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A Prominent Couple Citation for published version (APA): Ratz, L. (2017). A Prominent Couple: the TMPRSS2:ERG Gene Fusion in Aggressive Prostate Cancer . Datawyse / Universitaire Pers Maastricht. https://doi.org/10.26481/dis.20171215lr Document status and date: Published: 01/01/2017 DOI: 10.26481/dis.20171215lr Document Version: Publisher's PDF, also known as Version of record Please check the document version of this publication: • A submitted manuscript is the version of the article upon submission and before peer-review. There can be important differences between the submitted version and the official published version of record. People interested in the research are advised to contact the author for the final version of the publication, or visit the DOI to the publisher's website. • The final author version and the galley proof are versions of the publication after peer review. • The final published version features the final layout of the paper including the volume, issue and page numbers. Link to publication General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal. If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license above, please follow below link for the End User Agreement: www.umlib.nl/taverne-license Take down policy If you believe that this document breaches copyright please contact us at: [email protected] providing details and we will investigate your claim. Download date: 05 Oct. 2021 A Prominent Couple The TMPRSS2:ERG Gene Fusion in Aggressive Prostate Cancer A Prominent Couple – The TMPRSS2:ERG Gene Fusion in Aggressive Prostate Cancer Leonie Ratz ISBN 978 94 6159 776 2 UMP Cover design and layout by Leonie Ratz UNIVERSITAIRE Printed by Datawyse | Universitaire Pers Maastricht PERS MAASTRICHT Copyright © Leonie Ratz, Cologne 2017 All rights reserved. No parts of this thesis may be reproduced, stored or transmitted in any form or by any means without prior written permission of the author. Cover illustration: collection of silhouettes of men (from shutterstock.inc) A Prominent Couple The TMPRSS2:ERG Gene Fusion in Aggressive Prostate Cancer DISSERTATION to obtain the degree of Doctor at Maastricht University, on the authority of the Rector Magnificus, Prof. Dr. Rianne M. Letschert in accordance with the decision of the Board of Deans, to be defended in public on Friday, 15th of December 2017, at 13:30 hours by Leonie Ratz born on 23rd of March 1987 in Cologne SUPERVISORS Prof. Dr. F. C. S. Ramaekers Prof. Dr. H. Sültmann (German Cancer Research Center and National Center of Tumor Diseases, Heidelberg) CO-SUPERVISORS Privatdozentin Dr. S. M. Klauck (German Cancer Research Center, Heidelberg) Prof. Dr. P. Altevogt (German Cancer Research Center, Heidelberg) ASSESSMENT COMMITTEE Prof. Dr. M. van Engeland (chair) Prof. Dr. J. Schalken (Radboud University Medical Center, Nijmegen) Prof. Dr. S. Pahernik (Nuremberg General Hospital, Paracelsus Medical University, Nuremberg) Prof. Dr. M. Vooijs FUNDING This project was supported through intramural funding by the German Cancer Research Center. CONTENTS List of abbreviations 6 General introduction 9 Chapter 1 The biology of aggressive prostate cancer: Implications for innovative diagnostics and therapy 17 Chapter 2 TMPRSS2:ERG gene fusion variants induce TGF-β signaling and epithelial to mesenchymal transition in human prostate cancer cells 39 Chapter 3 TMPRSS2:ERG overexpression induces changes in the epigenetic signature of human prostate cancer cells: Hypomethylation correlates with upregulation of FZD4 and HLA-DMB 73 Chapter 4 INSM1 induces a neuroendocrine phenotype in prostate cancer cells 107 Chapter 5 General discussion 169 Valorisation 177 Summary 181 Nederlandse samenvatting 187 Deutsche Zusammenfassung 193 Acknowledgements 199 Curriculum vitae 201 Publication list 202 LIST OF ABBREVIATIONS ACPP acid phosphatase, prostate ADT androgen deprivation therapy AKT AKT serine/threonine kinase 1 ALK1 activin receptor like kinase 1 AMACR alpha-methylacyl-CoA racemase AR androgen receptor AR-V7 androgen receptor splice variant 7 ASCL1 achaete-scute homolog 1 BAMBI BMP and activin membrane-bound inhibitor BCA bicinchoninic acid BMP bone morphogenetic protein bp base pair BPH benign prostate hyperplasia BSA bovine serum albumin CD24 cluster of differentiation 24 CDH1 E-cadherin CDH2 N-cadherin CDK1 cyclin-dependent kinase 1 cDNA complementary DNA CFU colony forming units CHD1 chromodomain helicase DNA binding protein 1 CHGA and B