Molecular Pathogenesis and Progression of Lymphoproliferative Disorders- Universitätsklinikum Ulm

Molecular Pathogenesis and Progression of Lymphoproliferative Disorders

Contact/Address

Prof. Dr. Stephan Stilgenbauer

Department of Internal Medicine III
University of Ulm
Albert-Einstein-Allee 23
89081 Ulm
Germany

E-Mail: stephan.stilgenbauer[at]uniklinik-ulm.de

Phone (direct):
Phone (study office):
Fax:

0049+(0)731-500-45548
0049+(0)731-500-45901
0049+(0)731-500-45905

Project Leader

Prof. Dr. Stephan Stilgenbauer

Scientific Lab Members

Dr. Johannes Bloehdorn
Dr. Andreas Bühler
Dr. Jennifer Edelmann
Dr. Daniela Opel
Dr. Andrea Schnaiter
Dr. Dirk Winkler

Technicians

Alexandra Brodbeck
Christina Galler
Katharina Höger
Sabrina Rau
Sabrina Kless
Katja Scheibel
Doris Winter

Research Fields

(A)

Incidence of genomic aberrations of CLL and mantle cell lymphoma (MCL). Analysis by fluorescence in-situ hybridization (FISH) with comprehensive probe sets for +(3q26), del(6q21), +(8q24), del(11q22-23), +(12q13), del(13q14), t(11;14)(q13;q32), t(14;18)(q32;q21), t(14q32), del(17p13) in CLL and in addition for del(1p22), del(6q27), +(7p15), del(8p22), del(9p21), del(10p15), +(15q23), del(18p11), +(Xq28) in MCL. Development of automated high-resolution genotyping through microarray based matrix-CGH and SNP-chip analyses.

(B)

Characterization of critical regions and isolation of candidate genes involved in pathogenesis and clinical progression (see also link: Prof. Lichter). For the most frequently involved genomic regions such as del(1p22), +(3q26), del(6q21), del(8p22), +(8q24), del(11q22-23), +(12q13), and del(13q14) the affected genes are mostly unknown. The critical regions are defined and candidate genes are isolated and subjected to expression as well as mutation analyses.

(C)

Identification of VDJ rearrangement and IgVH mutation status in lymphoproliferative disorders. Amplification of VDJ rearrangements from RNA and DNA by GeneScan technology based multiplex PCR and sequencing of the products. Characterization of VDJ structure, VH gene usage and mutation status in various B-cell malignancies to define their role in pathogenesis and clinical course of the disease.

(D)

Gene expression profiling of lymphoproliferative disorders. Deregulation of gene expression (mRNA and miR) is studied through microarray and TaqMan approaches in relation to disease subgroups defined by molecular and clinical markers. Changes in gene expression are studied during disease evolution and under specific treatments. Protein expression of candidate genes, in particular of cell cycle, apoptosis control are studied by Western blot analysis.

(E)

Expression analysis of surrogate markers of pathogenic and prognostic significance in CLL such as ZAP-70, SEP-10, LPL, etc. by flow cytometry, TaqMan, and Western blotting. Technical development and validation of the different markers in well characterized cohorts of patients.

(F)

Mutation profiling of lymphoproliferative disorders. Mutations of different genes (e.g. TP53, ATM, MDM2, etc.) involved in the pathogenesis are studied by DHPLC (Denaturing High Performance Liquid Chromatography) and direct sequencing. The results are analysed in relation to disease subgroups defined by molecular and clinical markers. The consequences of gene mutations are characterised by the analysis of changes or signatures in gene expression, protein expression and functional assays after irradiation or chemotherapy.

(G)

Central genetics reference laboratory of the German CLL Study Group (GCLLSG) focused on characterization of genomic aberrations, mutations, gene expression, cell cycle/apoptosis and tumor bank. Development of databases and compilation of datasets for comprehensive biologic and clinical characterization of leukemia and lymphoma. Identification of molecular risk markers predicting progression, treatment response, and survival. Development and coordination of clinical trial protocols based on molecular risk-assessment with innovative treatment approaches (risk stratification, antibody therapy, stem cell transplant) (CLL2H, CLL3C and CLL.

