival (OS) of 59% and 40%, respectively (P = 0.004). Nodal presentation and SOX11 expression also predicted for poor OS. In a multivariate analysis, IGHV gene status and SOX11 expression were independent risk factors. In conclusion, these observations suggest the idea that MCL with mutated IGHV, SOX11-negativity, and nonnodal presentation correspond to a subtype of the disease with more indolent behavior. Cancer Res; 72(20); 5307-16. ©2012 AACR.
Cancer Research 08/2012; 72(20):5307-5316. · 7.86 Impact Factor
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Heiner Zimmermann,
Ilske Oschlies,
Susanne Fink, Christiane Pott,
Hans H Neumayer,
Hans Lehmkuhl,
Ingeborg A Hauser,
Martin Dreyling,
Michael Kneba,
Barbara Gärtner,
Ioannis Anagnostopoulos,
Hanno Riess,
Wolfram Klapper,
Ralf U Trappe
[show abstract]
[hide abstract]
ABSTRACT: Plasmablastic posttransplant lymphoma is a rare subtype of monomorphic B-cell posttransplant lymphoproliferative disorder (PTLD). There is little published clinical data to guide treatment.
The German prospective PTLD registry D2006-2012 records baseline features, treatment, and outcome of rare PTLD subtypes in adults after solid organ transplantation. Treatment is at the discretion of the local physician. Clinical data on the patients in the registry is collected before, during, and at least 4 weeks, 6 months, 12 and 24 months after treatment.
Eight cases of plasmablastic posttransplant lymphoma were reported to the registry until 2011. The majority occurred as late PTLD in male heart transplant recipients. Extranodal manifestations were common in stage I and in stage IV disease. Histological Epstein-Barr virus (EBV) association was confirmed in five of eight cases. MYC/IGH rearrangement was present in two of six patients examined. Although five of eight patients died from early disease progression, we observed that long-term survival can be achieved in localized (2/3) and in disseminated disease (1/5) by immunosuppression reduction (IR) followed by immediate systemic chemotherapy. However, all patients with cytogenetic aberrations and patients with non-EBV-associated PTLD were refractory to IR and to chemotherapy. Chemotherapy parallel to IR was associated with a high rate of infectious complications.
In this series, IR and local therapy were not sufficient to treat plasmablastic posttransplant lymphoma even in localized disease whereas IR and systemic chemotherapy (CHOP-21) could achieve lasting complete remissions. Cytogenetic aberrations and lack of EBV association were linked with poor outcome.
Transplantation 03/2012; 93(5):543-50. · 4.00 Impact Factor
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Martin Dreyling,
Hanneke C. Kluin-Nelemans,
Sílvia Beà,
Elena Hartmann,
Itziar Salaverria,
Grit Hutter,
Patricia Perez-Galan,
Gael Roue, Christiane Pott,
Steven Le Gouill, [......],
Georg Hess,
Francesco Zaja,
Umberto Vitolo,
Michal Szymczyk,
Jan Walewski,
Vincent Ribrag,
Michael Unterhalt,
Olivier Hermine,
Eva Hoster,
for the European Mantle Cell Lymphoma Network
[show abstract]
[hide abstract]
ABSTRACT: Abstract Mantle cell lymphoma (MCL) is a distinct subtype of malignant lymphoma characterized by the chromosomal translocation t(11;14)(q13;q32), resulting in constitutional overexpression of cyclin D1 and cell cycle dysregulation in virtually all cases. Clinically, MCL shows an aggressive clinical course with a continuous relapse pattern and a median survival of only 3–5 years. However, recently a subset of up to 15% long-term survivors has been identified with a rather indolent clinical course. Advanced stage disease is usually apparent already at first clinical manifestation; in general, conventional chemotherapy is only palliative and median duration of remissions is only 1–2 years. In 2000, the European MCL Network (http://www.european-mcl.net) was founded, which consists of 15 national lymphoma study groups supplemented by experts in hematopathology, cytogenetics and molecular genetics. During the last decade, the European consortium has successfully initiated the largest phase III trials in MCL worldwide. In the current study generation, the addition of high-dose Ara-C to a rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP)-like regimen followed by myeloablative consolidation achieved a significant improvement of progression-free survival. Similarly, in elderly patients, rituximab maintenance until progression led to a marked prolongation of remission duration. Emerging strategies include proteasome inhibitors, immune modulatory drugs (IMiDs), mammalian target of rapamycin (mTOR) inhibitors and others, all based on the dysregulated control of cell cycle machinery and impairment of several apoptotic pathways. Combination strategies are currently being investigated in numerous trials, but their introduction into clinical practice and current treatment algorithms remains a challenge. Future strategies will apply individualized approaches according to the molecular risk profile of the patient. At the annual conference in Warsaw, recent results of molecular pathogenesis, analyses of current clinical trials and new study concepts were discussed.
