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TAp63 and ΔNp63 (p40) in prostate adenocarcinomas: ΔNp63 associates with a basal-like cancer stem cell population but not with metastasis

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Abstract

Like other malignancies, prostate tumors are thought to contain cancer stem-like cells (CSCs) that are responsible for growth, metastasis, and therapy resistance. ΔNp63 (also called p40) is a regulator of normal prostate stem/progenitor cell activities and a marker of normal basal epithelial cells. The levels of ΔNp63 are reduced in prostate adenocarcinomas, although there is also evidence that ΔNp63 is involved in CSC regulation and drives metastasis to the bone. We studied metastatic deposits of prostate cancers with isoform-specific ΔNp63 and TAp63 antibodies. We identified p63-positive cells in only 3 of 42 metastatic prostate tumors (7%), including 2/38 (5.3%) “usual-type” adenocarcinomas. ΔNp63 and TAp63 isoforms were present in the nuclei of a small subpopulation (< 1%) of tumor cells in these metastases. ΔNp63-positive cells showed a basal-like cell phenotype (cytokeratin 8- and androgen receptor-negative, high molecular weight cytokeratin- and cytokeratin 19-positive), distinct from the tumor bulk. TAp63-positive cells were similar but were sometimes cytokeratin 8-positive. A subset of ΔNp63-positive tumor cells were CD44-positive, a marker of “basal” CSCs but were not positive for the “epithelial” CSC marker ALDH1. TAp63 was not associated with either of these CSC markers. None of the tumors containing p63-positive cells showed evidence of bone metastasis, compared with 28% of the p63-negative tumors. These data show that both ΔNp63 and TAp63 are present in only a small proportion of prostate adenocarcinomas and do not associate with metastasis. The data suggest heterogeneity of CSCs in prostate cancer, similar to other cancer types.

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References

  1. Shen MM, Rubin MA (2019) Prostate cancer research at the crossroads. Cold Spring Harb Perspect Med 9:a036277. https://doi.org/10.1101/cshperspect.a036277

    Article  CAS  PubMed  Google Scholar 

  2. Wang G, Zhao D, Spring DJ, DePinho RA (2018) Genetics and biology of prostate cancer. Genes Dev 32:1105–1140. https://doi.org/10.1101/gad.315739.118

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Li JJ, Shen MM (2019) Prostate stem cells and cancer stem cells. Cold Spring Harb Perspect Med 9:a030395. https://doi.org/10.1101/cshperspect.a030395

    Article  CAS  PubMed  Google Scholar 

  4. Ali HR, Dawson S-J, Blows FM, Provenzano E, Pharoah PD, Caldas C (2011) Cancer stem cell markers in breast cancer: pathological, clinical and prognostic significance. Breast Cancer Res 13:R118. https://doi.org/10.1186/bcr3061

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Liu Y, Nenutil R, Appleyard MV, Boylan M, Thompson AM, Coates PJ (2014) Lack of correlation of stem cell markers in breast cancer stem cells. Br J Cancer 110:2063–2071. https://doi.org/10.1038/bjc.2014.105

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Hsu C-L, Chung F-H, Chen C-H, Hsu TT, Liu SM, Chung DS, Hsu YF, Chen CL, Ma N, Lee HC (2016) Genotypes of cancer stem cells characterized by epithelial-to-mesenchymal transition and proliferation related functions. Sci Rep 6:32523. https://doi.org/10.1038/srep32523

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Sampayo RG, Bissell MJ (2019) Cancer stem cells in breast and prostate: fact or fiction? Adv Cancer Res 144:315–341. https://doi.org/10.1016/bs.acr.2019.03.010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Crum CP, McKeon FD (2010) p63 in epithelial survival, germ cell surveillance, and neoplasia. Annu Rev Pathol 5:349–371. https://doi.org/10.1146/annurev-pathol-121808-102117

    Article  CAS  PubMed  Google Scholar 

  9. Nekulova M, Holcakova J, Coates P, Vojtesek B (2011) The role of p63 in cancer, stem cells and cancer stem cells. Cell Mol Biol Lett 16:296–327. https://doi.org/10.2478/s11658-011-0009-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Orzol P, Holcakova J, Nekulova M, Nenutil R, Vojtesek B, Coates PJ (2015) The diverse oncogenic and tumour suppressor roles of p63 and p73 in cancer: a review by cancer site. Histol Histopathol 30:503–521. https://doi.org/10.14670/HH-30.503

