Advertisement

Immunohistochemical assessment of basal and luminal markers in non-muscle invasive urothelial carcinoma of bladder

  • Maria Del Carmen Rodriguez Pena
  • Alcides Chaux
  • Marie-Lisa Eich
  • Aline C. Tregnago
  • Diana Taheri
  • Walaa Borhan
  • Rajni Sharma
  • M. Katayoon Rezaei
  • George J. NettoEmail author
Original Article

Abstract

The Cancer Genome Atlas project introduced genomic taxonomy of basal and luminal molecular subtypes in muscle invasive bladder cancer. Fewer studies have addressed the molecular classification in non-muscle invasive bladder cancer (NMIBC). Our aim is to assess the applicability of the proposed phenotypic classification for NMIBC. Three TMAs were constructed from 193 TURBT specimens of 60 bladder cancer patients treated at one of the authors’ institutions (1998–2008). Follow-up data on recurrence, grade, or stage progression was obtained. Immunohistochemistry was performed using an automated Ventana System for markers indicative of luminal (GATA3, CK20, ER, Uroplakin II, and HER2/neu) and basal (CK5/6 and CD44) phenotype. Marker expression was evaluated by 3 urologic pathologists. Using unadjusted logistic regression, we found significant association between tumor recurrence at next biopsy and CD44 expression (OR = 2.51, P = 0.03), tumor recurrence at any subsequent biopsy and ER expression (OR = 0.24, P = 0.04), and tumor grade progression at any subsequent biopsy and HER2/neu expression (OR = 0.24, P = 0.04). After adjusting for pathologic stage, we found a significant association between CK5/6 expression and tumor stage progression at either next or any subsequent biopsy (OR = 0.94, P = 0.006; and OR = 0.97, P = 0.02, respectively). Our findings suggest that individual immunohistochemical markers may be of value as prognostic factors in NMIBC.

Keywords

Molecular classification Basal Luminal Immunohistochemistry Urothelial carcinoma Non-muscle invasive bladder cancer 

Notes

Author contributions

GJN conceived and designed the study and wrote, edited, and reviewed the manuscript. MCRP collected the data and wrote, edited, and reviewed the manuscript. AC and MLE analyzed the data and wrote, edited, and reviewed the manuscript. ACT collected the data and reviewed the manuscript. DT and WB collected the data and reviewed the manuscript. MKR collected samples and clinical data and reviewed the manuscript. RS performed the immunohistochemical analysis and reviewed the manuscript.

Funding

This study is supported in part by a grant from The Johns Hopkins Greenberg Bladder Cancer Institute and The Patana Fund of the Brady Urological Institute, Baltimore, MD.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

All authors gave final approval for publication. GJN takes full responsibility for the work as a whole, including the study design, access to data, and the decision to submit and publish the manuscript.

Informed consent

The Institutional Review Board approved this study.

Supplementary material

428_2019_2618_MOESM1_ESM.jpg (381 kb)
Supplementary Fig 1 A-F: Boxplots illustrating the correlation of marker expression and outcome. Marker score is depicted on y axis. Lighter box plots correspond to presence of event (e.g. recurrence or progression of tumor) while darker box plots correspond to absence of event. Labels on top of the boxplots correspond to adjusted (Bonferroni’s method) p values using Mann-Whitney’s U test (JPG 380 kb)
428_2019_2618_MOESM2_ESM.jpg (107 kb)
Supplementary Fig 2 A-F: Association of outcome measures with basal vs luminal dichotomous categorization of each tumor using CD44 and CK5/6 scores as surrogate for basal category and CK20 and Uroplakin II for luminal category. The highest score of any of the above 4 surrogate markers was applied to assign a basal vs luminal category in a given tumor (JPG 107 kb)
428_2019_2618_MOESM3_ESM.jpg (108 kb)
Supplementary Fig 3 A-F: Association of outcome measures with basal vs luminal dichotomous categorization of each tumor using CD44 and CK5/6 scores as surrogate for basal category and CK20 and Uroplakin II for luminal category. The higher value among the sum of the scores of the two markers in each category was applied to assign a basal vs luminal category in a given tumor (JPG 107 kb)
428_2019_2618_MOESM4_ESM.docx (14 kb)
ESM 4 (DOCX 14 kb)
428_2019_2618_MOESM5_ESM.docx (14 kb)
ESM 5 (DOCX 14 kb)

