Mutations in MPN: Hereditary Predispositions, Disease Initiators and Drivers of Progression

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1 Mutations in MPN: Hereditary Predispositions, Disease Initiators and Drivers of Progression Robert Kralovics, PhD CeMM - Center for Molecular Medicine of the Austrian Academy of Sciences Vienna, Austria

2 Myeloproliferative Neoplasms

3 MPN: All Roads Lead to JAK-STAT

4 Disease causing mutations in MPN

5 Triple negative MPN

6 Genetic changes in MPN therapy phenotype diagnosis Acute leukemia Myelodysplastic Normal Myeloproliferative Germline factors JAK2-GGCC TERT MECOM, MYB RBBP6 LNK ATG2B GSKIP del-il1rap Clonal drivers del20q del13q TET2 DNMT3A Disease causing JAK2 MPL CALR Progression associated TP53/MDM4 PRC2 complex ASXL1 CUX1 RUNX1 Adaptation to therapy

7 Genome-wide association study for MPN susceptibility loci

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9 Familial MPN caused by gain-of-function mutations of RBBP6 Linkage analysis Exome sequencing Harutyunyan et al., in preparation

10 Cumulative effect of TERT and JAK2-GGCC

11 Types of predispositions to MPN MPN families GGCC haplotype

12 Genetic changes in MPN therapy phenotype diagnosis Acute leukemia Myelodysplastic Normal Myeloproliferative Germline factors JAK2-GGCC TERT RBBP6 LNK ATG2B GSKIP del-il1rap Clonal drivers del20q del13q TET2 DNMT3A Disease causing JAK2 MPL CALR Progression associated TP53/MDM4 PRC2 complex ASXL1 CUX1 RUNX1 Adaptation to therapy

13 Models of leukemic transformation in MPN Mono-clonal Bi-clonal JAK2-V617F JAK2-V617F TP53 RUNX1 FLT3/NPM1/NRAS JAK2-V617F positive AML JAK2-V617F negative AML de novo like

14 Cytogenetic lesions in MPN disease progression Chronic MPN Acc. phase Leukemic phase Klampfl et al., Blood, 2011

15 Exome sequencing in post-mpn AML T cells as the control tissue Mutations causing MPN Chronic phase MPN Chronic phase sample 14 x Accelerated phase MPN Accelerated phase sample Mutations causing disease progression Mutations causing leukemic transformation Exome sequencing Leukemic sample Secondary AML Elisa Rumi, Mario Cazzola Tiina Berg, Klaudia Bagienski RNA-seq for fusions detection

16 Clonal reconstruction in post-mpn AML

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19 Overlap of mutations and chromosomal lesions FOXP1 V515A CUX1 A802D CUX1

20 Whole transcriptome analysis

21 Fusion event landscape of MPN

22 RNA-seq: Phenotype signatures

23 Variant calling: TruSight and RNA-seq

24 ET PMF PV

25 Frequently mutated genes in spliceosome

26 What can we use mutation profiles for? Diagnosis Estimation of mutational complexity (number/type) Risk stratification, prognosis Estimation of disease stage, early warning of transformation (???) Prediction of treatment response Evaluation of molecular response during treatment (burden quantification) Post treatment: detection of MRD, detection of relapse

27 Summary/Take home messages More than 90% of MPN driven by JAK2, CALR and MPL mutations Triple-negative MPN: atypical MPL and JAK2 mutations, 50% polyclonal germline JAK2 and MPL in many cases Recurrence of individual gene defects is low Cytogenetic lesions have to be considered together with point mutations Gene fusions are rare in MPN Patients display complex mutational profiles esp. during progression Each patient at leukemic phase has a unique mutational profile

28 Genetic changes in MPN therapy phenotype diagnosis Acute leukemia Myelodysplastic Normal Myeloproliferative Germline factors JAK2-GGCC TERT RBBP6 LNK ATG2B GSKIP del-il1rap Clonal drivers del20q del13q TET2 DNMT3A Disease causing JAK2 MPL CALR Progression associated TP53/MDM4 ASXL1 EZH2 CUX1 RUNX1 Adaptation to therapy

29 Acknowledgements CeMM RK lab Tiina Berg Ashot Harutyunyan Thorsten Klampfl Ana Puda Jelena Milosevic Doris Chen Klaudia Bagienski Ciara Cleary Nicole Them Fiorella Schischlik Roland Jäger Damla Olcaydu CeMM Denise Barlow Sebastian Nijman Giulio Superti-Furga Jacques Colinge Keiryn Bennett Christoph Bock IMP Johannes Zuber Meinrad Busslinger Med.Univ. Vienna Heinz Gisslinger Ulrich Jäger Peter Valent University of Pavia Mario Cazzola Francesco Passamonti Daniela Pietra Elisa Rumi University of Florence Alessandro Vannucchi Lorenzo Tozzi Masaryk University Brno Miroslav Penka Michael Doubek Sarka Pospisilova Zdenek Racil SFB28 Veronika Sexl, VetMedUni Richard Moriggl, LBI-CR Mathias Mueller, VetMedUni INSERM Sylvie Hermouet, Nantes Jean-Jacques Kiladjian, Paris Christine Bellanne-Chantelot, Paris William Vainchenker, Paris Jean-Luc Villeval, Paris University of Birmingham Jon Frampton Paloma Garcia University of Southampton Nick Cross University of Belgrade Sonja Pavlovic Natasa Tosic University Hospital Innsbruck Michael Steurer

30 Questions 1. Mutations in which genes are relevant for the diagnosis of MPNs? A. JAK2, MPL B. JAK2, CALR, MPL C. JAK2, ASLX1 2. Which of the following gene mutations you may detect at MPN progression? A. JAK2-V617F B. RUNX1, ASLX1, EZH2, TP53 C. TET2, DNMT3A