Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Nuclear expression of AFF2 C-terminus is a sensitive and specific ancillary marker for DEK::AFF2 carcinoma of the sinonasal tract

Abstract

DEK::AFF2 carcinoma of the sinonasal tract is an emerging entity. The tumor is typically characterized by papillary proliferation of non-keratinizing squamous epithelial cells with monotonous cytologic features, which may mimic other sinonasal tumors. The confirmation of this gene fusion has thus far relied solely on next-generation sequencing, fluorescence in situ hybridization (FISH), or reverse transcription polymerase chain reaction (RT-PCR). This current study aimed to validate an immunohistochemical assay for AFF2 C-terminus as an ancillary marker. We first analyzed publicly available RNA sequencing data of sinonasal tumors from the national center for biotechnology information (NCBI) sequence read archive and identified 3 DEK::AFF2 carcinomas out of 28 sinonasal tumors. The gene expression of AFF2 was significantly higher in the fusion-positive cases compared to the wild-type tumors (p < 0.001), while DEK was not. We then optimized an immunohistochemical assay with an anti-AFF2 C-terminus antibody for ancillary diagnosis. Seventeen DEK::AFF2 carcinomas, including 11 cases with predominantly low-grade morphology and one showing glandular differentiation, as well as 78 DEK FISH-negative sinonasal tumors were evaluated by AFF2 immunohistochemistry (IHC). Sixteen of the 17 DEK::AFF2 carcinomas showed nuclear AFF2 expression in ≥30% of tumor cells, including one decalcified case that failed FISH and RT-PCR confirmation. The one case that was negative for AFF2 IHC in the tumor cells also lacked expression in the internal positive control. It was thus considered a failure of the IHC rather than a truly negative case and was excluded from the statistical analysis. All DEK FISH-negative sinonasal tumors were negative for nuclear AFF2 expression. The nuclear expression of AFF2 IHC showed 100% sensitivity and specificity for DEK::AFF2 carcinoma. Accordingly, AFF2 IHC is a highly sensitive and specific ancillary marker that distinguishes DEK-AFF2 carcinoma from the other sinonasal tumors with overlapping morphological features and may be an especially useful alternative for decalcified specimens.

Your institute does not have access to this article

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Fig. 1: RNA sequencing data mining.
Fig. 2: Illustration of the AFF2 immunohistochemistry detection of the DEK::AFF2 fusion protein.
Fig. 3: Morphologic spectrum of DEK::AFF2 carcinoma and the AFF2 immunohistochemistry (IHC).
Fig. 4: Morphology and AFF2 immunohistochemistry (IHC) of the DEK fluorescence in situ hybridization-negative sinonasal tumors (H&E, 200X; inset: AFF2 IHC, 400X).

Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

References

  1. Yang, W., Lee, K.W., Srivastava, R.M., Kuo, F., Krishna, C., Chowell, D. et al. Immunogenic neoantigens derived from gene fusions stimulate T cell responses. Nat. Med. 25, 767–775 (2019).

  2. Todorovic, E., Truong, T., Eskander, A., Lin, V., Swanson, D., Dickson, B.C. et al. Middle ear and temporal bone nonkeratinizing squamous cell carcinomas with DEK-AFF2 fusion: an emerging entity. Am. J. Surg. Pathol. 44, 1244–1250 (2020).

  3. Bishop, J.A., Gagan, J., Paterson, C., McLellan, D., Sandison, A. Nonkeratinizing Squamous Cell Carcinoma of the Sinonasal Tract With DEK-AFF2: Further Solidifying an Emerging Entity. Am. J. Surg. Pathol. 45, 718–720 (2021).

  4. Kuo, Y.J., Lewis, J.S., Jr, Zhai C., Chen, Y.A., Chernock, R.D., Hsieh, M.S. et al. DEK-AFF2 fusion-associated papillary squamous cell carcinoma of the sinonasal tract: clinicopathologic characterization of seven cases with deceptively bland morphology. Mod. Pathol. 34, 1820–1830 (2021).

  5. Rooper LM, Agaimy A, Dickson BC, Dueber JC, Eberhart CG, Gagan J, et al. DEK-AFF2 Carcinoma of the Sinonasal Region and Skull Base: Detailed Clinicopathologic Characterization of a Distinctive Entity. Am. J. Surg. Pathol. 45, 1682–1693 (2021).