References

  1. 1.
    Siegel RL, Miller KD, Jemal A (2019) Cancer statistics, 2019. CA Cancer J Clin 69:7–34.  https://doi.org/10.3322/caac.21551 CrossRefPubMedGoogle Scholar
  2. 2.
    Babjuk M, Bohle A, Burger M, Capoun O, Cohen D, Comperat EM, Hernandez V, Kaasinen E, Palou J, Roupret M, van Rhijn BW, Shariat SF, Soukup V, Sylvester RJ, Zigeuner R (2017) EAU guidelines on non-muscle-invasive urothelial carcinoma of the bladder: update 2016. Eur Urol 71:447–461.  https://doi.org/10.1016/j.eururo.2016.05.041 CrossRefPubMedGoogle Scholar
  3. 3.
    Rosenberg JE, Hoffman-Censits J, Powles T, van der Heijden MS, Balar AV, Necchi A, Dawson N, O’Donnell PH, Balmanoukian A, Loriot Y, Srinivas S, Retz MM, Grivas P, Joseph RW, Galsky MD, Fleming MT, Petrylak DP, Perez-Gracia JL, Burris HA, Castellano D, Canil C, Bellmunt J, Bajorin D, Nickles D, Bourgon R, Frampton GM, Cui N, Mariathasan S, Abidoye O, Fine GD, Dreicer R (2016) Atezolizumab in patients with locally advanced and metastatic urothelial carcinoma who have progressed following treatment with platinum-based chemotherapy: a single-arm, multicentre, phase 2 trial. Lancet 387:1909–1920.  https://doi.org/10.1016/s0140-6736(16)00561-4 CrossRefPubMedGoogle Scholar
  4. 4.
    Sharma P, Retz M, Siefker-Radtke A, Baron A, Necchi A, Bedke J, Plimack ER, Vaena D, Grimm MO, Bracarda S, Arranz JA, Pal S, Ohyama C, Saci A, Qu X, Lambert A, Krishnan S, Azrilevich A, Galsky MD (2017) Nivolumab in metastatic urothelial carcinoma after platinum therapy (CheckMate 275): a multicentre, single-arm, phase 2 trial. Lancet Oncol 18:312–322.  https://doi.org/10.1016/s1470-2045(17)30065-7 CrossRefPubMedGoogle Scholar
  5. 5.
    Sjodahl G, Lovgren K, Lauss M, Patschan O, Gudjonsson S, Chebil G, Aine M, Eriksson P, Mansson W, Lindgren D, Ferno M, Liedberg F, Hoglund M (2013) Toward a molecular pathologic classification of urothelial carcinoma. Am J Pathol 183:681–691.  https://doi.org/10.1016/j.ajpath.2013.05.013 CrossRefPubMedGoogle Scholar
  6. 6.
    Cancer Genome Atlas Research N (2014) Comprehensive molecular characterization of urothelial bladder carcinoma. Nature 507:315–322.  https://doi.org/10.1038/nature12965 CrossRefGoogle Scholar
  7. 7.
    Inamura K (2018) Bladder cancer: new insights into its molecular pathology. Cancers 10:100.  https://doi.org/10.3390/cancers10040100 CrossRefGoogle Scholar
  8. 8.
    McConkey DJ, Choi W, Dinney CP (2015) Genetic subtypes of invasive bladder cancer. Curr Opin Urol 25:449–458.  https://doi.org/10.1097/MOU.0000000000000200 CrossRefPubMedGoogle Scholar
  9. 9.
    Choi W, Porten S, Kim S, Willis D, Plimack ER, Hoffman-Censits J, Roth B, Cheng T, Tran M, Lee IL, Melquist J, Bondaruk J, Majewski T, Zhang S, Pretzsch S, Baggerly K, Siefker-Radtke A, Czerniak B, Dinney CP, McConkey DJ (2014) Identification of distinct basal and luminal subtypes of muscle-invasive bladder cancer with different sensitivities to frontline chemotherapy. Cancer Cell 25:152–165.  https://doi.org/10.1016/j.ccr.2014.01.009 CrossRefPubMedGoogle Scholar
  10. 10.
    Robertson AG, Kim J, H A-A, J B, G G, AD C, T H, PW L, KA H, R A, MAA C, EA G, RS K, DA G, SA S, F S-V, DE H, BA C, VE R, X S, B d SC, VS C, KL M, S S, CS P, Y L, LJ K, J Z, C C, AI O, S B, KM L, TM L, CJ W, N S, G G, M M, GB M, DJ MC, Research Network TCGA, JN W, DJ K, SP L (2017) Comprehensive molecular characterization of muscle-invasive bladder cancer. Cell 171:540–556.e525.  https://doi.org/10.1016/j.cell.2017.09.007 CrossRefPubMedGoogle Scholar
  11. 11.
