Skip to main content

Advertisement

SpringerLink
Log in

Tumor evolution and progression in multifocal and paired non-invasive/invasive urothelial carcinoma

  • Original Article
  • Published:
Virchows Archiv Aims and scope Submit manuscript

Abstract

Although multifocal tumors and non-invasive/invasive components are commonly encountered in surgical pathology, their genetic relationship is often poorly characterized. We used next-generation sequencing (NGS) to characterize somatic alterations in a patient with five spatially distinct, high-grade papillary urothelial carcinomas (UCs), with one tumor harboring an underlying invasive component. NGS of 409 cancer-related genes was performed on DNA isolated from formalin-fixed paraffin-embedded (FFPE) blocks representing each papillary tumor (n = 5), the invasive component of one tumor, and matched normal tissue. We identified nine unique non-synonymous somatic mutations across the six UC samples, including five present in each carcinoma sample, consistent with clonal origin and limited intertumoral heterogeneity. Copy number and loss of heterogeneity (LOH) profiles were similar in all six carcinomas; however, the invasive carcinoma component uniquely showed focal CDKN2A loss and chromosome 9 LOH and did not harbor gains of chromosomes 5p or X that were present in the other tumor samples. Phylogenetic analysis supported the invasive component arising from a shared progenitor prior to the outgrowth of cells in the non-invasive tumors. Results were extended to three additional cases of upper tract UC with paired non-invasive/invasive components, which identified driving alterations exclusive to both non-invasive and invasive components. Lastly, we performed targeted RNA sequencing (RNAseq) using a custom bladder cancer panel, which confirmed gene expression signature differences between paired non-invasive/invasive components. The results and approaches presented here may be useful in understanding the clonal relationships in multifocal cancers or paired non-invasive/invasive components from routine FFPE specimens.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Babjuk M, Oosterlinck W, Sylvester R et al (2011) EAU guidelines on non-muscle-invasive urothelial carcinoma of the bladder, the 2011 update. Eur Urol 59:997–1008

    Article  PubMed  Google Scholar 

  2. Xylinas E, Rink M, Margulis V et al (2012) Multifocal carcinoma in situ of the upper tract is associated with high risk of bladder cancer recurrence. Eur Urol 61:1069–1070

    Article  PubMed  Google Scholar 

  3. Lin HH, Ke HL, Wu WJ, Lee YH, Chang LL (2012) Hypermethylation of E-cadherin, p16, p14, and RASSF1A genes in pathologically normal urothelium predict bladder recurrence of bladder cancer after transurethral resection. Urol Oncol 30:177–181

    Article  CAS  PubMed  Google Scholar 

  4. Bornman DM, Mathew S, Alsruhe J, Herman JG, Gabrielson E (2001) Methylation of the E-cadherin gene in bladder neoplasia and in normal urothelial epithelium from elderly individuals. Am J Pathol 159:831–835

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Yamamoto S, Romanenko A, Wei M et al (1999) Specific p53 gene mutations in urinary bladder epithelium after the Chernobyl accident. Cancer Res 59:3606–3609

    CAS  PubMed  Google Scholar 

  6. Denzinger S, Mohren K, Knuechel R et al (2006) Improved clonality analysis of multifocal bladder tumors by combination of histopathologic organ mapping, loss of heterozygosity, fluorescence in situ hybridization, and p53 analyses. Hum Pathol 37:143–151

    Article  CAS  PubMed  Google Scholar 

  7. Louhelainen J, Wijkstrom H, Hemminki K (2000) Allelic losses demonstrate monoclonality of multifocal bladder tumors. Int J Cancer 87:522–527

    Article  CAS  PubMed  Google Scholar 

  8. Simon R, Eltze E, Schafer KL et al (2001) Cytogenetic analysis of multifocal bladder cancer supports a monoclonal origin and intraepithelial spread of tumor cells. Cancer Res 61:355–362