Techniques

Isolation of tumor cells by ficoll gradient and immunomagnetic cell sorting
fluorescence in-situ hybridization (FISH), also in combination with immunostaining
DNA and RNA preparation of cells and tissues
PCR and RT-PCR techniques, (including multiplex and TaqMan analyses)
DNA sequencing, sequence analysis cloning and preparation of various vector systems (plasmids, cosmids, BAC, PAC, YAC)
microarray expression (mRNA and miR) and genomic profiling (SNP chip).
Flow cytometry (FACS) of diverse intracellular antigens (ZAP-70 etc.)
Sensitive mutation profiling of different genes (e.g. TP53, ATM, MDM2, etc.) by DHPLC (WAVE)

Grants/Funding

Medizinische Fakultät Universität Ulm „Bausteinprogramm“(Research field A, B and C)
Deutsche Krebshilfe (Research field A, B and G)
Wilhelm Sander-Stiftung (Research field B, C and G)
Bundesministerium für Bildung und Forschung (Research field A, B and D)
European Commission (Research field B and D)
Else Kröner - Fresenius Stiftung (Research field C and G)
DFG (research field D)
Deutsche José Carreras Leukämie-Stiftung (research field E and G)

Representative Publications

1:
Zenz T, Mertens D, Küppers R, Döhner H, Stilgenbauer S.
From pathogenesis to treatment of chronic lymphocytic leukaemia.
Nat Rev Cancer. 2010 Jan;10(1):37-50.

2:
Stilgenbauer S, Zenz T, Winkler D, Bühler A, Schlenk RF, Groner S, Busch R, Hensel M, Dührsen U, Finke J, Dreger P, Jäger U, Lengfelder E, Hohloch K, Söling U, Schlag R, Kneba M, Hallek M, Döhner H; German Chronic Lymphocytic Leukemia Study Group.
Subcutaneous alemtuzumab in fludarabine-refractory chronic lymphocytic leukemia: clinical results and prognostic marker analyses from the CLL2H study of the German Chronic Lymphocytic Leukemia Study Group.
J Clin Oncol. 2009 Aug 20;27(24):3994-4001.

3:
Zenz T, Mohr J, Eldering E, Kater AP, Bühler A, Kienle D, Winkler D, Dürig J, van Oers MH, Mertens D, Döhner H, Stilgenbauer S.
miR-34a as part of the resistance network in chronic lymphocytic leukemia.
Blood. 2009 Apr 16;113(16):3801-8.

4:
Zenz T, Kröber A, Scherer K, Häbe S, Bühler A, Benner A, Denzel T, Winkler D, Edelmann J, Schwänen C, Döhner H, Stilgenbauer S.
Monoallelic TP53 inactivation is associated with poor prognosis in chronic lymphocytic leukemia: results from a detailed genetic characterization with long-term follow-up.
Blood. 2008 Oct 15;112(8):3322-9.

5:
Roos G, Kröber A, Grabowski P, Kienle D, Bühler A, Döhner H, Rosenquist R, Stilgenbauer S.
Short telomeres are associated with genetic complexity, high-risk genomic aberrations, and short survival in chronic lymphocytic leukemia.
Blood. 2008 Feb 15;111(4):2246-52.

6:
Kienle D, Katzenberger T, Ott G, Saupe D, Benner A, Kohlhammer H, Barth TF, Höller S, Kalla J, Rosenwald A, Müller-Hermelink HK, Möller P, Lichter P, Döhner H, Stilgenbauer S.
Quantitative gene expression deregulation in mantle-cell lymphoma: correlation with clinical and biologic factors.
J Clin Oncol. 2007 Jul 1;25(19):2770-7.

Collaborations (selection):

Prof. Dr. rer. nat. P. Lichter, DKFZ, Heidelberg
Dipl. Stat. A. Benner, DKFZ, Heidelberg
Dr. R. Rosenquist, Uppsala, Sweden
Dr. N. Chiorazzi, New-York
Dr. R. Dalla-Favera, New-York
Prof. Dr. Ulrich Jäger, Wien
Prof. Dr. P. Möller, Ulm
Prof. Dr. H. K. Müller-Hermelink, Würzburg
Deutsche Krebshilfe "Molekulare Mechanismen bei malignen Lymphomen"
German CLL Study Group (GCLLSG)
European Leukemia Net (ELN)
European Mantle Cell Lymphoma Network
European Research Initiative on CLL (ERIC)