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Open Access 01-12-2020 | Merkel Cell Carcinoma | Research

Promoter activity of Merkel cell Polyomavirus variants in human dermal fibroblasts and a Merkel cell carcinoma cell line

Authors: Ibrahim Abdulsalam, Kashif Rasheed, Baldur Sveinbjørnsson, Bernhard Ehlers, Ugo Moens

Published in: Virology Journal | Issue 1/2020

Abstract

Background

Merkel cell polyomavirus (MCPyV) is a human polyomavirus that establishes a life-long harmless infection in most individuals, with dermal fibroblasts believed to be the natural host cell. However, this virus is the major cause of Merkel cell carcinoma (MCC), an aggressive skin cancer. Several MCPyV variants with polymorphism in their promoter region have been isolated, but it is not known whether these differences affect the biological properties of the virus.

Methods

Using transient transfection studies in human dermal fibroblasts and the MCC cell line MCC13, we compared the transcription activity of the early and late promoters of the most commonly described non-coding control region MCPyV variant and six other isolates containing specific mutation patterns.

Results

Both the early and late promoters were significantly stronger in human dermal fibroblasts compared with MCC13 cells, and a different promoter strength between the MCPyV variants was observed. The expression of full-length large T-antigen, a viral protein that regulates early and late promoter activity, inhibited early and late promoter activities in both cell lines. Nonetheless, a truncated large T-antigen, which is expressed in virus-positive MCCs, stimulated the activity of its cognate promoter.

Conclusion

The promoter activities of all MCPyV variants tested was stronger in human dermal fibroblasts, a cell line that supports viral replication, than in MCC13 cells, which are not permissive for MCPyV. Truncated large T-antigen, but not full-length large T-antigen stimulated viral promoter activity. Whether, the difference in promoter strength and regulation by large T-antigen may affect the replication and tumorigenic properties of the virus remains to be determined.

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Feng H, Shuda M, Chang Y, Moore PS. Clonal integration of a polyomavirus in human Merkel cell carcinoma. Science. 2008;319:1096–100. https://doi.org/10.1126/science.1152586.CrossRefPubMedPubMedCentral

Kassem A, Schöpflin A, Diaz C, Weyers W, Stickler E, Werner M, Zur HA. Frequent detection of Merkel cell polyomavirus in human Merkel cell carcinomas and identification of a unique deletion in the VP1 gene. Cancer Res. 2008;68:5009–13. https://doi.org/10.1158/0008-5472.CAN-08-0949.CrossRefPubMed

Foulongne V, Kluger N, Dereure O, Brieu N, Guillot B, Segondy M. Merkel cell polyomavirus and Merkel cell carcinoma. France Emerg Dis. 2008;14:1491–3. https://doi.org/10.3201/eid1409.080651.CrossRef

Becker JC, Houben R, Ugurel S, Trefzer U, Pföhler C, Schrama D. MC polyomavirus is frequently present in Merkel cell carcinoma of European patients. J Invest Dermatol. 2009;129:248–50. https://doi.org/10.1038/jid.2008.198.CrossRefPubMed

Van Ghelue M, Moens U. Merkel cell polyomaviurs: a causal factor in Merkel cell carcinoma. In: La Porta CAM, editor. Skin cancers-risk factors, prevention and therapy. Croatiia: IntechOpen; 2011. p. 109–42. https://doi.org/10.5772/25675.CrossRef

Wendzicki JA, Moore PS, Chang Y. Large T and small T antigens of Merkel cell polyomavirus. Curr Opin Virol. 2015;11:38–43. https://doi.org/10.1016/j.coviro.2015.01.009.CrossRefPubMedPubMedCentral

Shuda M, Guastafierro A, Geng X, Shuda Y, Ostrowski SM, Lukianov S, Jenkins FJ, Honda K, Maricich SM, Moore PS, Chang Y. Merkel cell Polyomavirus small T antigen induces Cancer and embryonic Merkel cell proliferation in a transgenic mouse model. PLoS One. e01423292015;10. https://doi.org/10.1371/journal.pone.0142329.

Verhaegen ME, Mangelberger D, Harms PW, Vozheiko TD, Weick JW, Wilbert DM, Saunders TL, Ermilov AN, Bichakjian CK, Johnson TM, Imperiale MJ, Dlugosz AA. Merkel cell polyomavirus small T antigen is oncogenic in transgenic mice. J Invest Dermatol. 2015;135:1415–24. https://doi.org/10.1038/jid.2014.446.CrossRefPubMed

Spurgeon ME, Cheng J, Bronson RT, Lambert PF, DeCaprio, J.A. Tumorigenic activity of merkel cell polyomavirus T antigens expressed in the stratified epithelium of mice. Cancer Res 2015;75:1068–1079. doi: https://doi.org/10.1158/0008-5472.CAN-14-2425.