  6. Waldmann, T., Scholten, I., Kappes, F., Hu, H.G., Knippers, R. The DEK protein-an abundant and ubiquitous constituent of mammalian chromatin. Gene 343, 1–9 (2004).

  7. Riveiro-Falkenbach, E., Soengas, M.S. Control of tumorigenesis and chemoresistance by the DEK oncogene. Clin. Cancer Res. 16, 2932–2938 (2010).

  8. Teng, Y., Lang, L., Jauregui, C.E. The Complexity of DEK Signaling in Cancer Progression. Curr. Cancer Drug Targets 18, 256–265 (2018).

  9. Ishida, K., Nakashima, T., Shibata, T., Hara, A., Tomita, H. Role of the DEK oncogene in the development of squamous cell carcinoma. Int. J. Clin. Oncol. 25, 1563–1569 (2020).

  10. Wise-Draper, T.M., Draper, D.J., Gutkind, J.S., Molinolo, A.A., Wikenheiser-Brokamp, K.A., Wells, S.I. Future directions and treatment strategies for head and neck squamous cell carcinomas. Transl. Res. 160, 167–177 (2012).

  11. Adams, A.K., Hallenbeck, G.E., Casper, K.A., Patil, Y.J., Wilson, K.M., Kimple, R.J., et al. DEK promotes HPV-positive and -negative head and neck cancer cell proliferation. Oncogene 34, 868–877 (2015).

  12. Nakashima, T., Tomita, H., Hirata, A., Ishida, K., Hisamatsu, K., Hatano, Y. et al. Promotion of cell proliferation by the proto-oncogene DEK enhances oral squamous cell carcinogenesis through field cancerization. Cancer Med. 6, 2424–2439 (2017).

  13. Bensaid, M., Melko, M., Bechara, E.G., Davidovic, L., Berretta, A., Catania, M.V. et al. FRAXE-associated mental retardation protein (FMR2) is an RNA-binding protein with high affinity for G-quartet RNA forming structure. Nucleic Acids Res. 37, 1269–1279 (2009).

  14. Ge, H., Liu, K., Juan, T., Fang, F., Newman, M., Hoeck, W. FusionMap: detecting fusion genes from next-generation sequencing data at base-pair resolution. Bioinformatics 27, 1922–1928 (2011).

  15. Robinson, J.T., Thorvaldsdottir, H., Wenger, A.M., Zehir, A., Mesirov, J.P. Variant Review with the Integrative Genomics Viewer. Cancer Res. 77, e31–e34 (2017).

  16. Dobin, A., Davis, C.A., Schlesinger, F., Drenkow, J., Zaleski, C., Jha, S., et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics 29, 15–21 (2013).

  17. Anders, S., Pyl, P.T., Huber, W. HTSeq-a Python framework to work with high-throughput sequencing data. Bioinformatics 31, 166–169 (2015).

  18. Bell, D., Bell, A.H., Kupferman, M.E., Prieto, V.G., Weber, R.S., Hanna, E.Y. Comparative transcriptome analysis of sinonasal inverted papilloma and associated squamous cell carcinoma: Out-HOXing developmental genes. Head Neck 41, 3090–3104 (2019).

  19. Saab-Chalhoub, M.W., Guo, X., Shi, Q., Chernock, R.D., Lewis, J.S., Jr Low Grade Papillary Sinonasal (Schneiderian) Carcinoma: A Series of Five Cases of a Unique Malignant Neoplasm with Comparison to Inverted Papilloma and Conventional Nonkeratinizing Squamous Cell Carcinoma. Head Neck Pathol. 15, 1221–1234 (2021).

  20. Lewis, J.S., Jr., Chernock, R.D., Haynes, W., El-Mofty, S.K. Low-grade papillary schneiderian carcinoma, a unique and deceptively bland malignant neoplasm: report of a case. Am. J. Surg. Pathol. 39, 714–721 (2015).

  21. Zhai, C., Wang, H., Li, S., Wang, D. Clinicopathological analysis of low-grade papillary Schneiderian carcinoma: report of five new cases and review of the literature. Histopathology 79, 370–380 (2021).