11/2011; 52(12):2226-2236.
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Martin Dreyling,
Hanneke C Kluin-Nelemans,
Sílvia Beà,
Elena Hartmann,
Itziar Salaverria,
Grit Hutter,
Patricia Perez-Galan,
Gael Roue, Christiane Pott,
Steven Le Gouill, [......],
Simon Rule,
Georg Hess,
Francesco Zaja,
Umberto Vitolo,
Michal Szymczyk,
Jan Walewski,
Vincent Ribrag,
Michael Unterhalt,
Olivier Hermine,
Eva Hoster
[show abstract]
[hide abstract]
ABSTRACT: Mantle cell lymphoma (MCL) is a distinct subtype of malignant lymphoma characterized by the chromosomal translocation t(11;14)(q13;q32), resulting in constitutional overexpression of cyclin D1 and cell cycle dysregulation in virtually all cases. Clinically, MCL shows an aggressive clinical course with a continuous relapse pattern and a median survival of only 3-5 years. However, recently a subset of up to 15% long-term survivors has been identified with a rather indolent clinical course. Advanced stage disease is usually apparent already at first clinical manifestation; in general, conventional chemotherapy is only palliative and median duration of remissions is only 1-2 years. In 2000, the European MCL Network ( http://www.european-mcl.net ) was founded, which consists of 15 national lymphoma study groups supplemented by experts in hematopathology, cytogenetics and molecular genetics. During the last decade, the European consortium has successfully initiated the largest phase III trials in MCL worldwide. In the current study generation, the addition of high-dose Ara-C to a rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP)-like regimen followed by myeloablative consolidation achieved a significant improvement of progression-free survival. Similarly, in elderly patients, rituximab maintenance until progression led to a marked prolongation of remission duration. Emerging strategies include proteasome inhibitors, immune modulatory drugs (IMiDs), mammalian target of rapamycin (mTOR) inhibitors and others, all based on the dysregulated control of cell cycle machinery and impairment of several apoptotic pathways. Combination strategies are currently being investigated in numerous trials, but their introduction into clinical practice and current treatment algorithms remains a challenge. Future strategies will apply individualized approaches according to the molecular risk profile of the patient. At the annual conference in Warsaw, recent results of molecular pathogenesis, analyses of current clinical trials and new study concepts were discussed.
Leukemia & lymphoma 08/2011; 52(12):2226-36. · 2.40 Impact Factor
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Anastasia Hadzidimitriou,
Andreas Agathangelidis,
Nikos Darzentas,
Fiona Murray,
Marie-Helene Delfau-Larue,
Lone Bredo Pedersen,
Alba Navarro Lopez,
Antonis Dagklis,
Paul Rombout,
Kheira Beldjord, [......],
Patricia Groenen,
Paolo Ghia,
Birgitta Sander,
Theodora Papadaki,
Elias Campo,
Christian Geisler,
Richard Rosenquist,
Frederic Davi, Christiane Pott,
Kostas Stamatopoulos
[show abstract]
[hide abstract]
ABSTRACT: We examined 807 productive IGHV-IGHD-IGHJ gene rearrangements from mantle cell lymphoma (MCL) cases, by far the largest series to date. The IGHV gene repertoire was remarkably biased, with IGHV3-21, IGHV4-34, IGHV1-8, and IGHV3-23 accounting for 46.3% of the cohort. Eighty-four of 807 (10.4%) cases, mainly using the IGHV3-21 and IGHV4-34 genes, were found to bear stereotyped heavy complementarity-determining region 3 (VH CDR3) sequences and were placed in 38 clusters. Notably, the MCL stereotypes were distinct from those reported for chronic lymphocytic leukemia. Based on somatic hypermutation (SHM) status, 238/807 sequences (29.5%) carried IGHV genes with 100% germ line identity; the remainder (569/807; 70.5%) exhibited different SHM impact, ranging from minimal (in most cases) to pronounced. Shared replacement mutations across the IGHV gene were identified for certain subgroups, especially those using IGHV3-21, IGHV1-8, and IGHV3-23. Comparison with other entities, in particular CLL, revealed that several of these mutations were "MCL-biased." In conclusion, MCL is characterized by a highly restricted immunoglobulin gene repertoire with stereotyped VH CDR3s and very precise SHM targeting, strongly implying a role for antigen-driven selection of the clonogenic progenitors. Hence, an antigen-driven origin of MCL could be envisaged, at least for subsets of cases.
