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Background <p>In surgical treatment of head and neck squamous cell carcinoma (HNSCC), resection with adequate tumor-free margins is a fundamental challenge. <i>Epithelial cell adhesion molecule</i> (EpCAM/CD326) is substantially overexpressed in HNSCC representing a potential target for fluorescence-guided delineation of HNSCC.</p> Materials and methods <p>EpCAM expression was assessed in vitro on squamous cell carcinomas of the upper aerodigestive tract (SCC-UADT) and fibroblasts employing immunostaining with anti-EpCAM antibodies VU1D9 and MT201/adecatumumab (clinically validated). Whole tumor analyses of EpCAM expression were conducted in patient samples with primary and recurrent HNSCC. Confocal microscopy was used for EpCAM expression analyses upon immunostaining in cultured patient HNSCC resection samples using IRDye800CW-labeled MT201.</p> Results <p>Immunostaining with VU1D9 and MT201 revealed a high, consistent, and specific expression of EpCAM on SCC-UADT in vitro. Human whole tumor analyses showed high and consistent EpCAM expression (primary: 9/9 &gt; = 80%; recurrent: 6/9 &gt; = 80%) and significantly elevated near-infrared fluorescence intensities for IRDye800CW-VU1D9 and IRDye800CW-MT201 in HNSCC as compared to non-malignant tissue (2–7 fold vs. mucosa; 6–20 fold vs. HNSCC-associated stroma). Translational postoperative immunostaining of live cultured patient HNSCC samples using the clinically validated IRDye800CW-MT201/adecatumumab enabled microscopical differentiation of HNSCC from adjacent non-malignant tissue (fluorescence intensity ratios HNSCC vs. mucosa: 7,66 ± 1,26; and vs. HNSCC-associated stroma: 50,97 ± 8,24)).</p> Conclusions <p>IRDye800CW-labeled anti-EpCAM antibodies allow experimental microscopic delineation of HNSCC from non-malignant tissue. As a future outlook, these probes might improve tumor resection in near-infrared fluorescence-guided surgery of primary and recurrent HNSCC.</p>

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Targeting EpCAM expression via near-infrared fluorescent antibodies enables microscopic delineation of primary and recurrent HNSCC

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年5月25日

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  • Bernd Uhl,
  • Dennis Eggert,
  • Bojan Smiljanov,
  • Katharina von Thun und Hohenstein,
  • Christoph Walz,
  • Gisela Kranz,
  • Sabina Schwenk-Zieger,
  • Florian Haring,
  • Joshua Luft,
  • Jiahang Song,
  • Vincent Holtmann,
  • Philipp Baumeister,
  • Christoph A. Reichel,
  • Olivier Gires,
  • Martin Canis,
  • Christian Betz

摘要

Background

In surgical treatment of head and neck squamous cell carcinoma (HNSCC), resection with adequate tumor-free margins is a fundamental challenge. Epithelial cell adhesion molecule (EpCAM/CD326) is substantially overexpressed in HNSCC representing a potential target for fluorescence-guided delineation of HNSCC.

Materials and methods

EpCAM expression was assessed in vitro on squamous cell carcinomas of the upper aerodigestive tract (SCC-UADT) and fibroblasts employing immunostaining with anti-EpCAM antibodies VU1D9 and MT201/adecatumumab (clinically validated). Whole tumor analyses of EpCAM expression were conducted in patient samples with primary and recurrent HNSCC. Confocal microscopy was used for EpCAM expression analyses upon immunostaining in cultured patient HNSCC resection samples using IRDye800CW-labeled MT201.

Results

Immunostaining with VU1D9 and MT201 revealed a high, consistent, and specific expression of EpCAM on SCC-UADT in vitro. Human whole tumor analyses showed high and consistent EpCAM expression (primary: 9/9 > = 80%; recurrent: 6/9 > = 80%) and significantly elevated near-infrared fluorescence intensities for IRDye800CW-VU1D9 and IRDye800CW-MT201 in HNSCC as compared to non-malignant tissue (2–7 fold vs. mucosa; 6–20 fold vs. HNSCC-associated stroma). Translational postoperative immunostaining of live cultured patient HNSCC samples using the clinically validated IRDye800CW-MT201/adecatumumab enabled microscopical differentiation of HNSCC from adjacent non-malignant tissue (fluorescence intensity ratios HNSCC vs. mucosa: 7,66 ± 1,26; and vs. HNSCC-associated stroma: 50,97 ± 8,24)).

Conclusions

IRDye800CW-labeled anti-EpCAM antibodies allow experimental microscopic delineation of HNSCC from non-malignant tissue. As a future outlook, these probes might improve tumor resection in near-infrared fluorescence-guided surgery of primary and recurrent HNSCC.