    Article  CAS  PubMed  Google Scholar 

  11. Melino G, Memmi EM, Pelicci PG, Bernassola F (2015) Maintaining epithelial stemness with p63. Sci Sig 8:re9. https://doi.org/10.1126/scisignal.aaa1033

    Article  CAS  Google Scholar 

  12. Signoretti S, Waltregny D, Dilks J, Isaac B, Lin D, Garraway L, Yang A, Montironi R, McKeon F, Loda M (2000) p63 is a prostate basal cell marker and is required for prostate development. Am J Pathol 157:1769–1775. https://doi.org/10.1016/S0002-9440(10)64814-6

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Signoretti S, Pires MM, Lindauer M, Horner JW, Grisanzio C, Dhar S, Majumder P, McKeon F, Kantoff PW, Sellers WR, Loda M (2005) p63 regulates commitment to the prostate cell lineage. Proc Natl Acad Sci U S A 102:11355–11360. https://doi.org/10.1073/pnas.0500165102

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Pignon J-C, Grisanzio C, Geng Y, Song J, Shivdasani R, Signoretti S (2013) p63-expressing cells are the stem cells of developing prostate, bladder, and colorectal epithelia. Proc Natl Acad Sci U S A 110:8105–8110. https://doi.org/10.1073/pnas.1221216110

    Article  PubMed  PubMed Central  Google Scholar 

  15. Huang Y, Hamana T, Liu J, Wang C, An L, You P, Chang JYF, Xu J, McKeehan WL, Wang F (2015) Prostate sphere-forming stem cells are derived from the P63-expressing basal compartment. J Biol Chem 290:17745–17752. https://doi.org/10.1074/jbc.M115.661033

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Garraway IP, Sun W, Tran CP, Perner S, Zhang B, Goldstein AS, Hahm SA, Haider M, Head CS, Reiter RE, Rubin MA, Witte ON (2010) Human prostate sphere-forming cells represent a subset of basal epithelial cells capable of glandular regeneration in vivo. Prostate 70:491–501. https://doi.org/10.1002/pros.21083

    Article  PubMed  Google Scholar 

  17. Portillo-Lara R, Alvarez MM (2015) Enrichment of the cancer stem phenotype in sphere cultures of prostate cancer cell lines occurs through activation of developmental pathways mediated by the transcriptional regulator ΔNp63α. PLoS One 10:e0130118. https://doi.org/10.1371/journal.pone.0130118

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Nylander K, Vojtesek B, Nenutil R, Lindgren B, Roos G, Zhanxiang W, Sjöström B, Dahlqvist Å, Coates PJ (2002) Differential expression of p63 isoforms in normal tissues and neoplastic cells. J Pathol 198:417–427. https://doi.org/10.1002/path.1231

    Article  CAS  PubMed  Google Scholar 

  19. Sailer V, Stephan C, Wernert N, Perner S, Jung K, Kristiansen G (2013) Comparison of p40 (ΔNp63) and p63 expression in prostate tissues--which one is the superior diagnostic marker for basal cells? Histopathology 63:50–56. https://doi.org/10.1111/his.12116

    Article  PubMed  Google Scholar 

  20. Shah RB, Kunju LP, Shen R, LeBlanc M, Zhou M, Rubin MA (2004) Usefulness of basal cell cocktail (34betaE12 + p63) in the diagnosis of atypical prostate glandular proliferations. Am J Clin Pathol 122:517–523. https://doi.org/10.1309/WRM5-1C70-P1NB-FE4K

    Article  PubMed  Google Scholar 

  21. Ali TZ, Epstein JI (2008) False positive labeling of prostate cancer with high molecular weight cytokeratin: p63 a more specific immunomarker for basal cells. Am J Surg Pathol 32:1890–1895. https://doi.org/10.1097/PAS.0b013e31817ce994

    Article  PubMed  Google Scholar 

  22. Osunkoya AO, Hansel DE, Sun X, Neto GJ, Epstein JI (2008) Aberrant diffuse expression of p63 in adenocarcinoma of the prostate on needle biopsy and radical prostatectomy: report of 21 cases. Am J Surg Pathol 32:461–467. https://doi.org/10.1097/PAS.0b013e318157020e