10.

Bouvard V, Baan RA, Grosse Y, Lauby-Secretan B, El Ghissassi F, Benbrahim-Tallaa L, Guha N, Straif K. Carcinogenicity of malaria and of some polyomaviruses. Lancet Oncol. 2012;13:339–40.CrossRef

11.

Stakaityte G, Wood JJ, Knight LM, Abdul-Sada H, Adzahar NS, Nwogu N, Macdonald A, Whitehouse A. Merkel cell polyomavirus: molecular insights into the most recently discovered human tumour virus. Cancers. 2014;6:1267–97. https://doi.org/10.3390/cancers6031267.CrossRefPubMedPubMedCentral

12.

Borchert S, Czech-Sioli M, Neumann F, Schmidt C, Wimmer P, Dobner T, Grundhoff A, Fischer N. High-affinity Rb binding, p53 inhibition, subcellular localization, and transformation by wild-type or tumor-derived shortened Merkel cell polyomavirus large T antigens. J Virol. 2014;88:3144–60. https://doi.org/10.1128/JVI.02916-13.CrossRefPubMedPubMedCentral

13.

Viscidi RP, Rollison DE, Sondak VK, Silver B, Messina JL, Giuliano AR, Fulp W, Ajidahun A, Rivanera D. Age-specific seroprevalence of Merkel cell polyomavirus, BK virus, and JC virus. Clin Vaccine Immunol. 2011;18:1737–43. https://doi.org/10.1128/CVI.05175-11.CrossRefPubMedPubMedCentral

14.

Kean JM, Rao S, Wang M, Garcea RL. Seroepidemiology of human polyomaviruses. PLoS Pathog. 2009;5:e1000363. https://doi.org/10.1371/journal.ppat.1000363.CrossRefPubMedPubMedCentral

15.

Zhang C, Liu F, He Z, Deng Q, Pan Y, Liu Y, Zhang C, Ning T, Guo C, Liang Y, Xu R, Zhang L, Cai H, Ke Y. Seroprevalence of Merkel cell polyomavirus in the general rural population of Anyang. China PLoS One. 2014;9:e106430. https://doi.org/10.1371/journal.pone.0106430.CrossRefPubMed

16.

Martel-Jantin C, Pedergnana V, Nicol JT, Leblond V, Tregouet DA, Tortevoye P, Plancoulaine S, Coursaget P, Touze A, Abel L. Gessain, A. Merkel cell polyomavirus infection occurs during early childhood and is transmitted between siblings. J Clin Virol 2013;58:288–291. doi: https://doi.org/10.1016/j.jcv.2013.06.004.

17.

Nicol JT, Robinot R, Carpentier A, Carandina G, Mazzoni E, Tognon M, Touze A, Coursaget P. Age-specific seroprevalences of merkel cell polyomavirus, human polyomaviruses 6, 7, and 9, and trichodysplasia spinulosa-associated polyomavirus. Clin Vaccine Immunol. 2013;20:363–8. https://doi.org/10.1128/CVI.00438-12.CrossRefPubMedPubMedCentral

18.

Kamminga S. E van der Meijden E, Feltkamp MCW. Zaaijer HL Seroprevalence of fourteen human polyomaviruses determined in blood donors PLoS One. 2018;13:e0206273. https://doi.org/10.1371/journal.pone.0206273.CrossRefPubMed

19.

Liu W, Yang R, Payne AS, Schowalter RM, Spurgeon ME, Lambert PF, Xu X, Buck CB, You J. Identifying the target cells and mechanisms of Merkel cell Polyomavirus infection. Cell Host Microbe. 2016;19:775–87. https://doi.org/10.1016/j.chom.2016.04.024.CrossRefPubMedPubMedCentral

20.

RM RS, Pastrana DV, Pumphrey KA, Moyer AL, Buck CB. Merkel cell polyomavirus and two previously unknown polyomaviruses are chronically shed from human skin. Cell Host Microbe. 2010;7:509–15. https://doi.org/10.1016/j.chom.2010.05.006.CrossRef

21.

Foulongne V, Sauvage V, Hebert C, Dereure O, Cheval J, Gouilh MA, Pariente K, Segondy M, Burguiere A, Manuguerra JC, Caro V, Eloit M. Human skin microbiota: high diversity of DNA viruses identified on the human skin by high throughput sequencing. PLoS One. 2012;7:e38499. https://doi.org/10.1371/journal.pone.0038499.CrossRefPubMedPubMedCentral

22.

Hashida Y, Kamioka M, Tanaka M, Hosokawa S, Murakami M, Nakajima K, Kikuchi H, Fujieda M, Sano S, Daibata M. Ecology of Merkel cell Polyomavirus in healthy skin among individuals in an Asian cohort. J Infect Dis. 2016;213:1708–16. https://doi.org/10.1093/infdis/jiw040.CrossRefPubMed

23.

