Sub theme 3.1.3
Bladder cancer

Goals of research: general outline
Scientific achievements
Future plans: special goals and approach
Running projects
Associated staff

Goals of research: general outline

Research on bladder cancer concerns two aspects. First we are interested the genes that play a role in development of the different subtypes of bladder and upper urinary tract urothelial tumors. We aim to establish how these genetic and epigenetic modifications play a role in recurrence rate, progression and disease course and in addition whether they can be used for stratification of patients for follow-up and/or therapy. Our second goal is to develop simple diagnostic assays that are able to detect recurrent tumors in voided urine in order to reduce the cystoscopy frequency during surveillance as well as urine-based assays that can be of use for early detection of primary urothelial cancer.

Scientific achievements

We have shown that mutations in the FGFR3 oncogene and the TP53 tumor suppressor gene are mutually exclusive and associated with different bladder cancer subtypes with divergent clinical behavior. Stage Ta tumors can further be divided in a group with FGFR3 mutations with a stable genome and a group that lacks mutations in FGFR3 and is often genomically instable. We showed in many publications that FGFR3 mutations are associated with a favorable prognosis. We calculate that in the Netherlands with a population of 16 million 60,000 cystoscopies are being carried out per year to detect recurrent disease in patients with non-muscle invasive bladder cancer (80% of all cases). We have shown that molecular diagnosis of cancer cells in urine by microsatellite- and FGFR3 mutation analysis enables a highly sensitive detection of urothelial cell carcinoma in a multicenter randomized trial. In addition, we observed that informing the urologist of a positive urine test leads to a higher number of detected recurrences and the recurrences are found earlier. Germline mutations in FGFR3 cause autosomal dominant skeletal disorders, sometimes with cutaneous features resembling epidermal nevi. Epidermal nevi are also common congenital skin lesions with an incidence of 1 in 1,000 people. We screened epidermal nevi of this type for FGFR3 mutations. Our results showed that a large proportion of epidermal nevi are caused by mosaicism of FGFR3 mutations in the epidermis, secondary to a postzygotic mutation in early embryonic development.

Future plans: special goals and approach

Genes on chromosome 9 are important in bladder cancers. To establish which genes are involved, we have designed a custom CGH array with probes for all 1400 genes. DNA from 150 tumors was hybridized to the array. Candidate genes that were detected will be assayed for mutations in a second cohort. We have further used a genome wide (Agilent 244 K array) screen for CpG island methylation in 45 bladder tumors of different stages. The identified CpGs were validated in a custom Illumina array. The most significant probes were patented. Future work will be directed at differences in methylation and hence pathogenesis between subgroups, establishing possible prognostic use and the use of probes methylated in all bladder tumors for urine diagnostics. We are involved in a EU FP7 project entitled “Prediction of bladder cancer disease course using risk scores that combine molecular and clinical risk factors”. In this project we lead a workpackage in which early detection of recurrent bladder cancer by urine assays (microsatellite assay, FGFR3, methylation) is investigated in order to reduce the number of cystoscopies for follow-up of these patients. Both a retrospective and a prospective study will be performed. With the final results we will develop a decision model in which patients are stratified based on the genomic alterations in their incident tumor and that will advise on which urine test to use and on the relative frequency of both urine tests and cystoscopies for surveillance of each group of patients.

Most recent publications
  1. B.W.G. van Rhijn, Th.H. van der Kwast, A.N. Vis, W.J. Kirkels, E.R. Boeve, A.C. Jöbsis and E.C. Zwarthoff, FGFR3 and P53 characterize alternative pathways in the pathogenesis of urothelial cell carcinoma, Cancer Res 2004, 64:1911-4.
  2. Magda A. Meester-Smoor, Marcel Vermeij, Marjolein J.L. van Helmond, Anco C. Molijn, Karel H.M. van Wely, Arnold C.P. Hekman, Christl Vermey-Keers, Peter H.J. Riegman, Ellen C. Zwarthoff,Targeted disruption of the Mn1 oncogene results in severe defects in development of membranous bones of the cranial skeleton. Mol Cell. Biol., 25 (2005) 4229-4236.
  3. Johanna M.M. van Oers, I. Lurkin, A.J.A. van Exsel, Y. Nijsen, B.W.G. van Rhijn, Madelon N.M. van der Aa, Ellen C. Zwarthoff, A simple and fast method for the simultaneous detection of 9 FGFR3 mutations in bladder cancer and voided urine, Clin. Cancer Res. 11 (2005):7743-8.
  4. Christian Hafner, Johanna M.M. van Oers, Thomas Vogt, Michael Landthaler, Robert Stoehr, Ferdinand Hofstaedter, Ellen C. Zwarthoff, Arndt Hartmann, Mosaicism of activating FGFR3 mutations in human skin causes epidermal nevi, J. Clin Investigation, 116 (2006) 2201-2207.
  5. Karel H.M. van Wely, Magda A. Meester-Smoor, Albert-Jan Aarnoudse and Ellen C. Zwarthoff, MN1-TEL acts as a dominant negative mutant in RAR-RXR mediated transcription, Oncogene, 26 (2007) 5733-5740.
  6. Johanna M. M. van Oers*, Peter J. Wild*, Maximilian Burgerc, Stefan Denzingerc, Robert Stoehrc, Elke Rosskopfd, Ferdinand Hofstaedterd, Ewout W. Steyerberge, Monika Klinkhammer-Schalkef, Ellen C. Zwarthoffa, Theodorus H. van der Kwasta, and Arndt Hartmann, FGFR3 mutations and a normal CK20 staining pattern define low-grade noninvasive urothelial bladder tumours, Eur Urol, 2007 Sep;52(3):760-8.
  7. Maximilian Burger *1 , Madelon NM van der Aa*3, Johanna MM van Oers3, Anke Brinkmann2, Theodorus H van der Kwast3,4, Ewout C Steyerberg5, Robert Stoehr6,Wim J  Kirkels7,Stefan Denzinger1, Peter J Wild8, Wolf F Wieland1 ,Ferdinand Hofstaedter2, Arndt Hartmann#6, Ellen C Zwarthoff#3, Prediction of progression of non-muscle invasive bladder cancer by WHO 1973 and 2004 grading and by FGFR3 mutation status: a prospective study, European Urology 54 (2008) 835-844.
  8. Madelon N.M. van der Aa, Ellen C. Zwarthoff, Ewout W. Steyerberg, Merel W. Boogaard, Yvette Nijsen, Kirsten A van der Keur, Antonius J. A. van Exsel, Wim J. Kirkels, Chris Bangma,Theo H. van der Kwast, Microsatellite analysis of voided urine samples for surveillance of low grade non-muscle invasive urothelial carcinoma: feasibility and clinical utility in a prospective multicenter study (cefub). European Urology 2009 Mar;55(3):659-67.
  9. MA Meester-Smoor, MJFW Janssen, GC Grosveld, A. de Klein, WFK van IJcken, H. Douben, EC Zwarthoff, MN1 affects expression of genes involded in hematopoiesis and can enhance as well as inhibite RAR/RXR induced gene expression, Carcinogenesis 2008 Jul 16.
  10. LC Kompier, MNM van der Aa, I Lurkin, M Vermeij, WJ Kirkels, CH Bangma, ThH van der Kwast and EC Zwarthoff, The development of multiple bladder tumor recurrences in relation to the FGFR3 mutation status of the primary tumor, J Path, 2009 May;218(1):104-12