Improved ex vivo fluorescence imaging of human head and neck cancer using the peptide tracer TPP-IRDye800 targeting membrane-bound Hsp70 on tumor cells

使用肽示踪剂 TPP-IRDye800 靶向肿瘤细胞上的膜结合 Hsp70,改善人类头颈癌的体外荧光成像

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作者:Katharina L K Holzmann, Johanna L Wolf, Stefan Stangl, Philipp Lennartz, Atsuko Kasajima, Carolin Mogler, Bernhard Haller, Eva-Vanessa Ebert, Daniel Jira, Maren L A Lauterbach, Franziska von Meyer, Leonhard Stark, Leonie Mauch, Benedikt Schmidl, Barbara Wollenberg, Gabriele Multhoff #, Markus Wirth 

Background

The primary goal of surgery in HNSCC is the complete resection of tumor cells with maximum preservation of normal tissue. The membrane Hsp70-targeting fluorescence labelled peptide TPP-IRDye800 represents a promising tool for real-time intraoperative tumor visualization, enabling the detection of true tumor margins, critical isles of high-grade dysplasia and LN metastases.

Conclusion

TPP-IRDye800 is a promising fluorescence imaging probe for HNSCC.

Methods

Membrane Hsp70 (mHsp70) expression on HNSCC cell lines and primary HNSCC was determined by flow cytometry and fluorescence microscopy using FITC-conjugated mAb cmHsp70.1 and TPP. TPP-IRDye800 was sprayed on freshly resected tumor material of immunohistochemically confirmed HNSCC and LN metastases for tumor imaging. TBRs were compared using TPP-IRDye800 and Cetuximab-IRDye680, recognizing EGFR.

Results

mHsp70 expressing HNSCC cells specifically bind and internalize TPP in vitro. The TBR (2.56 ± 0.39) and AUC [0.98 CI, 0.95-1.00 vs. 0.91 CI, 0.85-0.97] of TPP-IRDye800 on primary HNSCC was significantly higher than Cetuximab-IRDye680 (1.61 ± 0.39) (p = 0.0068) and TPP-IRDye800 provided a superior tumor delineation. Fluorescence imaging showed higher AUC values than a visual inspection by surgeons [0.97 CI, 0.94-1.00 vs. 0.92 CI, 0.88-0.97] (p = 0.048). LN metastases could be visualized using TPP-IRDye800. Real-time tissue delineation was confirmed using the clinically applied KARL-STORZ imaging system.

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