    Article  PubMed  Google Scholar 

  23. Wu A, Kunju LP (2013) Prostate cancer with aberrant diffuse p63 expression: report of a case and review of the literature and morphologic mimics. Arch Pathol Lab Med 137:1179–1184. https://doi.org/10.5858/arpa.2013-0254-CR

    Article  PubMed  Google Scholar 

  24. Tan H-L, Haffner MC, Esopi DM, Vaghasia AM, Giannico GA, Ross HM, Ghosh S, Hicks JL, Zheng Q, Sangoi AR, Yegnasubramanian S, Osunkoya AO, de Marzo AM, Epstein JI, Lotan TL (2015) Prostate adenocarcinomas aberrantly expressing p63 are molecularly distinct from usual-type prostatic adenocarcinomas. Mod Pathol 28:446–456. https://doi.org/10.1038/modpathol.2014.115

    Article  CAS  PubMed  Google Scholar 

  25. Uchida K, Ross H, Lotan T, Pignon J-C, Signoretti S, Epstein JI, Illei PB (2015) ΔNp63 (p40) expression in prostatic adenocarcinoma with diffuse p63 positivity. Hum Pathol 46:384–389. https://doi.org/10.1016/j.humpath.2014.11.011

    Article  CAS  PubMed  Google Scholar 

  26. Torres A, Alshalalfa M, Davicioni E, Gupta A, Yegnasubramanian S, Wheelan SJ, Epstein JI, de Marzo AM, Lotan TL (2018) ETS2 is a prostate basal cell marker and is highly expressed in prostate cancers aberrantly expressing p63. Prostate 78:896–904. https://doi.org/10.1002/pros.23646

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Rosenbluth JM, Johnson K, Tang L, Triplett, Pientenpol JA (2009) Evaluation of p63 and p73 antibodies for cross-reactivity. Cell Cycle 8:3702–3706. https://doi.org/10.4161/cc.8.22.10036

    Article  CAS  PubMed  Google Scholar 

  28. Nekulova M, Holcakova J, Nenutil R, Stratmann R, Bouchalova P, Müller P, Mouková L, Coates PJ, Vojtesek B (2013) Characterization of specific p63 and p63-N-terminal isoform antibodies and their application for immunohistochemistry. Virchows Arch 463:415–425. https://doi.org/10.1007/s00428-013-1459-4

    Article  CAS  PubMed  Google Scholar 

  29. Lee AHS (2013) Use of immunohistochemistry in the diagnosis of problematic breast lesions. J Clin Pathol 66:471–477. https://doi.org/10.1136/jclinpath-2012-201109

    Article  CAS  PubMed  Google Scholar 

  30. Ribeiro-Silva A, Ramalho LNZ, Garcia SB, Brandao DF, Chahud F, Zucoloto S (2005) p63 correlates with both BRCA1 and cytokeratin 5 in invasive breast carcinomas: further evidence for the pathogenesis of the basal phenotype of breast cancer. Histopathology 47:458–466. https://doi.org/10.1111/j.1365-2559.2005.02249.x

    Article  CAS  PubMed  Google Scholar 

  31. Liu Y, Nekulova M, Nenutil R, Horakova I, Appleyard MV, Murray K, Holcakova J, Galoczova M, Quinlan P, Jordan LB, Purdie CA, Vojtesek B, Thompson AM, Coates PJ (2020) ∆Np63/p40 correlates with the location and phenotype of basal/mesenchymal cancer stem-like cells in human ER+ and HER2+ breast cancers. J Pathol Clin Res 6:83–93. https://doi.org/10.1002/cjp2.149

    Article  CAS  PubMed  Google Scholar 

  32. Kim J, Villadsen R, Sørlie T et al (2012) Tumor initiating but differentiated luminal-like breast cancer cells are highly invasive in the absence of basal-like activity. Proc Natl Acad Sci U S A 109:6124–6129. https://doi.org/10.1073/pnas.1203203109

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Maitland NJ, Collins AT (2008) Prostate cancer stem cells: a new target for therapy. J Clin Oncol 26:2862–2870. https://doi.org/10.1200/JCO.2007.15.1472

    Article  PubMed  Google Scholar 

  34. Parsons JK, Gage WR, Nelson WG, De Marzo AM (2001) p63 protein expression is rare in prostate adenocarcinoma: implications for cancer diagnosis and carcinogenesis. Urology 58:619–624. https://doi.org/10.1016/s0090-4295(01)01311-5