Blood 07/2011; 118(11):3088-95. · 9.90 Impact Factor
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Christiane Pott
[show abstract]
[hide abstract]
ABSTRACT: The prognostic impact of minimal residual disease (MRD) has been demonstrated for several hematologic malignancies. While in acute lymphoblastic leukemias MRD assessment by polymerase chain reaction (PCR)-based methods has been established as an important tool for clinical risk assessment and is part of clinical management, data demonstrating a prognostic value of MRD in mantle cell lymphoma (MCL) were sparse and results from randomized trials have been published only recently. In the present review technical aspects of different MRD detection methods are discussed, as well as the prognostic relevance of MRD in the context of clinical trials in patients with MCL. Furthermore, recommendations are given for workflow and useful implication of MRD in future clinical trials design.
Seminars in Hematology 07/2011; 48(3):172-84. · 3.99 Impact Factor
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[show abstract]
[hide abstract]
ABSTRACT: Autologous stem cell transplantation (autoSCT) has improved the outcome of patients with mantle cell lymphoma (MCL) considerably. However, little is known about the patterns and outcome of MCL recurrence after autoSCT.
The authors conducted a retrospective study of 118 patients with MCL who underwent autoSCT from August 1992 to August 2008 at 3 different referral centers in Germany.
Fifty-two relapses occurred for a cumulative incidence of 46% after 5 years. Only 3 patients relapsed after 5 years (at 90 months, 91 months, and 171 months) after undergoing autoSCT. A Cox regression analysis of the incidence of relapse identified not receiving rituximab before autoSCT and undergoing salvage autoSCT as predictive factors for relapse, whereas cytosine arabinoside intensification; a total body irradiation-based, high-dose regimen; patient age; and year of transplantation had no influence. The median overall survival (OS) after relapse was 23 months. Twenty patients (39%) underwent allogeneic stem cell transplantation (alloSCT) for relapse, and 11 of those patients remained in ongoing complete remission at the time of the current report. It is noteworthy that there were 4 long-term survivors who lived for >5 years after relapse even without undergoing alloSCT. A Cox regression analysis of OS after relapse revealed that the response duration after autoSCT was an adverse predictor of OS, whereas alloSCT was associated with a significantly longer OS after relapse.
The current results indicated that autoSCT was capable of inducing long-term remission up to 16 years after treatment, but the outcome of patients with MCL who relapsed after autoSCT was poor, especially if their response duration after autoSCT was short. However, for a subset of patients with relapsed MCL, alloSCT may offer the possibility of durable survival, and individual patients can enjoy long-term survival after relapse even without undergoing alloSCT.
Cancer 05/2011; 117(9):1901-10. · 4.77 Impact Factor
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Itziar Salaverria,
Claudia Philipp,
Ilske Oschlies,
Christian W Kohler,
Markus Kreuz,
Monika Szczepanowski,
Birgit Burkhardt,
Heiko Trautmann,
Stefan Gesk,
Miroslaw Andrusiewicz, [......],
Julia Richter,
Maciej Rosolowski,
Carsten Schwaenen,
Harald Stein,
Lorenz Trümper,
Swen Wessendorf,
Rainer Spang,
Ralf Küppers,
Wolfram Klapper,
Reiner Siebert
[show abstract]
[hide abstract]
ABSTRACT: The prognosis of germinal center-derived B-cell (GCB) lymphomas, including follicular lymphoma and diffuse large-B-cell lymphoma (DLBCL), strongly depends on age. Children have a more favorable outcome than adults. It is not known whether this is because of differences in host characteristics, treatment protocols, or tumor biology, including the presence of chromosomal alterations. By screening for novel IGH translocation partners in pediatric and adult lymphomas, we identified chromosomal translocations juxtaposing the IRF4 oncogene next to one of the immunoglobulin (IG) loci as a novel recurrent aberration in mature B-cell lymphoma. FISH revealed 20 of 427 lymphomas to carry an IG/IRF4-fusion. Those were predominantly GCB-type DLBCL or follicular lymphoma grade 3, shared strong expression of IRF4/MUM1 and BCL6, and lacked PRDM1/BLIMP1 expression and t(14;18)/BCL2 breaks. BCL6 aberrations were common. The gene expression profile of IG/IRF4-positive lymphomas differed from other subtypes of DLBCL. A classifier for IG/IRF4 positivity containing 27 genes allowed accurate prediction. IG/IRF4 positivity was associated with young age and a favorable outcome. Our results suggest IRF4 translocations to be primary alterations in a molecularly defined subset of GCB-derived lymphomas. The probability for this subtype of lymphoma significantly decreases with age, suggesting that diversity in tumor biology might contribute to the age-dependent differences in prognosis of lymphoma.
