Abstract
Testicular germ-cell tumours of young adults are derived from a pre-invasive intratubular lesion, carcinoma in situ (CIS). In a recent genome-wide gene expression screening using cDNA microarrays, we found PDPN over-expressed in CIS compared to normal adult testis. PDPN encodes podoplanin (Aggrus, human gp36, T1A-2), a transmembrane glycoprotein expressed in lymphatic endothelium and various solid tumours. To examine a potential role for PDPN in testicular neoplasms and during testicular development, we investigated its expression pattern during the development of human testis and in a series of testicular CIS, gonadoblastoma and overt germ-cell tumours. We established by RT-PCR and by immunohistochemistry with a gp36 antibody that PDPN mRNA and the protein product were expressed in testes with germ-cell neoplasms but not in the normal adult testis. We also found gp36 expression in early foetal gonocytes and immature Sertoli cells, similar to the expression pattern of M2A antigen, a previously identified marker for CIS and seminoma. This reinforced our previous proposal that M2A (D2-40) antigen was identical to gp36 (podoplanin, Aggrus, T1A-2). Our findings also suggest that podoplanin has a function in developing testis, most likely at the level of cell–cell interactions among pre-meiotic germ cells and immature Sertoli cells.
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Almstrup K, Hoei-Hansen CE, Wirkner U, Blake J, Schwager C, Ansorge W, Nielsen JE, Skakkebaek NE, Rajpert-De Meyts E, Leffers H (2004) Embryonic stem cell-like features of testicular carcinoma in situ revealed by genome-wide gene expression profiling. Cancer Res 64:4736–4743
Arigami T, Natsugoe S, Uenosono Y, Arima H, Mataki Y, Ehi K, Yanagida S, Ishigami S, Hokita S, Aikou T (2005) Lymphatic invasion using D2-40 monoclonal antibody and its relationship to lymph node micrometastasis in pN0 gastric cancer. Br J Cancer 93:688–693
Avivi-Green C, Kahn HJ, Zimmer G, Sutherland R, Letarte M, Law J, Chen L, Herrler G, Marks A (2004) D2-40 antigen and endoglin are complementary markers of lymphatic and blood capillaries, respectively, in breast cancer. Proceedings of the 95th annual meeting of the American Association for Cancer Research, vol 45, abstract 3243
Bailey D, Baumal R, Law J, Sheldon K, Kannampuzha P, Stratis M, Kahn H, Marks A (1986) Production of a monoclonal antibody specific for seminomas and dysgerminomas. Proc Natl Acad Sci U S A 83:5291–5295
Baumal R, Bailey D, Giwercman A, Skakkebaek N, Stratis M, Marks A (1989) A novel maturation marker for human Sertoli cells. Int J Androl 12:354–359
Breiteneder-Geleff S, Matsui K, Soleiman A, Meraner P, Poczewski H, Kalt R, Schaffner G, Kerjaschki D (1997) Podoplanin, novel 43-kd membrane protein of glomerular epithelial cells, is down-regulated in puromycin nephrosis. Am J Pathol 151:1141–1152
Breiteneder-Geleff S, Soleiman A, Kowalski H, Horvat R, Amann G, Kriehuber E, Diem K, Weninger W, Tschachler E, Alitalo K, Kerjaschki D (1999) Angiosarcomas express mixed endothelial phenotypes of blood and lymphatic capillaries: podoplanin as a specific marker for lymphatic endothelium. Am J Pathol 154:385–394
Franke FE, Pauls K, Rey R, Marks A, Bergmann M, Steger K (2004) Differentiation markers of Sertoli cells and germ cells in fetal and early postnatal human testis. Anat Embryol (Berl) 209:169–177
Fukunaga M (2005) Expression of D2-40 in lymphatic endothelium of normal tissues and in vascular tumours. Histopathology 46:396–402
Giwercman A, Cantell L, Marks A (1991) Placental-like alkaline phosphatase as a marker of carcinoma-in-situ of the testis. Comparison with monoclonal antibodies M2A and 43-9F. APMIS 99:586–594
Giwercman A, Marks A, Bailey D, Baumal R, Skakkebaek NE (1988) A monoclonal antibody as a marker for carcinoma in situ germ cells of the human adult testis. APMIS 96:667–670
Giwercman A, von der MH, Skakkebaek NE (1993) Epidemiological and clinical aspects of carcinoma in situ of the testis. Eur Urol 23:104–110
Hoei-Hansen CE, Almstrup K, Nielsen JE, Brask SS, Graem N, Skakkebaek NE, Leffers H, Rajpert-De Meyts E (2005) Stem cell pluripotency factor NANOG is expressed in human fetal gonocytes, testicular carcinoma in situ and germ cell tumours. Histopathology 47:48–56
Honecker F, Kersemaekers AM, Molier M, Van Weeren PC, Stoop H, De Krijger RR, Wolffenbuttel KP, Oosterhuis W, Bokemeyer C, Looijenga LH (2004) Involvement of E-cadherin and beta-catenin in germ cell tumours and in normal male fetal germ cell development. J Pathol 204:167–174
Jorgensen N, Giwercman A, Muller J, Skakkebaek NE (1993) Immunohistochemical markers of carcinoma in situ of the testis also expressed in normal infantile germ cells. Histopathology 22:373–378
Jorgensen N, Muller J, Jaubert F, Clausen OP, Skakkebaek NE (1997) Heterogeneity of gonadoblastoma germ cells: similarities with immature germ cells, spermatogonia and testicular carcinoma in situ cells. Histopathology 30:177–186
