Michael N. Dworzak Group

Michael N Dworzak, email: michael.dworzak(at)ccri.at

Immunological Diagnostics

We develop new diagnostic methods for children and adolescents with leukemia and lymphomas using flow cytometry immunophenotyping.

Our working group’s research focus deals with the development and validation of new diagnostic methods based on flow cytometry. We focus mainly on leukemia and lymphomas, which make up about 50% of all cases of cancer in children and adolescents. The topics of our work are investigations into disease-associated peculiarities of protein expression, which could in future be exploited clinically for elaborate diagnostics, risk stratification and treatment tailoring to individual needs. Our scientific focus also lies on the goal of developing new therapies for children and adolescents with leukemia and lymphomas.

Team

Our research in more detail

(1) Standardization and quality control of diagnostics (Immunophenotyping and MRD in ALL and AML) in an international context

Standardization of immunophenotyping as well as of FCM-MRD assessment in ALL and AML is necessary to allow concordant multicentric application of the methodologies. We already induced an international extensive standardization and validation effort for immunophenotyping as well as MRD-assessment in ALL and elaborated common guidelines. We are currently working intensively on the standardization of the AML-MRD methodology in a broad international context (see pt. 2). We are constantly applying comprehensive training and QC programs involving a large number of international laboratories within the I-BFM consortium to complement standardization of the methodology with an educational component as well as with persistent quality control measures. These QC measures include twinning-maturation programs, obligatory participation in external QC programs, organizing FCM-data file ring robin rounds, independent data surveys on study results as well as regular joint educational feedback meetings.

(2) Flow cytometric minimal residual disease assessment (FCM-MRD) in AML

FCM-MRD has been demonstrated useful for refined identification of AML patients at high risk of relapse by some research groups including the Italian pedAML consortium. However, the use of non-standardized protocols including different staining panels, analysis and gating strategies etc. hampers a broad implementation of FCM-MRD monitoring in clinical routine. We are therefore currently inducing an extensive and broad standardization program within the EuPAL-FLOW (European Pediatric Acute Leukemia) network, containing the majority of reference laboratories of the three major European consortia (AIEOP-BFM, NOPHO-DBH, MyeChild01). Besides standardization, another aim of the EuPAL network is to prospectively acquire and validate large FCM-MRD data sets in prospective and multi-center clinical studies. We also compare our FCM-MRD methodology with another independent methodology (DNA-PCR) as well as with morphology and correlate the results with the outcome of the patients to identify the method that correlates best with the outcome of the patients.

(3) Automation of MRD assessment in ALL and AML using machine learning

In close collaboration with the TU Vienna (Computer Vision Lab) we have already developed a software for automated detection and quantification of MRD in ALL (“AutoFlow”). In contrast to ALL, MRD detection and quantification in AML is much more complicated due to the much more complex pathology of AML. Hence, a reliable and objective tool for automated FCM analysis would foster standardization of the methodology and allow quality-assured clinical follow-up assessment of AML-MRD. In our current project “MyeFlow” we are using the preliminary work of our project “FlowCluster”, where we have collected a large database of FCM-MRD data as well as developed software prototypes. We continuously extend the database throughout the current project and integrate as well as enhance the developed algorithms to develop a clinical decision support system and to enable its use in the clinical practice of AML.

(4) Functional precision oncology pediatric AML

Constitutive activation of signaling pathways in AML cells is associated with poor prognosis. Hence, signal pathway interception in addition to chemotherapy regimen has emerged as a novel approach for the treatment of AML. Several new drugs have recently entered the clinics for the treatment of AML in adults, but their efficacy is poorly understood in pedAML. We employ an innovative long-term 3D culture approach as well as phospho-profiling technology to study the efficacy of selected novel signal interceptors/drugs in different subtypes of pedAML to develop a tool for personalized treatment of pedAM. Together with the group of K. Boztug we integrate FCM-based characterization of AML (including phospho-profiling) augmented with ex-vivo chemosensitivity testing using image-based methods and deep genomic characterization of AML to define functional subgroups of pedAML and to identify patients at risk for therapy resistance/relapse as well as to define potential drugs for targeted treatment approaches.