Blood 04/2011; 118(1):139-47. · 9.90 Impact Factor
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Leandro Cerchietti,
Christine Damm-Welk,
Inga Vater,
Wolfram Klapper,
Lana Harder, Christiane Pott,
Shao Ning Yang,
Alfred Reiter,
Reiner Siebert,
Ari Melnick,
Willi Woessmann
[show abstract]
[hide abstract]
ABSTRACT: ALK positive diffuse large B-cell lymphomas (DLBCL) are a distinct lymphoma subtype associated with a poor outcome. Most of them feature a t(2;17) encoding a clathrin (CLTC)-ALK fusion protein. The contribution of deregulated ALK-activity in the pathogenesis and maintenance of these DLBCLs is not yet known. We established and characterized the first CLTC-ALK positive DLBCL cell line (LM1). LM1 formed tumors in NOD-SCID mice. The selective ALK inhibitor NVP-TAE684 inhibited growth of LM1 cells in vitro at nanomolar concentrations. NVP-TAE684 repressed ALK-activated signalling pathways and induced apoptosis of LM1 DLBCL cells. Inhibition of ALK-activity resulted in sustained tumor regression in the xenotransplant tumor model. These data indicate a role of CLTC-ALK in the maintenance of the malignant phenotype thereby providing a rationale therapeutic target for these otherwise refractory tumors.
PLoS ONE 01/2011; 6(4):e18436. · 4.09 Impact Factor
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[show abstract]
[hide abstract]
ABSTRACT: The impact of single-nucleotide polymorphisms (SNPs) on tumour susceptibility and pathogenesis has gained enormous attention. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based genotyping facilitates the analysis of short DNA amplicons and is, therefore, a promising tool for the investigation of formalin-fixed paraffin-embedded (FFPE) tissue samples, particularly in targeted genotyping analysis.
To examine the applicability of genotyping FFPE DNA with MALDI-TOF MS in multiplex reactions, we investigated five DNA samples extracted from FFPE tumour specimens from follicular lymphoma patients using different extraction methods (phenol-chloroform, commercial kit). Thirty-one SNPs from 25 genes, integrated in different-sized multiplex assays (7-plex, 10-plex, 14-plex, 24-plex), were analyzed. To investigate the reliability of genotyping tumour-derived DNA extracted from FFPE tissue, we examined 64 FFPE tumour specimens in comparison with matched germline DNA samples.
Call rates of 99.6 (274/275) and 93.5% (257/275) were observed for the DNA extracted with the phenol-chloroform approach or the commercial extraction kit, respectively. Increasing the number of SNPs per assay resulted in reduced genotyping call rates and genotyping quality, especially in the DNA samples isolated with the commercial extraction kit. When comparing the genotypes of DNA derived from germline and tumour (FFPE) specimens, a perfect concordance rate of 100% was detected.
Our data delineate that MALDI-TOF-based genotyping of FFPE DNA is reliable and reproducible even in multiplex reactions, enabling the retrospective investigation of FFPE study cohorts in future experiments.