Jorgensen N, Rajpert-De Meyts E, Graem N, Muller J, Giwercman A, Skakkebaek NE (1995) Expression of immunohistochemical markers for testicular carcinoma in situ by normal human fetal germ cells. Lab Invest 72:223–231
Kahn HJ, Bailey D, Marks A (2002) Monoclonal antibody D2-40, a new marker of lymphatic endothelium, reacts with Kaposi’s sarcoma and a subset of angiosarcomas. Mod Pathol 15:434–440
Kahn HJ, Marks A (2002) A new monoclonal antibody, D2-40, for detection of lymphatic invasion in primary tumors. Lab Invest 82:1255–1257
Kato Y, Fujita N, Kunita A, Sato S, Kaneko M, Osawa M, Tsuruo T (2003) Molecular identification of Aggrus/T1alpha as a platelet aggregation-inducing factor expressed in colorectal tumors. J Biol Chem 278:51599–51605
Kato Y, Sasagawa I, Kaneko M, Osawa M, Fujita N, Tsuruo T (2004) Aggrus: a diagnostic marker that distinguishes seminoma from embryonal carcinoma in testicular germ cell tumors. Oncogene 23:8552–8556
Marks A, Sutherland DR, Bailey D, Iglesias J, Law J, Lei M, Yeger H, Banerjee D, Baumal R (1999) Characterization and distribution of an oncofetal antigen (M2A antigen) expressed on testicular germ cell tumours. Br J Cancer 80:569–578
Matsui K, Breiteneder-Geleff S, Kerjaschki D (1998) Epitope-specific antibodies to the 43-kD glomerular membrane protein podoplanin cause proteinuria and rapid flattening of podocytes. J Am Soc Nephrol 9:2013–2026
Meng FJ, Zhou Y, Skakkebaek NE, Marks A, Giwercman A (1996) Detection and enrichment of carcinoma-in-situ cells in semen by an immunomagnetic method using monoclonal antibody M2A. Int J Androl 19:365–370
Niakosari F, Kahn HJ, Marks A, From L (2005) Detection of lymphatic invasion in primary melanoma with monoclonal antibody D2-40: a new selective immunohistochemical marker of lymphatic endothelium. Arch Dermatol 141:440–444
Ordonez NG (2005) Immunohistochemical diagnosis of epithelioid mesothelioma: an update. Arch Pathol Lab Med 129:1407–1414
Rajpert-De Meyts E, Bartkova J, Samson M, Hoei-Hansen CE, Frydelund-Larsen L, Bartek J, Skakkebaek NE (2003) The emerging phenotype of the testicular carcinoma in situ germ cell. APMIS 111:267–278
Rajpert-De Meyts E, Hanstein R, Jorgensen N, Graem N, Vogt PH, Skakkebaek NE (2004) Developmental expression of POU5F1 (OCT-3/4) in normal and dysgenetic human gonads. Hum Reprod 19:1338–1344
Rajpert-De Meyts E, Jorgensen N, Graem N, Muller J, Cate RL, Skakkebaek NE (1999) Expression of anti-Mullerian hormone during normal and pathological gonadal development: association with differentiation of Sertoli and granulosa cells. J Clin Endocrinol Metab 84:3836–3844
Ramirez MI, Millien G, Hinds A, Cao Y, Seldin DC, Williams MC (2003) T1alpha, a lung type I cell differentiation gene, is required for normal lung cell proliferation and alveolus formation at birth. Dev Biol 256:61–72
Schacht V, Dadras SS, Johnson LA, Jackson DG, Hong YK, Detmar M (2005) Up-regulation of the lymphatic marker podoplanin, a mucin-type transmembrane glycoprotein, in human squamous cell carcinomas and germ cell tumors. Am J Pathol 166:913–921
Schacht V, Ramirez MI, Hong YK, Hirakawa S, Feng D, Harvey N, Williams M, Dvorak AM, Dvorak HF, Oliver G, Detmar M (2003) T1alpha/podoplanin deficiency disrupts normal lymphatic vasculature formation and causes lymphedema. EMBO J 22:3546–3556
Skakkebaek NE (1972) Possible carcinoma-in-situ of the testis. Lancet 2:516–517
Skakkebaek NE, Berthelsen JG, Giwercman A, Muller J (1987) Carcinoma-in-situ of the testis: possible origin from gonocytes and precursor of all types of germ cell tumours except spermatocytoma. Int J Androl 10:19–28
Zimmer G, Oeffner F, Von M, V, Tschernig T, Groness HJ, Klenk HD, Herrler G (1999) Cloning and characterization of gp36, a human mucin-type glycoprotein preferentially expressed in vascular endothelium. Biochem J 341(Pt 2):277–284
Acknowledgements
The authors wish to thank L. Andersen and H. Kistrup for the skilful technical assistance, Drs. N. Jørgensen and N. Græm for providing some foetal and prepubertal testis samples, and Dr. P. Andrews for kindly providing TRA-1-60 antibody. This work was supported by grants from the Danish Cancer Society, Svend Andersen’s Foundation, the Danish Medical Research Council, the Kirsten and Freddy Johansen’s Foundation, Willumsen’s Foundation and the Vissing Foundation. The experiments presented in the present manuscript comply with the Danish law.
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Sonne, S.B., Herlihy, A.S., Hoei-Hansen, C.E. et al. Identity of M2A (D2-40) antigen and gp36 (Aggrus, T1A-2, podoplanin) in human developing testis, testicular carcinoma in situ and germ-cell tumours. Virchows Arch 449, 200–206 (2006). https://doi.org/10.1007/s00428-006-0223-4
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DOI: https://doi.org/10.1007/s00428-006-0223-4