    CAS  PubMed  Google Scholar 

  9. Stoehr R, Knuechel R, Boecker J et al (2002) Histologic-genetic mapping by allele-specific PCR reveals intraurothelial spread of p53 mutant tumor clones. Lab Invest 82:1553–1561

    Article  CAS  PubMed  Google Scholar 

  10. Stoehr R, Hartmann A, Hiendlmeyer E, Murle K, Wieland W, Knuechel R (2000) Oligoclonality of early lesions of the urothelium as determined by microdissection-supported genetic analysis. Pathobiology 68:165–172

    Article  CAS  PubMed  Google Scholar 

  11. Cancer Genome Atlas Research Network (2014) Comprehensive molecular characterization of urothelial bladder carcinoma. Nature 507:315–22

  12. Castillo-Martin M, Domingo-Domenech J, Karni-Schmidt O, Matos T, Cordon-Cardo C (2010) Molecular pathways of urothelial development and bladder tumorigenesis. Urol Oncol 28:401–408

    Article  CAS  PubMed  Google Scholar 

  13. Netto GJ (2012) Molecular biomarkers in urothelial carcinoma of the bladder: are we there yet? Nat Rev Urol 9:41–51

    Article  CAS  Google Scholar 

  14. Sidrandsky DF, Frost P, Von Eschenbach A, Oyasu R, Preisinger AC, Vogelstein B (1992) Clonal origin of bladder cancer. N Engl J Med 326:737–740

    Article  Google Scholar 

  15. Gerlinger M, Horswell S, Larkin J et al (2014) Genomic architecture and evolution of clear cell renal cell carcinomas defined by multiregion sequencing. Nat Genet 46:225–23

    Article  CAS  PubMed  Google Scholar 

  16. Gerlinger M, Rowan AJ, Horswell S et al (2012) Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. N Engl J Med 366:883–892

    Article  CAS  PubMed  Google Scholar 

  17. Nordentoft IL P, Birkenkamp-Demtroder K, Shumansky K, Vang S, Hornshoj H, Juul M, Villesen P, Hedegaard J, Roth A, Thorsen K, Hoyer S, Borre M, Reinert T, Fristrup N, Dyrskjot L, Shah S, Pedersen JS, Orntoft TF (2014) Mutational context and diverse clonal developments in early and late bladder cancer. Cell Rep 7:1649–1663

    Article  Google Scholar 

  18. Chang X, Wang K (2012) wANNOVAR: annotating genetic variants for personal genomes via the web. J Med Genet 49:433–436

    Article  PubMed Central  PubMed  Google Scholar 

  19. Wang K, Li M, Hakonarson H (2010) ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. Nucleic Acids Res 38:3

    CAS  Google Scholar 

  20. Grasso C, Butler T, Rhodes K, et al. (2014) Assessing copy number alterations in targeted, amplicon-based next-generation sequencing data. J Mol Diagn. (in press)

  21. Choi W, Porten S, Kim S et al (2014) Identification of distinct basal and luminal subtypes of muscle-invasive bladder cancer with different sensitivities to frontline chemotherapy. Cancer Cell 25:152–165

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Damrauer JS, Hoadley KA, Chism DD et al (2014) Intrinsic subtypes of high-grade bladder cancer reflect the hallmarks of breast cancer biology. Proc Natl Acad Sci U S A 111:3110–3115

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Balbas-Martinez C, Sagrera A, Carrillo-de-Santa-Pau E et al (2013) Recurrent inactivation of STAG2 in bladder cancer is not associated with aneuploidy. Nat Genet 45:1464–1469

    Article  CAS  PubMed  Google Scholar 

  24. Guo G, Sun X, Chen C et al (2013) Whole-genome and whole-exome sequencing of bladder cancer identifies frequent alterations in genes involved in sister chromatid cohesion and segregation. Nat Genet 45:1459–1463

    Article  CAS  PubMed  Google Scholar 

  25. Iyer G, Al-Ahmadie H, Schultz N et al (2013) Prevalence and co-occurrence of actionable genomic alterations in high-grade bladder cancer. J Clin Oncol 31:3133–3140

