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Published in: Annals of Surgical Oncology 12/2016

Open Access 01-11-2016 | Head and Neck Oncology

Intraoperative Sentinel Lymph Node Evaluation: Implications of Cytokeratin 19 Expression for the Adoption of OSNA in Oral Squamous Cell Carcinoma

Authors: Richard Shaw, MD, FRCS, FDS, Anders Christensen, MD, Kapil Java, MRCS, Rehab El Maddani, BDS, MRes, Triantafillos Liloglou, BSc, PhD, Triantafyllou Asterios, PhD, FRCPath, Christian von Buchwald, MD, DMSc, Irene Wessel, MD, PhD, Katalin Kiss, MD, Andreas Kjaer, MD, PhD, DMSc, Giedrius Lelkaitis, MD, Anna Long, BSc, MSc, Janet Risk, BSc, PhD, Max Robinson, PhD

Published in: Annals of Surgical Oncology | Issue 12/2016

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Abstract

Background

Intraoperative analysis of sentinel lymph nodes would enhance the care of early-stage oral squamous cell carcinoma (OSCC). We determined the frequency and extent of cytokeratin 19 (CK19) expression in OSCC primary tumours and surrounding tissues to explore the feasibility of a “clinic-ready” intraoperative diagnostic test (one step nucleic acid amplification—OSNA, sysmex).

Methods

Two cohorts were assembled: cohort 1, OSCC with stage and site that closely match cases suitable for sentinel lymph node biopsy (SLNB); cohort 2, HNSCC with sufficient fresh tumour tissue available for the OSNA assay (>50 mg). CK19 assays included qRT-PCR, RNA in situ hybridisation (ISH), and immunohistochemistry (IHC), as well as OSNA.

Results

CK19 mRNA expression was detected with variable sensitivity, depending on method, in 60–80% of primary OSCC tumours, while protein expression was observed in only 50% of tumours. Discordance between different techniques indicated that OSNA was more sensitive than qRT-PCR or RNA-ISH, which in turn were more sensitive than IHC. OSNA results showed CK19 expression in 80% of primary cases, so if used for diagnosis of lymph node metastasis would lead to a false-negative result in 20% of patients with cervical lymph node metastases.