    Article  CAS  PubMed  Google Scholar 

  35. Resetkova E, Reis-Filho JS, Jain RK, Mehta R, Thorat MA, Nakshatri H, Badve S (2010) Prognostic impact of ALDH1 in breast cancer: a story of stem cells and tumor microenvironment. Breast Cancer Res Treat 123:97–108. https://doi.org/10.1007/s10549-009-0619-3

    Article  PubMed  Google Scholar 

  36. Epstein JI, Egevad L, Humphrey PA, Montroni R (2014) Best practices recommendations in the application of immunohistochemistry in the prostate: report from the International Society of Urologic Pathology consensus conference. Am J Surg Pathol 38:e6–e19. https://doi.org/10.1097/PAS.0000000000000238

    Article  PubMed  Google Scholar 

  37. Reiner T, de Las PA, Parrondo R, Perez-Stable C (2007) Progression of prostate cancer from a subset of p63-positive basal epithelial cells in FG/Tag transgenic mice. Mol Cancer Res 5:1171–1179. https://doi.org/10.1158/1541-7786.MCR-07-0024

    Article  CAS  PubMed  Google Scholar 

  38. Di Giacomo V, Tian TV, Mas A et al (2017) ΔNp63α promotes adhesion of metastatic prostate cancer cells to the bone through regulation of CD82. Oncogene 36:4381–4392. https://doi.org/10.1038/onc.2017.42

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Gu G, Yuan J, Wills M, Kasper S (2007) Prostate cancer cells with stem cell characteristics reconstitute the original human tumor in vivo. Cancer Res 67:4807–4815. https://doi.org/10.1158/0008-5472.CAN-06-4608

    Article  CAS  PubMed  Google Scholar 

  40. Kurita T, Medina RT, Mills AA, Cunha GR (2004) Role of p63 and basal cells in the prostate. Development 131:4955–4964. https://doi.org/10.1242/dev.01384

    Article  CAS  PubMed  Google Scholar 

  41. Tucci P, Agostini M, Grespi F, Markert EK, Terrinoni A, Vousden KH, Muller PAJ, Dötsch V, Kehrloesser S, Sayan BS, Giaccone G, Lowe SW, Takahashi N, Vandenabeele P, Knight RA, Levine AJ, Melino G (2012) Loss of p63 and its microRNA-205 target results in enhanced cell migration and metastasis in prostate cancer. Proc Natl Acad Sci U S A 109:15312–15317. https://doi.org/10.1073/pnas.1110977109

    Article  PubMed  PubMed Central  Google Scholar 

  42. Hudson DL, Guy AT, Fry P, O'Hare MJ, Watt FM, Masters JR (2001) Epithelial cell differentiation pathways in the human prostate: identification of intermediate phenotypes by keratin expression. J Histochem Cytochem 49:271–278. https://doi.org/10.1177/002215540104900214

    Article  CAS  PubMed  Google Scholar 

  43. Wang Y, Hayward S, Cao M, Thayer K, Cunha G (2001) Cell differentiation lineage in the prostate. Differentiation 68:270–279

    Article  CAS  Google Scholar 

  44. Liu S, Cong Y, Wang D, Sun Y, Deng L, Liu Y, Martin-Trevino R, Shang L, McDermott SP, Landis MD, Hong S, Adams A, D’Angelo R, Ginestier C, Charafe-Jauffret E, Clouthier SG, Birnbaum D, Wong ST, Zhan M, Chang JC, Wicha MS (2014) Breast cancer stem cells transition between epithelial and mesenchymal states reflective of their normal counterparts. Stem Cell Reports 2:78–91. https://doi.org/10.1016/j.stemcr.2013.11.009

    Article  CAS  PubMed  Google Scholar 

  45. Coates PJ, Nenutil R, Holcakova J, Nekulova M, Podhorec J, Svoboda M, Vojtesek B (2018) p63 isoforms in triple-negative breast cancer: ΔNp63 associates with the basal phenotype whereas TAp63 associates with androgen receptor, lack of BRCA mutation, PTEN and improved survival. Virchows Arch 472:351–359. https://doi.org/10.1007/s00428-018-2324-2

    Article  CAS  PubMed  Google Scholar 

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Funding

This work was supported by the Czech Science Foundation (19-06530S), the European Regional Development Fund - Project ENOCH (No. CZ.02.1.01/0.0/0.0/16_019/0000868) and the Ministry of Health, Czech Republic—conceptual development of research organization (MMCI, 00209805). The MMCI biobank is supported by grant LM2018125 from the Ministry of Education, Youth and Sports and co-funded by ADOPT BBMRI-ERIC, supported by EU Horizon 2020 (grant agreement No. 676550). MG was a Brno Ph.D. Talent Scholarship Holder, funded by the Brno City Municipality.