Funding

EU - Marie Curie FP7-People-2013-IAPP Nr. 610872, project "AutoFLOW - Automation of Flow Cytometric Analysis for Quality-Assured Follow-up Assessment to Guide Curative Therapy for Acute Lymphoblastic Leukemia in Children"

Vienna Business Agency, ZIT 13, ID 1207843, project "flowCLUSTER - Automated MRD-assessment in AML"

Vienna Business Agency, Science to Products Call 2019, ID 2841342, project "MyeFLOW - Automated MRD-assessment in AML"

Linking Research and Patients´ Needs Call 2007 Nr. LS07-037, project "Flow cytometric signal typing for therapy response"

Most important collaboration partners

Name, institution, country	Topic
Prof. Buldini, Univ. Padova, Italy	Immunophenotyping in pedAL
Prof. Reinhardt, Univ. Essen, Germany	AML precision oncology and MRD studies
Dr. Tierens, NOPHO group	AML-FLOW-MRD
Prof. Locatelli, Univ. Rome, HPBG, Italy	AML precision oncology and MRD studies
Prof. Kiss, Univ. Debrecen, Hungary	ALL-FLOW-MRD within ALL-IC-BFM trials

Selected Articles

A. Schumich°, M. Prchal-Murphy°, M. Maurer-Granofszky, A°. Hoelbl-Kovacic°, N. Muhlegger, U. Potschger, S. Fajmann, O.A. Haas, K. Nebral, N. von Neuhoff, M. Zimmermann, H. Boztug, M. Rasche, M. Dolezal, C. Walter, D. Reinhardt, V. Sexl*, and M.N. Dworzak*, Phospho-Profiling Linking Biology and Clinics in Pediatric Acute Myeloid Leukemia. Hemasphere, 2020. 4(1): p. e312. °equally contributed as first authors, * equally contributed as corresponding authors
B. Buldini, M. Maurer-Granofszky, E. Varotto, M.N. Dworzak. Flow-cytometric monitoring of minimal residual disease in pediatric patients with acute myeloid leukemia: Recent advances and future strategies. Front Pediatr. 2019 Oct 11;7:412.doi: 10.3389/fped.2019.00412. eCollection 2019
M. Reiter°, M. Diem°, A. Schumich°, M. Maurer-Granofszky°, L. Karawajew, J.G. Rossi, R. Ratei, S. Groeneveld-Krentz, E.O. Sajaroff, S. Suhendra, M. Kampel, M.N. Dworzak, International Berlin-Frankfurt-Munster (i-BFM) Flow network and the AutoFlow consortium, Automated Flow Cytometric MRD Assessment in Childhood Acute B- Lymphoblastic Leukemia Using Supervised Machine Learning. Cytometry A, 2019. 95(9): p. 966-975. °equally contributed as first authors
D. Schinnerl°, E. Mejstrikova°, A. Schumich°, M. Zaliova, K. Fortschegger, K. Nebral, A. Attarbaschi, K. Fiser, M.O. Kauer, N. Popitsch, S. Haslinger, A. Inthal, B. Buldini, G. Basso, J.P. Bourquin, G. Gaipa, M. Brüggemann, T. Feuerstein, M. Maurer-Granofszky ,R. Panzer-Grümayer, J. Trka, G. Mann, O.A. Haas, O. Hrusak, M.N. Dworzak*, S. Strehl°. CD371 cell surface expression: a unique feature of DUX4-rearranged acute lymphoblastic leukemia. Haematologica. 2019 Aug;104(8):e*52-e355. doi: 10.3324/haematol.2018.214353. Epub 2019 Jan 31.Haematologica. 2019. PMID: 30705095. °equally contributed as first authors, * equally contributed as corresponding authors
M. N. Dworzak, B. Buldini, G. Gaipa, R. Ratei, O. Hrusak, D. Luria, E. Rosenthal, J.P. Bourquin, M. Sartor, A. Schumich, L. Karawajew, E. Mejstrikova, O. Maglia, G. Mann, W.D: Ludwig, A. Biondi, M. Schrappe, G. Basso; International-BFM-FLOW-network. AIEOP-BFM consensus guidelines 2016 for flow cytometric immunophenotyping of Pediatric acute lymphoblastic leukemia. Cytometry B Clin Cytom. 2018 Jan;94(1):82-93. doi: 10.1002/cyto.b.21518. Epub 2017 Feb 21.Cytometry B Clin Cytom. 2018. PMID: 28187514

About Michael Dworzak

Univ-Prof. Dr. Michael N. Dworzak (MD) is vice chair of the St. Anna Kinderspital in Vienna and Section Head of pediatric Oncology & Hematology. He is also chair of the Austrian pediatric AML-BFM clinical trial group, co-chair of the international AML-BFM study group and head of the Immunological Diagnostics laboratory at CCRI and Labdia Labordiagnostik GmbH. This laboratory unit is the national reference diagnostic center for immunophenotyping and flow cytometric MRD-evaluation for pediatric leukemia in Austria. He coordinates several international FLOW study groups and networks, including the iBFM FLOW network as well as the EuPAL FLOW Diagnostic Network. His major achievements include the establishment, clinical validation, and international dissemination of an innovative landmark-technology for response assessment in pediatric leukemias based on flow-cytometric minimal residual detection (FLOW-MRD). This eventually led to the integration of FLOW-MRD into clinical treatment protocols, which are applied by an intercontinental consortium for stratification of pediatric patients into relapse-risk-based treatment strata worldwide.