Pharmacogenetics and Genomics 10/2010; 20(10):598-604. · 3.48 Impact Factor
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Martin Dreyling,
Eva Hoster,
Silvia Bea,
Elena Hartmann,
Heike Horn,
Grit Hutter,
Itziar Salaverria, Christiane Pott,
Marek Trneny,
Steven Le Gouill,
Sergio Cortelazzo,
Michal Szymczyk,
Wojciech Jurczak,
Ofer Shpilberg,
Vincent Ribrag,
Olivier Hermine
[show abstract]
[hide abstract]
ABSTRACT: Mantle cell lymphoma (MCL) is a distinct subtype of malignant lymphoma which is characterized by the chromosomal translocation t(11;14)(q13;q32), resulting in constitutional overexpression of cyclin D1 and cell cycle dysregulation in virtually all cases. Clinically, MCL shows an aggressive clinical course with a continuous relapse pattern and a median survival of only 3-5 years. However, recently a subset of 15% long-term survivors has been identified with a rather indolent clinical course, even after conventional treatment strategies only. Advanced stage disease is usually apparent already at first clinical manifestation; thus, conventional chemotherapy is only palliative, and the median duration of remissions is only 1-2 years. Emerging strategies including proteasome inhibitors, immune modulatory drugs (IMiDs), mTOR inhibitors, and others are based on the dysregulated control of cell cycle machinery and impaired apoptotic pathways. Monotherapy of these compounds achieves efficacy comparable to conventional chemotherapy in relapsed MCL, and combination strategies are currently being investigated in numerous trials; however, their introduction into clinical practice and current treatment algorithms remain a challenge. In 2000 the European MCL Network ( http://www.european-mcl.net ) was founded, consisting of 15 national lymphoma study groups supplemented by experts in histopathology and molecular genetics. During the past decade, the European consortium has successfully initiated the largest phase III trials in MCL worldwide, with a current annual recruitment of almost 200 patients per year in first-line studies. In detail, in prospective randomized studies, the addition of a B-lymphocyte specific antibody doubled the median progression-free survival from 14 to 28 months, and a dose-intensified consolidation with high-dose radiochemotherapy and subsequent autologous stem cell transplant resulted in superior response duration (3.7 vs. 1.6 years) and even improved overall survival in a recent analysis. Future strategies will apply individualized approaches according to the molecular risk profile of the patient. At the recent annual conference in Jerusalem, recent results of molecular pathogenesis, analyses of current clinical trials, and new study concepts were discussed.
Leukemia & lymphoma 09/2010; 51(9):1612-22. · 2.40 Impact Factor
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Christiane Pott,
Eva Hoster,
Marie-Helene Delfau-Larue,
Kheira Beldjord,
Sebastian Böttcher,
Vahid Asnafi,
Anne Plonquet,
Reiner Siebert,
Evelyne Callet-Bauchu,
Niels Andersen, [......],
Marek Trneny,
Jan Walewski,
Peter Dreger,
Michael Unterhalt,
Wolfgang Hiddemann,
Michael Kneba,
Hanneke C Kluin-Nelemans,
Olivier Hermine,
Elizabeth Macintyre,
Martin Dreyling
[show abstract]
[hide abstract]
ABSTRACT: The prognostic impact of minimal residual disease (MRD) was analyzed in 259 patients with mantle cell lymphoma (MCL) treated within 2 randomized trials of the European MCL Network (MCL Younger and MCL Elderly trial). After rituximab-based induction treatment, 106 of 190 evaluable patients (56%) achieved a molecular remission (MR) based on blood and/or bone marrow (BM) analysis. MR resulted in a significantly improved response duration (RD; 87% vs 61% patients in remission at 2 years, P = .004) and emerged to be an independent prognostic factor for RD (hazard ratio = 0.4, 95% confidence interval, 0.1-0.9, P = .028). MR was highly predictive for prolonged RD independent of clinical response (complete response [CR], complete response unconfirmed [CRu], partial response [PR]; RD at 2 years: 94% in BM MRD-negative CR/CRu and 100% in BM MRD-negative PR, compared with 71% in BM MRD-positive CR/CRu and 51% in BM MRD-positive PR, P = .002). Sustained MR during the postinduction period was predictive for outcome in MCL Younger after autologous stem cell transplantation (ASCT; RD at 2 years 100% vs 65%, P = .001) and during maintenance in MCL Elderly (RD at 2 years: 76% vs 36%, P = .015). ASCT increased the proportion of patients in MR from 55% before high-dose therapy to 72% thereafter. Sequential MRD monitoring is a powerful predictor for treatment outcome in MCL. These trials are registered at www.clinicaltrials.gov as #NCT00209222 and #NCT00209209.
