Molecular Genetics of Myeloid Leukemia

Contact/Address

Prof. Dr. med. Konstanze Döhner
Department of Internal Medicine III
University Hospital of Ulm
Albert-Einstein-Allee 23
89081 Ulm
Germany
Phone: +49-(0)731-500-45543
Fax: +49-(0)731-500-45505

Technicians

Nuray Atasoy

Oliva Gallegos Cossio

Martina Enz

Jennifer Guhl

Judith Kühnemann

Susanne Kuhn

Michael Ruhland

Heidi Schmidt

Silvia Schuhhardt

Olesia Sirik

Sabrina Skambraks

(A) Identification of novel gene mutations in AML using next-generation based techniques and evaluation of their prognostic impact

Acute myeloid leukemia (AML) is a clonal hematopoietic stem cell malignancy characterized by an accumulation of immature progenitor cells arrested in their differentiation abilities leading to suppression of hematopoiesis. The disease is highly heterogeneous with respect to morphology, immunophenotype, (cyto)genetic and epigenetic signatures as well as treatment response and treatment outcome.

In recent years, there has been an unprecedented rapid advance in our understanding of the molecular and phenotypic diversity of AML. This knowledge has culminated in the identification of pathologic intracellular pathways critical for AML growth and various molecular and immunologic therapeutic targets.

The aim of our studies is to perform comprehensive gene mutation analyses in the context of our ongoing clinical trials. All molecular techniques, including next-generation sequencing techniques, are established in our laboratory. For our projects we take advantage of one of the largest biobank of primary leukemia samples worldwide. With our work we have been able to provide seminal contributions to our current understanding of malignant myeloid disease.

(B) Molecular Monitoring of Measurable Residual Disease (MRD) in AML

MRD assessment in AML is used to provide a quantitative methodology to establish a deeper remission status; to refine post-remission relapse risk assessment, to identify impending relapse to enable early intervention, and as a surrogate endpoint to accelerate drug testing and approval.

Currently, the two most extensively evaluated methodologies are multiparameter flow cytometry-based MRD (MFC-MRD) and molecular MRD (Mol-MRD) assessed by quantitative PCR (qPCR). Emerging exploratory technologies are next-generation sequencing (NGS) and digital PCR.

Leukemia-related abnormalities suitable for qPCR monitoring include mutated NPM1; CBFB::MYH11, RUNX1::RUNX1T1, KMT2A::MLLT3, DEK::NUP214, and BCR::ABL1 gene fusions. If using NGS, error-corrected targeted panel-based approaches are being used. Currently, NGS-based strategies lack standardization as a stand-alone technique for MRD assessment.

All techniques are established in our group and monitoring of MRD is implemented in all our ongoing clinical trials for AML.

• Cell culture
• Fluoresence in situ hybridization (FISH)
• Fluorescence microscopy
• DNA/RNA preparation
• PCR techniques including quantitative real-Time RT-PCR (Taq Man, Light Cycler)
• DNA cloning
• Southern-, Northern-, Western blotting
• DNA sequencing
• Sequence analysis
• Genescan analysis
• DHPLC-based mutation screening (WAVE System)
• LOH analysis (SNP microarrays)
• Gene expression profiling using different platforms (cDNA, Affymetrix)
• Profiling miRNA expression (oligo microarray, real-time PCR)

• Medizinische Fakultät der Universität Ulm („Bausteinprogramm“)
• Deutsche Forschungsgemeinschaft (DFG)
• Bundesministerium für Bildung und Forschung (BMBF)
• European Commision
• Else Kröner-​Fresenius-Stiftung
• Biotechnologische und Pharmazeutische Industrie

Representative publications can be found in the entire list of publications.

Prof. Lars Bullinger
Charité Universitätsmedizin, Berlin

Prof. Hans-​Jörg Fehling
Department of Molecular Immunology, University of Ulm

Prof. Christoph Plass
DKFZ, Department of Molecular Genetics, Heidelberg

Prof. Karsten Rippe
DKFZ, Department of Molecular Genetics, Heidelberg