Conclusions

OSNA in its current form is not suitable for use in OSCC SLNB due to inadequate expression of the CK19 target in all case. However, the same assay technology would likely be very promising if applied using a more ubiquitous squamous epithelial target.
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Literature
1.
go back to reference 1. Pedersen NJ, Jensen DH, Hedback N, et al. Staging of early lymph node metastases with the sentinel lymph node technique and predictive factors in T1/T2 oral cavity cancer: a retrospective single-center study. Head Neck. 2016;38:1033–40.CrossRef 1. Pedersen NJ, Jensen DH, Hedback N, et al. Staging of early lymph node metastases with the sentinel lymph node technique and predictive factors in T1/T2 oral cavity cancer: a retrospective single-center study. Head Neck. 2016;38:1033–40.CrossRef
2.
go back to reference Schilling C, Stoeckli SJ, Haerle SK, et al. Sentinel European Node Trial (SENT): 3-year results of sentinel node biopsy in oral cancer. Eur J Cancer. 2015;51(18):2777–84.CrossRefPubMed Schilling C, Stoeckli SJ, Haerle SK, et al. Sentinel European Node Trial (SENT): 3-year results of sentinel node biopsy in oral cancer. Eur J Cancer. 2015;51(18):2777–84.CrossRefPubMed
3.
go back to reference D’Cruz AK, Vaish R, Kapre N, et al. Elective versus therapeutic neck dissection in node-negative oral cancer. N Engl J Med. 2015;373(6):521–9.CrossRefPubMed D’Cruz AK, Vaish R, Kapre N, et al. Elective versus therapeutic neck dissection in node-negative oral cancer. N Engl J Med. 2015;373(6):521–9.CrossRefPubMed
4.
go back to reference Civantos FJ, Moffat FL, Goodwin WJ. Lymphatic mapping and sentinel lymphadenectomy for 106 head and neck lesions: contrasts between oral cavity and cutaneous malignancy. Laryngoscope. 2006;112(3 Pt 2 Suppl 109):1–15. Civantos FJ, Moffat FL, Goodwin WJ. Lymphatic mapping and sentinel lymphadenectomy for 106 head and neck lesions: contrasts between oral cavity and cutaneous malignancy. Laryngoscope. 2006;112(3 Pt 2 Suppl 109):1–15.
5.
go back to reference Tschopp L, Nuyens M, Stauffer E, Krause T, Zbaren P. The value of frozen section analysis of the sentinel lymph node in clinically N0 squamous cell carcinoma of the oral cavity and oropharynx. Otolaryngol Head Neck Surg. 2005;132(1):99–102.CrossRefPubMed Tschopp L, Nuyens M, Stauffer E, Krause T, Zbaren P. The value of frozen section analysis of the sentinel lymph node in clinically N0 squamous cell carcinoma of the oral cavity and oropharynx. Otolaryngol Head Neck Surg. 2005;132(1):99–102.CrossRefPubMed
6.
go back to reference Alkureishi LW, Burak Z, Alvarez JA, et al. Joint practice guidelines for radionuclide lymphoscintigraphy for sentinel node localization in oral/oropharyngeal squamous cell carcinoma. Ann Surg Oncol. 2009;16(11):3190–210.CrossRefPubMedPubMedCentral Alkureishi LW, Burak Z, Alvarez JA, et al. Joint practice guidelines for radionuclide lymphoscintigraphy for sentinel node localization in oral/oropharyngeal squamous cell carcinoma. Ann Surg Oncol. 2009;16(11):3190–210.CrossRefPubMedPubMedCentral
7.
go back to reference Ferris RL, Xi L, Seethala RR, et al. Intraoperative qRT-PCR for detection of lymph node metastasis in head and neck cancer. Clin Cancer Res. 2011;17(7):1858–66.CrossRefPubMed Ferris RL, Xi L, Seethala RR, et al. Intraoperative qRT-PCR for detection of lymph node metastasis in head and neck cancer. Clin Cancer Res. 2011;17(7):1858–66.CrossRefPubMed
8.
go back to reference Garrel R, Dromard M, Costes V, et al. The diagnostic accuracy of reverse transcription-PCR quantification of cytokeratin mRNA in the detection of sentinel lymph node invasion in oral and oropharyngeal squamous cell carcinoma: a comparison with immunohistochemistry. Clin Cancer Res. 2006;12(8):2498–505.CrossRefPubMed Garrel R, Dromard M, Costes V, et al. The diagnostic accuracy of reverse transcription-PCR quantification of cytokeratin mRNA in the detection of sentinel lymph node invasion in oral and oropharyngeal squamous cell carcinoma: a comparison with immunohistochemistry. Clin Cancer Res. 2006;12(8):2498–505.CrossRefPubMed
10.