Blood 04/2010; 115(16):3215-23. · 9.90 Impact Factor
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Jose I Martin-Subero,
Ole Ammerpohl,
Marina Bibikova,
Eliza Wickham-Garcia,
Xabier Agirre,
Sara Alvarez,
Monika Brüggemann,
Stefanie Bug,
Maria J Calasanz,
Martina Deckert, [......],
José Román-Gómez,
Marc Seifert,
Harald Stein,
Javier Suela,
Lorenz Trümper,
Inga Vater,
Felipe Prosper,
Claudia Haferlach,
Juan Cruz Cigudosa,
Reiner Siebert
[show abstract]
[hide abstract]
ABSTRACT: Alterations in the DNA methylation pattern are a hallmark of leukemias and lymphomas. However, most epigenetic studies in hematologic neoplasms (HNs) have focused either on the analysis of few candidate genes or many genes and few HN entities, and comprehensive studies are required.
Here, we report for the first time a microarray-based DNA methylation study of 767 genes in 367 HNs diagnosed with 16 of the most representative B-cell (n = 203), T-cell (n = 30), and myeloid (n = 134) neoplasias, as well as 37 samples from different cell types of the hematopoietic system. Using appropriate controls of B-, T-, or myeloid cellular origin, we identified a total of 220 genes hypermethylated in at least one HN entity. In general, promoter hypermethylation was more frequent in lymphoid malignancies than in myeloid malignancies, being germinal center mature B-cell lymphomas as well as B and T precursor lymphoid neoplasias those entities with highest frequency of gene-associated DNA hypermethylation. We also observed a significant correlation between the number of hypermethylated and hypomethylated genes in several mature B-cell neoplasias, but not in precursor B- and T-cell leukemias. Most of the genes becoming hypermethylated contained promoters with high CpG content, and a significant fraction of them are targets of the polycomb repressor complex. Interestingly, T-cell prolymphocytic leukemias show low levels of DNA hypermethylation and a comparatively large number of hypomethylated genes, many of them showing an increased gene expression.
We have characterized the DNA methylation profile of a wide range of different HNs entities. As well as identifying genes showing aberrant DNA methylation in certain HN subtypes, we also detected six genes--DBC1, DIO3, FZD9, HS3ST2, MOS, and MYOD1--that were significantly hypermethylated in B-cell, T-cell, and myeloid malignancies. These might therefore play an important role in the development of different HNs.
PLoS ONE 01/2009; 4(9):e6986. · 4.09 Impact Factor
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José I Martín-Subero,
Markus Kreuz,
Marina Bibikova,
Stefan Bentink,
Ole Ammerpohl,
Eliza Wickham-Garcia,
Maciej Rosolowski,
Julia Richter,
Lidia Lopez-Serra,
Esteban Ballestar, [......],
Michael Weber,
Swen Wessendorf,
Markus Loeffler,
Lorenz Trümper,
Harald Stein,
Rainer Spang,
Manel Esteller,
David Barker,
Dirk Hasenclever,
Reiner Siebert
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ABSTRACT: Lymphomas are assumed to originate at different stages of lymphocyte development through chromosomal aberrations. Thus, different lymphomas resemble lymphocytes at distinct differentiation stages and show characteristic morphologic, genetic, and transcriptional features. Here, we have performed a microarray-based DNA methylation profiling of 83 mature aggressive B-cell non-Hodgkin lymphomas (maB-NHLs) characterized for their morphologic, genetic, and transcriptional features, including molecular Burkitt lymphomas and diffuse large B-cell lymphomas. Hierarchic clustering indicated that methylation patterns in maB-NHLs were not strictly associated with morphologic, genetic, or transcriptional features. By supervised analyses, we identified 56 genes de novo methylated in all lymphoma subtypes studied and 22 methylated in a lymphoma subtype-specific manner. Remarkably, the group of genes de novo methylated in all lymphoma subtypes was significantly enriched for polycomb targets in embryonic stem cells. De novo methylated genes in all maB-NHLs studied were expressed at low levels in lymphomas and normal hematopoietic tissues but not in nonhematopoietic tissues. These findings, especially the enrichment for polycomb targets in stem cells, indicate that maB-NHLs with different morphologic, genetic, and transcriptional background share a similar stem cell-like epigenetic pattern. This suggests that maB-NHLs originate from cells with stem cell features or that stemness was acquired during lymphomagenesis by epigenetic remodeling.
