使用靶向肿瘤细胞膜结合Hsp70的肽探针TPP-IRDye800进行人头颈癌的改良离体荧光成像

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

BRITISH JOURNAL OF CANCER

影响因子:6.8

分区:医学2区 Top / 肿瘤学2区

发表日期:2024 Dec

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

DOI: 10.1038/s41416-024-02872-8

摘要

HNSCC手术的主要目标是完全切除肿瘤细胞，同时最大限度地保留正常组织。膜Hsp70靶向的荧光标记肽TPP-IRDye800是一种具有潜力的实时术中肿瘤成像工具，能够检测真正的肿瘤边界、高级别异型增生的关键区域以及淋巴结转移。通过流式细胞术和荧光显微镜，使用FITC偶联的cmHsp70.1单克隆抗体（mAb）和TPP，对HNSCC细胞系及原发性HNSCC的膜Hsp70（mHsp70）表达进行了检测。将TPP-IRDye800喷涂在新鲜切除的免疫组化确认的HNSCC及淋巴结转移的肿瘤材料上进行肿瘤成像。通过TPP-IRDye800和识别EGFR的Cetuximab-IRDye680对比TBR（肿瘤对背景比值）。在体外，表达mHsp70的HNSCC细胞特异性结合并内吞TPP。TPP-IRDye800在原发性HNSCC上的TBR（2.56 ± 0.39）及AUC（[0.98 CI, 0.95-1.00]对[0.91 CI, 0.85-0.97]）明显高于Cetuximab-IRDye680（1.61 ± 0.39）（p = 0.0068），且提供了更优的肿瘤界定。荧光成像显示其AUC值高于外科医生的视觉检测[0.97 CI, 0.94-1.00对0.92 CI, 0.88-0.97]（p = 0.048）。淋巴结转移也能被TPP-IRDye800可视化。利用临床应用的KARL-STORZ成像系统验证了实时组织界定能力。TPP-IRDye800是一种有前景的HNSCC荧光成像探针。

Abstract

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