Csoboz B, Rasheed K, Sveinbjørnsson U. Moens U. Merkel cell polyomavirus and non-Merkel cell carcinomas: guilty or circumstantial evidence? APMIS. 2020; [Online ahead of print]. https://doi.org/10.1111/apm.13019.

24.

Moens U, Van Ghelue M, Ludvigsen M, Korup-Schulz S, Ehlers B. The early and late promoters of BKPyV, MCPyV, TSPyV, and HPyV12 are among the strongest of all known human polyomaviruses in 10 different cell lines. J Gen Virol. 2015;96:2293–303. https://doi.org/10.1099/vir.0.000181.CrossRefPubMed

25.

Moens U, Song X, Van Ghelue M, Lednicky JA, Ehlers, B. A Role of Sp1 Binding Motifs in Basal and Large T-Antigen-Induced Promoter Activities of Human Polyomavirus HPyV9 and Its Variant UF-1. Int J Mol Sci. 2017;18:pii: E2414. doi: https://doi.org/10.3390/ijms18112414.

26.

Song X, Van Ghelue M, Ludvigsen M, Nordbo SA, Ehlers B, Moens U. Characterization of the non-coding control region of polyomavirus KI isolated from nasopharyngeal samples from patients with respiratory symptoms or infection and from blood from healthy blood donors in Norway. J Gen Virol. 2016;97:1647–57. https://doi.org/10.1099/jgv.0.000473.CrossRefPubMed

27.

Gosert R, Rinaldo CH, Funk GA, Egli A, Ramos E, Drachenberg CB, Hirsch HH. Polyomavirus BK with rearranged noncoding control region emerge in vivo in renal transplant patients and increase viral replication and cytopathology. J Exp Med. 2008;205:841–52. https://doi.org/10.1084/jem.20072097.CrossRefPubMedPubMedCentral

28.

Gosert R, Kardas P, Major EO, Hirsch HH. Rearranged JC virus noncoding control regions found in progressive multifocal leukoencephalopathy patient samples increase virus early gene expression and replication rate. J Virol. 2010;84:10448–56. https://doi.org/10.1128/JVI.00614-10.CrossRefPubMedPubMedCentral

29.

Ajuh ET, Wu Z, Kraus E, Weissbach FH, Bethge T, Gosert R, Fisher N, Hirsch HH. Novel human polyomavirus noncoding control region differ in bidirectional gene expression according to host cell, large T-antigen expression, and clinically occurring rearrangements. J Virol. 2018;92:e02231–17. https://doi.org/10.1128/JVI.02231-17.CrossRefPubMedPubMedCentral

30.

Barcena-Panero A, Echevarria JE, Van Ghelue M, Fedele G, Royuela E, Gerits N, Moens U. BK polyomavirus with archetypal and rearranged non-coding control regions is present in cerebrospinal fluids from patients with neurological complications. J Gen Virol. 2012;93:1780–94. https://doi.org/10.1099/vir.0.042143-0.CrossRefPubMed

31.

Van Loy T, Thys K, Ryschkewitsch C, Lagatie O, Monaco MC, Major EO, Tritsmans L, Stuyver LJ. JC virus quasispecies analysis reveals a complex viral population underlying progressive multifocal leukoencephalopathy and supports viral dissemination via the hematogenous route. J Virol. 2015;89:1340–7. https://doi.org/10.1128/JVI.02565-14.CrossRefPubMed

32.

L'Honneur AS, Leh H, Laurent-Tchenio F, Hazan U, Rozenberg F, Bury-Mone S. Exploring the role of NCCR variation on JC polyomavirus expression from dual reporter minicircles. PLoS One. 2018;13:e0199171. https://doi.org/10.1371/journal.pone.0199171.CrossRefPubMedPubMedCentral

33.

Takahashi K, Sekizuka T, Fukumoto H, Nakamichi K, Suzuki T, Sato Y, Hasegawa H, Kuroda M, Katano H. Deep-Sequence Identification and Role in Virus Replication of a JC Virus Quasispecies in Patients with Progressive Multifocal Leukoencephalopathy. J Virol. 2016;91:pii: e01335–e01316. doi: https://doi.org/10.1128/JVI.01335-16.

34.

Hashida Y, Higuchi T, Matsui K, Shibata Y, Nakajima K, Sano S, Daibata M. Genetic variability of the noncoding control region of cutaneous Merkel cell Polyomavirus: identification of geographically related genotypes. J Infect Dis. 2018;217:1601–11. https://doi.org/10.1093/infdis/jiy070.CrossRefPubMed

35.