Blood 01/2009; 113(11):2488-97. · 9.90 Impact Factor
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Inga Vater,
Florian Wagner,
Markus Kreuz,
Hilmar Berger,
José I Martín-Subero, Christiane Pott,
Jose A Martinez-Climent,
Wolfram Klapper,
Kristina Krause,
Martin J S Dyer,
Stefan Gesk,
Lana Harder,
Alberto Zamo,
Martin Dreyling,
Dirk Hasenclever,
Norbert Arnold,
Reiner Siebert
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ABSTRACT: The translocation t(11;14)(q13;q32) is the genetic hallmark of mantle cell lymphoma (MCL) but is not sufficient for inducing lymphomagenesis. Here we performed genome-wide 100K GeneChip Mapping in 26 t(11;14)-positive MCL and six MCL cell lines. Partial uniparental disomy (pUPD) was shown to be a recurrent chromosomal event not only in MCL cell lines but also in primary MCL. Remarkably, pUPD affected recurrent targets of deletion like 11q, 13q and 17p. Moreover, we identified 12 novel regions of recurrent gain and loss as well as 12 high-level amplifications and eight homozygously deleted regions hitherto undescribed in MCL. Interestingly, GeneChip analyses identified different genes, encoding proteins involved in microtubule dynamics, such as MAP2, MAP6 and TP53, as targets for chromosomal aberration in MCL. Further investigation, including mutation analyses, fluorescence in situ hybridisation as well as epigenetic and expression studies, revealed additional aberrations frequently affecting these genes. In total, 19 of 20 MCL cases, which were subjected to genetic and epigenetic analyses, and five of six MCL cell lines harboured at least one aberration in MAP2, MAP6 or TP53. These findings provide evidence that alterations of microtubule dynamics might be one of the critical events in MCL lymphomagenesis contributing to chromosomal instability.
British Journal of Haematology 12/2008; 144(3):317-31. · 4.94 Impact Factor
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Inga Nieländer,
José I Martín-Subero,
Florian Wagner,
Michael Baudis,
Stefan Gesk,
Lana Harder,
Dirk Hasenclever,
Wolfram Klapper,
Markus Kreuz, Christiane Pott,
José A Martinez-Climent,
Martin Dreyling,
Norbert Arnold,
Reiner Siebert
Haematologica 07/2008; 93(6):949-50. · 6.42 Impact Factor
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Sebastian Böttcher,
Matthias Ritgen,
Sebastian Buske,
Stefan Gesk,
Wolfram Klapper,
Eva Hoster,
Wolfgang Hiddemann,
Michael Unterhalt,
Martin Dreyling,
Reiner Siebert,
Michael Kneba, Christiane Pott
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ABSTRACT: The increasing application of multi-color flow cytometry assays for staging and follow-up in mantle cell lymphoma necessitates that the specificity and sensitivity of this technique are evaluated. Data from prospective clinical trials comparing the clinical applicability of flow cytometry to routine diagnostic methods and to polymerase chain reaction are currently lacking.
We applied a standardized four-color flow cytometry assay to 281 prospectively collected peripheral blood and bone marrow samples from 98 patients with mantle cell lymphoma participating in a multi-center clinical trial and compared the results to those obtained with conventional clinical staging and consensus primer IGH-polymerase chain reaction.
The maximum sensitivity of flow cytometry using light chain restriction in CD19+CD5+ subpopulations was 8.0 x 10(-4) while flow cytometry that relied on immunophenotypic aberrations was less sensitive (2.4 x 10(-3)). Mantle cell lymphoma cells were detected in 87.3% of 110 pre-treatment samples from 84 patients by flow cytometry and in 94.5% by polymerase chain reaction. Eight out of 84 patients (9.5%) diagnosed clinically as having stage II or III disease showed peripheral blood or bone marrow involvement according to flow cytometry, thus documenting more advanced disease. At follow-up residual lymphoma cells were detected by flow cytometry and concordantly by polymerase chain reaction in 10/171 samples (5.8%); however, 31 follow-up samples (18.1%) were positive for minimal residual disease according only to polymerase chain reaction analysis.
The sensitivity of four-color flow cytometry is comparable to that of IGH-polymerase chain reaction at initial staging but is less sensitive at follow-up after immuno-chemotherapy. Both techniques are highly valuable methods for accurate initial staging.
Haematologica 05/2008; 93(4):551-9. · 6.42 Impact Factor
