Verbeek Floris P R, van der Vorst Joost R, Tummers Quirijn R J G, Boonstra Martin C, de Rooij Karien E, Löwik Clemens W G M, Valentijn A Rob P M, van de Velde Cornelis J H, Choi Hak Soo, Frangioni John V, Vahrmeijer Alexander L
Department of Surgery, Leiden University Medical Center, Leiden, The Netherlands.
Ann Surg Oncol. 2014 Dec;21 Suppl 4(0 4):S528-37. doi: 10.1245/s10434-014-3524-x. Epub 2014 Feb 11.
Irradical tumor resections and iatrogenic ureteral injury remain a significant problem during lower abdominal surgery. The aim of the current study was to intraoperatively identify both colorectal tumors and ureters in subcutaneous and orthotopic animal models using cRGD-ZW800-1 and near-infrared (NIR) fluorescence.
The zwitterionic fluorophore ZW800-1 was conjugated to the tumor specific peptide cRGD (targeting integrins) and to the a-specific peptide cRAD. One nmol cRGD-ZW800-1, cRAD-ZW800-1, or ZW800-1 alone was injected in mice bearing subcutaneous HT-29 human colorectal tumors. Subsequently, cRGD-ZW800-1 was injected at dosages of 0.25 and 1 nmol in mice bearing orthotopic HT-29 tumors transfected with luciferase2. In vivo biodistribution and ureteral visualization were investigated in rats. Fluorescence was measured intraoperatively at several time points after probe administration using the FLARE imaging system.
Both subcutaneous and orthotopic tumors could be clearly identified using cRGD-ZW800-1. A significantly higher signal-to-background ratio was observed in mice injected with cRGD-ZW800-1 (2.42 ± 0.77) compared with mice injected with cRAD-ZW800-1 or ZW800-1 alone (1.21 ± 0.19 and 1.34 ± 0.19, respectively) when measured at 24 h after probe administration. The clearance of cRGD-ZW800-1 permitted visualization of the ureters and also generated minimal background fluorescence in the gastrointestinal tract.
This study appears to be the first to demonstrate both clear tumor demarcation and ureteral visualization after a single intravenous injection of a targeted NIR fluorophore. As a low dose of cRGD-ZW800-1 provided clear tumor identification, clinical translation of these results should be possible.
在腹部低位手术中,肿瘤切除不彻底和医源性输尿管损伤仍然是一个重大问题。本研究的目的是在皮下和原位动物模型中,使用cRGD-ZW800-1和近红外(NIR)荧光技术在术中识别结直肠肿瘤和输尿管。
两性离子荧光团ZW800-1与肿瘤特异性肽cRGD(靶向整合素)和α特异性肽cRAD偶联。将1 nmol的cRGD-ZW800-1、cRAD-ZW800-1或单独的ZW800-1注射到患有皮下HT-29人结直肠肿瘤的小鼠体内。随后,将0.25 nmol和1 nmol剂量的cRGD-ZW800-1注射到转染了荧光素酶2的原位HT-29肿瘤小鼠体内。在大鼠中研究体内生物分布和输尿管显影。使用FLARE成像系统在探针给药后的几个时间点进行术中荧光测量。
使用cRGD-ZW800-1可以清楚地识别皮下和原位肿瘤。在探针给药后24小时测量时,与单独注射cRAD-ZW800-1或ZW800-1的小鼠(分别为1.21±0.19和1.34±0.19)相比,注射cRGD-ZW800-1的小鼠观察到显著更高的信噪比(2.42±0.77)。cRGD-ZW800-1的清除允许输尿管显影,并且在胃肠道中产生的背景荧光也最小。
本研究似乎首次证明了单次静脉注射靶向近红外荧光团后,肿瘤边界清晰且输尿管显影。由于低剂量的cRGD-ZW800-1提供了清晰的肿瘤识别,这些结果应该有可能进行临床转化。