Rasheed K, Abdulsalam I, Fismen S, Grimstad Ø, Sveinbjornsson B, Moens U. CCL17/TARC and CCR4 expression in Merkel cell carcinoma. Oncotarget. 2018;9:31432–47. https://doi.org/10.18632/oncotarget.25836.

36.

Pannuti A, Pascucci A, La Mantia G, Fisher-Fantuzzi L, Vesco C, Lania L. Trans-activation of cellular and viral promoters by a transforming nonkaryophilic simian virus 40 large T antigen. J Virol. 1987;61:1296–9.CrossRef

37.

Harrison C, Jiang T, Banerjee P, Meinke G, D'Abramo CM, Schaffhausen B, Bohm A. Polyomavirus large T antigen binds symmetrical repeats at the viral origin in an asymmetrical manner. J Virol. 2013;87:13751–9. https://doi.org/10.1128/JVI.01740-13.CrossRefPubMedPubMedCentral

38.

Shuda M, Feng H, Kwun HJ, Rosen ST, Gjoerup O, Moore PS, Chang Y. T antigen mutations are a human tumor-specific signature for Merkel cell polyomavirus. Proc Natl Acad Sci U S A. 2008;105:16272–7. https://doi.org/10.1073/pnas.0806526105.CrossRefPubMedPubMedCentral

39.

Horvath KB, Pankovics P, Battyani Z, Kalman E, Reuter G. A probable etiological role of Merkel cell polyomavirus in the development of Merkel cell carcinoma. Orv Hetil. 2013;154:102–12. https://doi.org/10.1556/OH.2013.29525.CrossRefPubMed

40.

Rosen ST, Gould VE, Salwen HR, Herst CV, Le Beau MM, Lee I, Bauer K, Marder RJ, Andersen R, Kies MS, et al. Establishment and characterization of a neuroendocrine skin carcinoma cell line. Lab Investig. 1987;56:302–12.PubMed

41.

Guastafierro A, Feng H, Thant M, Kirkwood JM, Chang Y, Moore PS, Shuda M. Characterization of an early passage Merkel cell polyomavirus-positive Merkel cell carcinoma cell line, MS-1, and its growth in NOD scid gamma mice. J Virol Methods. 2013;187:6–14. https://doi.org/10.1016/j.jviromet.2012.10.001.CrossRefPubMed

42.

Kwun HJ, Chang Y, Moore PS. Protein-mediated viral latency is a novel mechanism for Merkel cell polyomavirus persistence. Proc Natl Acad Sci U S A. 2017;114:e4040–7. https://doi.org/10.1073/pnas.1703879114.CrossRefPubMedPubMedCentral

43.

Moens U, Krumbholz A, Ehlers B, Zell R, Johne R, Calvignac-Spencer S, Lauber C. Biology, evolution, and medical importance of polyomaviruses: an update. Infect Genet Evol. 2017;54:18–38. https://doi.org/10.1016/j.meegid.2017.06.011.CrossRefPubMed

44.

Hashida Y, Imajoh M, Nemoto Y, Kamioka M, Taniguchi A, Taguchi T, Kume M, Orihashi K, Daibata M. Detection of Merkel cell polyomavirus with a tumour-specific signature in non-small cell lung cancer. Br J Cancer. 2013;108:629–37. https://doi.org/10.1038/bjc.2012.567.CrossRefPubMedPubMedCentral

45.

Pantulu ND, Pallasch CP, Kurz AK, Kassem A, Frenzel L, Sodenkamp S, Kvasnicka HM, Wendtner CM, Zur Hausen A. Detection of a novel truncating Merkel cell polyomavirus large T antigen deletion in chronic lymphocytic leukemia cells. Blood. 2010;116:5280–4. https://doi.org/10.1182/blood-2010-02-269829.CrossRefPubMed

46.

Velasquez C, Amako Y, Harold A, Toptan T, Chang Y, Shuda M. Characterization of a Merkel cell Polyomavirus-positive Merkel cell carcinoma cell line CVG-1. Front Microbiol. 2018;9:713. https://doi.org/10.3389/fmicb.2018.00713.

Title: Promoter activity of Merkel cell Polyomavirus variants in human dermal fibroblasts and a Merkel cell carcinoma cell line
Authors: Ibrahim Abdulsalam
Kashif Rasheed
Baldur Sveinbjørnsson
Bernhard Ehlers
Ugo Moens
Publication date: 01-12-2020
Publisher: BioMed Central
Keyword: Merkel Cell Carcinoma
Published in: Virology Journal / Issue 1/2020
Electronic ISSN: 1743-422X
DOI: https://doi.org/10.1186/s12985-020-01317-x

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Promoter activity of Merkel cell Polyomavirus variants in human dermal fibroblasts and a Merkel cell carcinoma cell line

Abstract

Background

Methods

Results

Conclusion

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Abstract

Background

Methods

Results

Conclusion

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