    Hedegaard J, Lamy P, Nordentoft I, Algaba F, Hoyer S, Ulhoi BP, Vang S, Reinert T, Hermann GG, Mogensen K, Thomsen MBH, Nielsen MM, Marquez M, Segersten U, Aine M, Hoglund M, Birkenkamp-Demtroder K, Fristrup N, Borre M, Hartmann A, Stohr R, Wach S, Keck B, Seitz AK, Nawroth R, Maurer T, Tulic C, Simic T, Junker K, Horstmann M, Harving N, Petersen AC, Calle ML, Steyerberg EW, Beukers W, van Kessel KEM, Jensen JB, Pedersen JS, Malmstrom PU, Malats N, Real FX, Zwarthoff EC, Orntoft TF, Dyrskjot L (2016) Comprehensive transcriptional analysis of early-stage urothelial carcinoma cancer cell. 30:27–42.  https://doi.org/10.1016/j.ccell.2016.05.004
  12. 12.
    Breyer J, Wirtz RM, Otto W, Erben P, Kriegmair MC, Stoehr R, Eckstein M, Eidt S, Denzinger S, Burger M, Hartmann A, Consortium B (2017) In stage pT1 non-muscle-invasive bladder cancer (NMIBC), high KRT20 and low KRT5 mRNA expression identify the luminal subtype and predict recurrence and survival. Virchows Arch 470:267–274.  https://doi.org/10.1007/s00428-017-2064-8 CrossRefPubMedGoogle Scholar
  13. 13.
    Wolff AC, Hammond ME, Hicks DG, Dowsett M, McShane LM, Allison KH, Allred DC, Bartlett JM, Bilous M, Fitzgibbons P, Hanna W, Jenkins RB, Mangu PB, Paik S, Perez EA, Press MF, Spears PA, Vance GH, Viale G, Hayes DF (2013) Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 31:3997–4013.  https://doi.org/10.1200/jco.2013.50.9984 CrossRefGoogle Scholar
  14. 14.
    Abrahao-Machado LF, Scapulatempo-Neto C (2016) HER2 testing in gastric cancer: an update. World J Gastroenterol 22:4619–4625.  https://doi.org/10.3748/wjg.v22.i19.4619 CrossRefPubMedGoogle Scholar
  15. 15.
    Raspollini MR, Luque RJ, Menendez CL, Bollito E, Brunelli M, Martignoni G, Montironi R, Cheng L, Blanca A, Baroni G, Minervini A, Lopez-Beltran A (2016) T1 high-grade bladder carcinoma outcome: the role of p16, topoisomerase-IIalpha, survivin, and E-cadherin. Hum Pathol 57:78–84.  https://doi.org/10.1016/j.humpath.2016.06.022 CrossRefPubMedGoogle Scholar
  16. 16.
    Warrick JI, Sjodahl G, Kaag M, Raman JD, Merrill S, Shuman L, Chen G, Walter V, DeGraff DJ (2019) Intratumoral heterogeneity of bladder cancer by molecular subtypes and histologic variants. Eur Urol 75:18–22.  https://doi.org/10.1016/j.eururo.2018.09.003 CrossRefPubMedGoogle Scholar
  17. 17.
    Jawhar NM (2009) Tissue microarray: a rapidly evolving diagnostic and research tool. Ann Saudi Med 29:123–127CrossRefPubMedGoogle Scholar
  18. 18.
    Jakobsson L, Chebil G, Marzouka NA, Liedberg F, Sjodahl G (2018) Low frequency of intratumor heterogeneity in bladder cancer tissue microarrays. Bladder Cancer 4:327–337.  https://doi.org/10.3233/blc-180176 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Maria Del Carmen Rodriguez Pena
    • 1
  • Alcides Chaux
    • 2
  • Marie-Lisa Eich
    • 1
  • Aline C. Tregnago
    • 3
  • Diana Taheri
    • 4
  • Walaa Borhan
    • 5
  • Rajni Sharma
    • 3
  • M. Katayoon Rezaei
    • 6
  • George J. Netto
    • 1
    • 3
    Email author
  1. 1.Department of PathologyThe University of Alabama at BirminghamBirminghamUSA
  2. 2.Department of Scientific Research, School of Postgraduate StudiesNorte UniversityAsuncionParaguay
  3. 3.Department of PathologyJohns Hopkins UniversityBaltimoreUSA
  4. 4.Department of Pathology, Isfahan Kidney Diseases Research CenterIsfahan University of Medical SciencesIsfahanIran
  5. 5.Department of Pathology, College of MedicineTaibah UniversityMadinahSaudi Arabia
  6. 6.Department of PathologyGeorge Washington UniversityWashingtonUSA

Personalised recommendations