go back to reference Snook KL, Layer GT, Jackson PA, et al. Multicentre evaluation of intraoperative molecular analysis of sentinel lymph nodes in breast carcinoma. Br J Surg. 2011;98(4):527–35.CrossRefPubMed Snook KL, Layer GT, Jackson PA, et al. Multicentre evaluation of intraoperative molecular analysis of sentinel lymph nodes in breast carcinoma. Br J Surg. 2011;98(4):527–35.CrossRefPubMed
11.
go back to reference NICE. Intraoperative tests (RD 100i OSNA system and Metasin test) for detecting sentinel lymph node metastases in breast cancer. London:NICE; 2013. NICE. Intraoperative tests (RD 100i OSNA system and Metasin test) for detecting sentinel lymph node metastases in breast cancer. London:NICE; 2013.
12.
go back to reference Klingler S, Marchal F, Rauch P, et al. Using one-step nucleic acid amplification (OSNA) for intraoperative detection of lymph node metastasis in breast cancer patients avoids second surgery and accelerates initiation of adjuvant therapy. Ann Oncol. 2013;24(9):2305–9.CrossRefPubMed Klingler S, Marchal F, Rauch P, et al. Using one-step nucleic acid amplification (OSNA) for intraoperative detection of lymph node metastasis in breast cancer patients avoids second surgery and accelerates initiation of adjuvant therapy. Ann Oncol. 2013;24(9):2305–9.CrossRefPubMed
13.
go back to reference Yamauchi K, Fujioka Y, Kogashiwa Y, Kohno N. Quantitative expression study of four cytokeratins and p63 in squamous cell carcinoma of the tongue: suitability for sentinel node navigation surgery using one-step nucleic acid amplification. J Clin Pathol. 2011;64(10):875–9.CrossRefPubMed Yamauchi K, Fujioka Y, Kogashiwa Y, Kohno N. Quantitative expression study of four cytokeratins and p63 in squamous cell carcinoma of the tongue: suitability for sentinel node navigation surgery using one-step nucleic acid amplification. J Clin Pathol. 2011;64(10):875–9.CrossRefPubMed
14.
go back to reference Zhong LP, Chen WT, Zhang CP, Zhang ZY. Increased CK19 expression correlated with pathologic differentiation grade and prognosis in oral squamous cell carcinoma patients. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2007;104(3):377–84.CrossRefPubMed Zhong LP, Chen WT, Zhang CP, Zhang ZY. Increased CK19 expression correlated with pathologic differentiation grade and prognosis in oral squamous cell carcinoma patients. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2007;104(3):377–84.CrossRefPubMed
15.
go back to reference Shores CG, Yin X, Funkhouser W, Yarbrough W. Clinical evaluation of a new molecular method for detection of micrometastases in head and neck squamous cell carcinoma. Arch Otolaryngol Head Neck Surg. 2004;130(8):937–42.CrossRefPubMed Shores CG, Yin X, Funkhouser W, Yarbrough W. Clinical evaluation of a new molecular method for detection of micrometastases in head and neck squamous cell carcinoma. Arch Otolaryngol Head Neck Surg. 2004;130(8):937–42.CrossRefPubMed
16.
go back to reference Croner RS, Schellerer V, Demund H, et al. One step nucleic acid amplification (OSNA) - a new method for lymph node staging in colorectal carcinomas. J Trans Med. 2010;8:83.CrossRef Croner RS, Schellerer V, Demund H, et al. One step nucleic acid amplification (OSNA) - a new method for lymph node staging in colorectal carcinomas. J Trans Med. 2010;8:83.CrossRef
17.
go back to reference Yamamoto H, Sekimoto M, Oya M, et al. OSNA-based novel molecular testing for lymph node metastases in colorectal cancer patients: results from a multicenter clinical performance study in Japan. Ann Surg Oncol. 2011;18(7):1891–8.CrossRefPubMed Yamamoto H, Sekimoto M, Oya M, et al. OSNA-based novel molecular testing for lymph node metastases in colorectal cancer patients: results from a multicenter clinical performance study in Japan. Ann Surg Oncol. 2011;18(7):1891–8.CrossRefPubMed
18.
go back to reference Kumagai K, Yamamoto N, Miyashiro I, et al. Multicenter study evaluating the clinical performance of the OSNA assay for the molecular detection of lymph node metastases in gastric cancer patients. Gastric Cancer. 2014;17(2):273–80.CrossRefPubMed Kumagai K, Yamamoto N, Miyashiro I, et al. Multicenter study evaluating the clinical performance of the OSNA assay for the molecular detection of lymph node metastases in gastric cancer patients. Gastric Cancer. 2014;17(2):273–80.CrossRefPubMed