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  1. Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty kopec 7, 656 53, Brno, Czech Republic

    Michaela Galoczova, Rudolf Nenutil, Zuzana Pokorna, Borivoj Vojtesek & Philip J. Coates

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  1. Michaela Galoczova
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  2. Rudolf Nenutil
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  3. Zuzana Pokorna
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Contributions

MG performed immunohistochemistry and was a major contributor to preparing the manuscript. RN was involved in study concept and design, selected patient material, and interpreted immunohistochemical staining and clinicopathological data. ZP performed immunohistochemistry and assisted with analysis. BV was involved in study design and supervision. PC designed the study, interpreted data, and was a major contributor to writing the manuscript. All authors were involved in revising the manuscript and all authors read and approved the final version.

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Correspondence to Philip J. Coates.

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Conflict of interest

BV is a consultant for and RN is a co-owner of Moravian Biotechnology, who originally produced the p63 monoclonal antibodies used in this study. The company did not provide financial support or have any influence over the design, execution, or interpretation of the data. MG, ZP, and PJC declare that they have no competing interests.

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The study was performed retrospectively on redundant excess tissues. All patient data were anonymized. In accordance with the Declaration of Helsinki and the regulations of the European Union, the use of excess and redundant tissues was approved following local ethical committee review by the Biobanking and Biomolecular resources Research Infrastructure (BBMRI) at the Masaryk Memorial Cancer Institute, Brno. Patient consent is not required for the retrospective use of anonymized, redundant excess tissues.

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Fig. S1

ΔNp63 immunostaining in three lymph node metastases. Sections were stained with immunoperoxidase (brown) and nuclei were counterstained with hematoxylin (blue). Panels on the right show a different field of the same section at higher magnification. (a) Prostate adenocarcinoma sample where only a few neoplastic cells were ΔNp63+. (b) In the other adenocarcinoma sample, ΔNp63+ cells were concentrated in one region of the lymph node tumor deposit, where they represented ~10% of the tumor cells. (c) Carcinosarcoma with ~5% ΔNp63+ tumor cells occurring in one region of the lymph node metastasis. Arrows indicate examples of ∆Np63+ cells. (PNG 1803 kb)

High Resolution Image (TIFF 2505 kb)

Fig. S2

ΔNp63 immunostaining in primary tumor samples in which the corresponding metastasis samples did not contain ΔNp63+ cells. Sections were stained for ΔNp63 with immunoperoxidase (brown) and nuclei were counterstained with hematoxylin. The photomicrographs show four examples of ΔNp63+ cells in radical prostatectomy tissues. In each photomicrograph, whilst the majority of cells are basally located in benign appearing glands, there are occasional cells that are difficult to assign. In all cases, regions such as those shown were found towards the edge of the invasive tumor and were always found together with normal gland structures. (PNG 4493 kb)

High Resolution Image (TIF 5947 kb)

Fig. S3

ΔNp63 co-localization with markers of epithelial differentiation in benign prostate hyperplasia. (a) Dual-peroxidase labelling for ΔNp63 (brown) and the indicated cytokeratins (gray). Nuclei are counterstained with nuclear fast red. The majority of ΔNp63+ cells co-localize with HMWCK and variably with CK19. ΔNp63+ cells are negative for CK8. (b) Immunofluorescence. p63+ cells (red) are negative or weakly positive for AR (green). The merged image shows counterstaining with DAPI (blue). (PNG 1319 kb)

High Resolution Image (TIFF 1875 kb)

Fig. S4

ΔNp63 co-localization with cancer stem cell markers in benign prostate hyperplasia. ΔNp63+ cells (brown) are negative for ALDH1 (gray) and are mostly positive for CD44 (gray). (PNG 770 kb)

High Resolution Image (TIFF 1046 kb)

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Galoczova, M., Nenutil, R., Pokorna, Z. et al. TAp63 and ΔNp63 (p40) in prostate adenocarcinomas: ΔNp63 associates with a basal-like cancer stem cell population but not with metastasis. Virchows Arch 478, 627–636 (2021). https://doi.org/10.1007/s00428-020-02944-z

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