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

Sep 23, 2026

Uploaded by Adam

Surgical management of head and neck squamous cell carcinoma (HNSCC) depends on achieving tumor-free margins, yet defining boundaries remains challenging. EpCAM/CD326 is substantially overexpressed in HNSCC and is proposed as a target for fluorescence-guided delineation. Using immunostaining with clinically validated anti-EpCAM antibodies VU1D9 and MT201/adecatumumab and IRDye800CW labeling, EpCAM expression was evaluated in vitro and in whole tumor patient samples, including primary and recurrent disease. IRDye800CW probes enabled microscopic differentiation of HNSCC from adjacent non-malignant tissue and supported near-infrared surgical margin visualization.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages12

Reading Time30 min

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Abstract

Background

Materials and methods

Results

Conclusions

Introduction/Background

Uhl et al. BMC Cancer (2026) 26:671 BMC Cancer [https://doi.org/10.1186/s12885-026-16172-2](https://doi.org/10.1186/s12885-026-16172-2) RESEARCH Open Access Targeting EpCAM expression via near-infrared fluorescent antibodies enables microscopic delineation of primary and recurrent HNSCC Bernd Uhl 1,2,3,4,7*, Dennis Eggert5, Bojan Smiljanov2,3, Katharina von Thun und Hohenstein5, Christoph Walz6, Gisela Kranz2, Sabina Schwenk-Zieger2, Florian Haring2,3, Joshua Luft2,3, Jiahang Song2, Vincent Holtmann2, Philipp Baumeister2,7,8, Christoph A. Reichel2,3,7, Olivier Gires2, Martin Canis2,7,8† and Christian Betz5† Abstract 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 ofthe 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 EpCAMon 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 IRDye800CWVU1D9 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. Keywords HNSCC, EpCAM, Image-guided surgery, Fluorescence-guided surgery, Near-infrared, IRDye800 †Martin Canis and Christian Betz shared senior authorship. *Correspondence: Bernd Uhl [Bernd.Uhl@mri.tum.de](Bernd.Uhl@mri.tum.de) Full list of author information is available at the end of the article © The Author(s) 2026. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creativecommons.org/licenses/by/4.0/](http://creativecommons.org/licenses/by/4.0/.)[.](http://creativecommons.org/licenses/by/4.0/.) Uhl et al. BMC Cancer (2026) 26:671 Introduction/Background Head and neck cancer is the seventh most common cancer entity worldwide, which are mainly represente

FAQs

Why is delineating tumor-free margins important in HNSCC surgery?

Resection with adequate tumor-free margins is a fundamental challenge. It is associated with reduced cancer recurrence and lower patient morbidity and mortality.

What is the proposed biological target for fluorescence-guided delineation?

Epithelial cell adhesion molecule (EpCAM/CD326) is substantially overexpressed in HNSCC, making it a potential target for fluorescence-guided delineation.

How do the near-infrared fluorescent antibodies enable microscopic differentiation?

Clinically validated IRDye800CW-labeled MT201/adecatumumab allowed microscopical differentiation of HNSCC from adjacent non-malignant tissue by using fluorescence intensity ratios and image-based contrast.

Sep 23, 2026

Uploaded by Adam