19.
go back to reference Yaguchi Y, Sugasawa H, Tsujimoto H, et al. One-step nucleic acid amplification (OSNA) for the application of sentinel node concept in gastric cancer. Ann Surg Oncol. 2011;18(8):2289–96.CrossRefPubMed Yaguchi Y, Sugasawa H, Tsujimoto H, et al. One-step nucleic acid amplification (OSNA) for the application of sentinel node concept in gastric cancer. Ann Surg Oncol. 2011;18(8):2289–96.CrossRefPubMed
20.
go back to reference Goda H, Nakashiro K, Oka R, et al. One-step nucleic acid amplification for detecting lymph node metastasis of head and neck squamous cell carcinoma. Oral Oncol. 2012;48(10):958–63.CrossRefPubMed Goda H, Nakashiro K, Oka R, et al. One-step nucleic acid amplification for detecting lymph node metastasis of head and neck squamous cell carcinoma. Oral Oncol. 2012;48(10):958–63.CrossRefPubMed
21.
go back to reference Matsuzuka T, Takahashi K, Kawakita D, et al. Intraoperative molecular assessment for lymph node metastasis in head and neck squamous cell carcinoma using one-step nucleic acid amplification (OSNA) assay. Ann Surg Oncol. 2012;19(12):3865–70.CrossRefPubMed Matsuzuka T, Takahashi K, Kawakita D, et al. Intraoperative molecular assessment for lymph node metastasis in head and neck squamous cell carcinoma using one-step nucleic acid amplification (OSNA) assay. Ann Surg Oncol. 2012;19(12):3865–70.CrossRefPubMed
22.
go back to reference Suzuki M, Matsuzuka T, Hashimoto Y, Ikeda M, Saijo S, Omori K. Diagnostic potential of 1-step nucleic acid amplification assay in patients with head and neck squamous cell carcinoma based on CK19 expression in a primary lesion. Head Neck. 2014;38:E239–45. Suzuki M, Matsuzuka T, Hashimoto Y, Ikeda M, Saijo S, Omori K. Diagnostic potential of 1-step nucleic acid amplification assay in patients with head and neck squamous cell carcinoma based on CK19 expression in a primary lesion. Head Neck. 2014;38:E239–45.
23.
go back to reference Shinohara M, Harada T, Nakamura S, Oka M, Tashiro H. Heterotopic salivary gland tissue in lymph nodes of the cervical region. Int J Oral Maxillofac Surg. 1992;21(3):166–71.CrossRefPubMed Shinohara M, Harada T, Nakamura S, Oka M, Tashiro H. Heterotopic salivary gland tissue in lymph nodes of the cervical region. Int J Oral Maxillofac Surg. 1992;21(3):166–71.CrossRefPubMed
24.
go back to reference Leon X, Sancho FJ, Garcia J, Sanudo JR, Orus C, Quer M. Incidence and significance of clinically unsuspected thyroid tissue in lymph nodes found during neck dissection in head and neck carcinoma patients. Laryngoscope. 2005;115(3):470–4.CrossRefPubMed Leon X, Sancho FJ, Garcia J, Sanudo JR, Orus C, Quer M. Incidence and significance of clinically unsuspected thyroid tissue in lymph nodes found during neck dissection in head and neck carcinoma patients. Laryngoscope. 2005;115(3):470–4.CrossRefPubMed
25.
go back to reference Takahashi K, Nakajima K, Shino M, Toyoda M, Takayasu Y, Chikamatsu K. [Prediction of postoperative lymph node metastasis with a molecular biological test in head and neck cancer]. Nihon Jibiinkoka Gakkai kaiho. 2015;118(2):135–9.CrossRefPubMed Takahashi K, Nakajima K, Shino M, Toyoda M, Takayasu Y, Chikamatsu K. [Prediction of postoperative lymph node metastasis with a molecular biological test in head and neck cancer]. Nihon Jibiinkoka Gakkai kaiho. 2015;118(2):135–9.CrossRefPubMed
Metadata
Title
Intraoperative Sentinel Lymph Node Evaluation: Implications of Cytokeratin 19 Expression for the Adoption of OSNA in Oral Squamous Cell Carcinoma
Authors
Richard Shaw, MD, FRCS, FDS
Anders Christensen, MD
Kapil Java, MRCS
Rehab El Maddani, BDS, MRes
Triantafillos Liloglou, BSc, PhD
Triantafyllou Asterios, PhD, FRCPath
Christian von Buchwald, MD, DMSc
Irene Wessel, MD, PhD
Katalin Kiss, MD
Andreas Kjaer, MD, PhD, DMSc
Giedrius Lelkaitis, MD
Anna Long, BSc, MSc
Janet Risk, BSc, PhD
Max Robinson, PhD
Publication date
01-11-2016
Publisher
Springer International Publishing
Published in
Annals of Surgical Oncology / Issue 12/2016
Print ISSN: 1068-9265
Electronic ISSN: 1534-4681
DOI
https://doi.org/10.1245/s10434-016-5337-6

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