  22. Bishop, J.A., Brandwein-Gensler, M., Nicolai, P., Steens, S., Syjanen, S., Westra, W.H. Non-keratinizing squamous cell carcinoma, (eds El-Naggar, A.K., Chan, J.K.C., Grandis, J.R., Takata, T., Slootweg, P.J.). WHO Classification of Head and Neck Tumours. 4th edn. 15–17 (IARC: Lyon, 2017).

  23. Bishop, J.A., Guo, T.W., Smith, D.F., Wang, H., Ogawa, T., Pai, S.I. et al. Human papillomavirus-related carcinomas of the sinonasal tract. Am. J. Surg. Pathol. 37, 185–192 (2013).

  24. El-Mofty, S.K., Lu, D.W. Prevalence of high-risk human papillomavirus DNA in nonkeratinizing (cylindrical cell) carcinoma of the sinonasal tract: a distinct clinicopathologic and molecular disease entity. Am. J. Surg. Pathol. 29, 1367–1372 (2005).

  25. Larque, A.B., Hakim, S., Ordi, J., Nadal, A., Diaz, A., del Pino, M., et al. High-risk human papillomavirus is transcriptionally active in a subset of sinonasal squamous cell carcinomas. Mod. Pathol. 27, 343–351 (2014).

  26. Nudell, J., Chiosea, S., Thompson, L.D. Carcinoma ex-Schneiderian papilloma (malignant transformation): a clinicopathologic and immunophenotypic study of 20 cases combined with a comprehensive review of the literature. Head Neck Pathol. 8, 269–286 (2014).

  27. Bubola, J., MacMillan, C.M., Weinreb, I., Witterick, I., Swanson, D., Zhang, L., et al. A Poorly Differentiated Non-keratinizing Sinonasal Squamous Cell Carcinoma with a Novel ETV6-TNFRSF8 Fusion Gene. Head Neck Pathol. 15, 1284–1288 (2021).

  28. Agaimy, A., Hartmann, A., Antonescu, C.R., Chiosea, S.I., El-Mofty, S.K., Geddert, H. et al. SMARCB1 (INI-1)-deficient Sinonasal Carcinoma: A Series of 39 Cases Expanding the Morphologic and Clinicopathologic Spectrum of a Recently Described Entity. Am. J. Surg. Pathol. 41, 458–471 (2017).

  29. French, C.A. NUT Carcinoma: Clinicopathologic features, pathogenesis, and treatment. Pathol. Int. 68, 583–595 (2018).

  30. Bishop, J.A., Alaggio, R., Zhang, L., Seethala, R.R., Antonescu, C.R. Adamantinoma-like Ewing family tumors of the head and neck: a pitfall in the differential diagnosis of basaloid and myoepithelial carcinomas. Am. J. Surg. Pathol. 39, 1267–1274 (2015).

Download references

Acknowledgements

The authors would like to thank the Biobank, Taipei Veterans General Hospital for assistance with sample preparation in this study. This study was presented in part at the annual meeting of the United States and Canadian Academy of Pathology on March 21st, 2022, Los Angeles, CA, USA.

Funding

The study was supported by the research grants from Taipei Veterans General Hospital (V110B-019) and the Ministry of Science and Technology, Taiwan (MOST110-2320-B-075-003-MY3).

Author information

Authors and Affiliations

Authors

Contributions

Y.J.K. and J.F.H. performed the study concept and design, analyzed the data, and wrote the paper; Y.C.Y. and J.F.H. conducted bioinformatics analysis of the RNAseq data; Y.J.K., J.S.L., T.T., R.D.C., C.Z., Y.A.C., T.H., C.K.L., Q.S., J.M.V., A.B.G., Y.H.C., M.S.H., A.H., and J.F.H. provided specimen acquisition and clinical data; J.F.H. supported immunohistochemistry and molecular methodology; J.S.L., R.D.C., and I.W. performed critical review and editing. All authors read and approved the final paper.

Corresponding author

Correspondence to Jen-Fan Hang.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

The study was approved by the institutional review board (IRB) of Taipei Veterans General Hospital (IRB no.: 2020-12-012CC).

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Kuo, YJ., Lewis, J.S., Truong, T. et al. Nuclear expression of AFF2 C-terminus is a sensitive and specific ancillary marker for DEK::AFF2 carcinoma of the sinonasal tract. Mod Pathol (2022). https://doi.org/10.1038/s41379-022-01117-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1038/s41379-022-01117-4

Search

Quick links