chromogranin A and B CK cytokeratin CNA Copy number alteration Cp crossing point CRPC castration-resistant prostate cancer CTC circulating tumor cell DHT dihydrotestosterone DMSO dimethyl sulfoxide DNA deoxyribonucleic acid DNase deoxyribunuclease DNMT DNA methyltransferase Dox doxycycline EGFR epidermal growth factor receptor EMT epithelial-to-mesenchymal transition ERG V-ets erythroblastosis virus E26 homolog (avian) ERK2 extracellular signal-regulated kinases, alias of MAPK1 Ev empty vector 6 FC fold change FN1 fibronectin 1 FPKM fragments per kilobase of exon per million fragments mapped FZD4 frizzled 4 GAPDH glyceraldehyde-3-phosphate dehydrogenase GO gene ontology HGPIN high-grade prostatic intraepithelial neoplasia HLA-DMB major histocompatibility complex, class II, DM beta ICGC-EOPC International Cancer Genome Consortium-Early Onset PCa project ID1 and 2 inhibitor of differentiation (1 and 2) IGP NGFN IG Prostate Cancer project INSM1 insulinoma associated-1 IPA Ingenuity Pathway Analysis JNK c-Jun N-terminal kinases L1CAM L1 cell adhesion molecule LRP5 and 6 low-density lipoprotein receptor-related protein 5 and 6 MAPK mitogen-activated protein kinase MET mesenchymal-to-epithelial transition miR micro-RNA MMP matrix metalloproteinase MSMB microseminoprotein beta MYCN N-myc proto-oncogene NCAM neuronal cell adhesion molecule NED neuroendocrine differentiation NEPC neuroendocrine prostate cancer NSE neuron specific enolase ORF open reading frame p38 alias of MAPK14, mitogen-activated protein kinase 14 pAKT phospho-AKT PAP prostatic acid phosphatase PCa prostate cancer PI3K phosphatidylinositol 3-kinase PIN prostatic intraepithelial neoplasia PLA1A phospholipase A1 member A PLAT plasminogen activator, tissue type p-p38 phospho-p38 PSA prostate specific antigen pSMAD phospho-SMAD qPCR quantitative reverse transcription PCR RELN reelin 7 REST RE-1 silencing transcription factor/neuron-restrictive silencer factor (NRSF) rhALK1 recombinant decoy receptor ALK1 rhFZD4 recombinant decoy receptor FZD4 RPKM reads per kilobase per million mapped reads RT-PCR reverse transcription PCR SCG3 secretogranin III siRNA small interfering RNA SLC45A3 solute carrier family 45 member 3 SMAD SMAD family protein SNAI2 snail family transcriptional repressor 2 SYP synaptophysin TCF7L2 T cell factor/lymphoid enhancer 2 TCF/LEF-1 transcription factor/lymphoid enhancer binding factor 1 TCGA The Cancer Genome Atlas TDRD1 tudor domain containing 1 T/E TMPRSS2:ERG TGF-β transforming growth factor beta TGFB1 and 2 TGF-β 1 and 2 TMPRSS2 transmembrane protease, serine 2 TUBB3 tubulin beta 3 VIM vimentin VTN vitronectin WNT wingless-type family member ZEB1 zinc finger E-box binding homeobox 1 8 General introduction 9 THE EPIDEMIOLOGY OF PROSTATE CANCER Prostate cancer (PCa) is the most prevalent non-cutaneous malignancy accounting for 15% of the cancers diagnosed in men [1]. It is the second leading cause of cancer- related death in men in Western countries with 417,000 new cases and 92,000 deaths in 2012 in Europe [1, 2]. Only few risk determinants for the development of PCa have been defined, such as increasing age, ethnic or geographical origin, and family history, while the influence of lifestyle factors is less well-established [3-5]. Increasing age is by far the most important risk factor. Only 30% of all cases are diagnosed under the age of 65 years in the US [6]. The 10-years risk to develop PCa for men aged 30 years is 0.01%, increases to 4.77% for men aged 60 years and to 5.50% for men aged 70 years [6]. In 2014, the highest incidence rates for PCa in the US were in the age group of 65-74 years, while between 1975-1995 the incidence rates were in the group of 75 years or older indicating that the diagnosis is currently made at younger age [7]. The incidence rates for PCa are highly variable between different geographical areas, with highest incidence in industrialized countries [6]. Since autopsy-based detection of PCa revealed similar prevalence rates among different geographical areas, the increasing incidence is significantly associated with the widespread implementation of PSA testing leading to elevated biopsy-based detection rates in asymptomatic patients [8, 9]. The mortality pattern has largely remained unchanged and shows less variation worldwide, as PSA testing has a greater impact on incidence than on mortality [6, 8]. Family history is an important contributing factor for the incidence of PCa (men with affected
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