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  26. Kompier LC, Lurkin I, van der Aa MN, van Rhijn BW, van der Kwast TH, Zwarthoff EC (2010) FGFR3, HRAS, KRAS, NRAS and PIK3CA mutations in bladder cancer and their potential as biomarkers for surveillance and therapy. PLoS One 5:e13821

    Article  PubMed Central  PubMed  Google Scholar 

  27. Rebouissou S, Herault A, Letouze E et al (2012) CDKN2A homozygous deletion is associated with muscle invasion in FGFR3-mutated urothelial bladder carcinoma. J Pathol 227:315–324

  28. Greulich H, Kaplan B, Mertins P et al (2012) Functional analysis of receptor tyrosine kinase mutations in lung cancer identifies oncogenic extracellular domain mutations of ERBB2. Proc Natl Acad Sci U S A 109:14476–14481

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Ross JS, Wang K, Gay LM et al (2014) A high frequency of activating extracellular domain ERBB2 (HER2) mutation in micropapillary urothelial carcinoma. Clin Cancer Res 20:68–75

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank Mandy Davis and Angela Fullen for technical assistance and Arul Chinnaiyan and Eric Fearon for critical review of the manuscript. This work was supported by the University of Michigan, Department of Anatomic Pathology Operations Committee (to JIW and SAT) and generous gifts from the Teeter and MacLeod families for bladder cancer research at the University of Michigan.

Conflict of interest

SAT has a separate sponsored research agreement with Compendia Bioscience/Life Technologies. None of the study described herein was supported by Compendia Bioscience/Life Technologies, and they had no role in the data collection, interpretation, or analysis and did not participate in the study design or the decision to submit for publication. All authors have no disclosures or conflicts of interest otherwise.

Author information

Authors and Affiliations

  1. Michigan Center for Translational Pathology, Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, USA

    Joshua I. Warrick, Anmol Amin, Chia-Jen Liu, Andi K. Cani, Andrew S. McDaniel, Venkata Yadati, Michael J. Quist, Rohit Mehra, Catherine S. Grasso & Scott A. Tomlins

  2. Computational Medicine & Bioinformatics, University of Michigan Medical School, Ann Arbor, MI, USA

    Daniel H. Hovelson

  3. Department of Otolaryngology, University of Michigan Medical School, Ann Arbor, MI, USA

    J. Chad Brenner

  4. Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, MI, USA

    Felix Y. Feng

  5. Department of Urology, University of Michigan Medical School, Ann Arbor, MI, USA

    Alon Z. Weizer, Felix Y. Feng & Scott A. Tomlins

  6. Comprehensive Cancer Center, University of Michigan Medical School, 1524 BSRB, 109 Zina Pitcher Place, Ann Arbor, MI, 48109-2200, USA

    Alon Z. Weizer, Felix Y. Feng & Scott A. Tomlins

  7. Department of Pathology, Oregon Health & Sciences University, Portland, OR, USA

    Michael J. Quist & Catherine S. Grasso

Authors
  1. Joshua I. Warrick
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daniel H. Hovelson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anmol Amin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chia-Jen Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Andi K. Cani
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Andrew S. McDaniel
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Venkata Yadati
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Michael J. Quist
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Alon Z. Weizer
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. J. Chad Brenner
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Felix Y. Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Rohit Mehra
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Catherine S. Grasso
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Scott A. Tomlins
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Scott A. Tomlins.

Additional information

Joshua I. Warrick and Daniel H. Hovelson contributed equally to this manuscript.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 2872 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Warrick, J.I., Hovelson, D.H., Amin, A. et al. Tumor evolution and progression in multifocal and paired non-invasive/invasive urothelial carcinoma. Virchows Arch 466, 297–311 (2015). https://doi.org/10.1007/s00428-014-1699-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00428-014-1699-y

Keywords

Search

Navigation