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Glycan-Based Near-infrared Fluorescent (NIRF) Imaging of Gastrointestinal Tumors: a Preclinical Proof-of-Concept In Vivo Study.
Molecular Imaging and Biology ( IF 3.1 ) Pub Date : 2020-08-11 , DOI: 10.1007/s11307-020-01522-8
Ruben D Houvast 1 , Victor M Baart 1 , Shadhvi S Bhairosingh 1 , Robert A Cordfunke 2 , Jia Xin Chua 3 , Mireille Vankemmelbeke 3 , Tina Parsons 3 , Peter J K Kuppen 1 , Lindy G Durrant 3, 4 , Alexander L Vahrmeijer 1 , Cornelis F M Sier 1, 5
Affiliation  

Purpose

Aberrantly expressed glycans in cancer are of particular interest for tumor targeting. This proof-of-concept in vivo study aims to validate the use of aberrant Lewis glycans as target for antibody-based, real-time imaging of gastrointestinal cancers.

Procedures

Immunohistochemical (IHC) staining with monoclonal antibody FG88.2, targeting Lewisa/c/x, was performed on gastrointestinal tumors and their healthy counterparts. Then, FG88.2 and its chimeric human/mouse variant CH88.2 were conjugated with near-infrared fluorescent (NIRF) IRDye 800CW for real-time imaging. Specific binding was evaluated in vitro on human gastrointestinal cancer cell lines with cell-based plate assays, flow cytometry, and immune-fluorescence microscopy. Subsequently, mice bearing human colon and pancreatic subcutaneous tumors were imaged in vivo after intravenous administration of 1 nmol (150 μg) CH88.2-800CW with the clinical Artemis NIRF imaging system using the Pearl Trilogy small animal imager as reference. One week post-injection of the tracer, tumors and organs were resected and tracer uptake was analyzed ex vivo.

Results

IHC analysis showed strong FG88.2 staining on colonic, gastric, and pancreatic tumors, while staining on their normal tissue counterparts was limited. Next, human cancer cell lines HT-29 (colon) and BxPC-3 and PANC-1 (both pancreatic) were identified as respectively high, moderate, and low Lewisa/c/x-expressing. Using the clinical NIRF camera system for tumor-bearing mice, a mean tumor-to-background ratio (TBR) of 2.2 ± 0.3 (Pearl: 3.1 ± 0.8) was observed in the HT-29 tumors and a TBR of 1.8 ± 0.3 (Pearl: 1.9 ± 0.5) was achieved in the moderate expression BxPC-3 model. In both models, tumors could be adequately localized and delineated by NIRF for up to 1 week. Ex vivo analysis confirmed full tumor penetration of the tracer and low fluorescence signals in other organs.

Conclusions

Using a novel chimeric Lewisa/c/x-targeting tracer in combination with a clinical NIRF imager, we demonstrate the potential of targeting Lewis glycans for fluorescence-guided surgery of gastrointestinal tumors.



中文翻译:

胃肠道肿瘤的基于聚糖的近红外荧光 (NIRF) 成像:一项临床前概念验证体内研究。

目的

癌症中异常表达的聚糖对肿瘤靶向特别感兴趣。这项概念验证体内研究旨在验证异常路易斯聚糖作为胃肠道癌症基于抗体的实时成像目标的使用。

程序

使用靶向 Lewis a/c/x 的单克隆抗体 FG88.2对胃肠道肿瘤及其健康对应物进行免疫组织化学 (IHC) 染色。然后,FG88.2 及其嵌合人/小鼠变体 CH88.2 与近红外荧光 (NIRF) IRDye 800CW 结合进行实时成像。使用基于细胞的平板检测、流式细胞术和免疫荧光显微镜对人胃肠癌细胞系进行体外特异性结合评估。随后,携带人结肠和胰腺皮下肿瘤的小鼠在体内成像静脉注射 1 nmol (150 μg) CH88.2-800CW 后,临床 Artemis NIRF 成像系统使用 Pearl Trilogy 小动物成像仪作为参考。注射示踪剂一周后,切除肿瘤和器官,并在体外分析示踪剂摄取。

结果

IHC 分析显示 FG88.2 对结肠、胃和胰腺肿瘤有很强的染色,而对正常组织对应物的染色有限。接下来,人类癌细胞系 HT-29(结肠)和 BxPC-3 和 PANC-1(均为胰腺)分别被鉴定为高、中和低表达 Lewis a/c/x。使用用于荷瘤小鼠的临床 NIRF 相机系统,在 HT-29 肿瘤中观察到平均肿瘤与背景比 (TBR) 为 2.2 ± 0.3 (Pearl: 3.1 ± 0.8),TBR 为 1.8 ± 0.3 ( Pearl:1.9 ± 0.5) 在中等表达 BxPC-3 模型中实现。在这两种模型中,NIRF 可以充分定位和描绘肿瘤长达 1 周。离体分析证实了示踪剂的完全肿瘤渗透和其他器官中的低荧光信号。

结论

使用新型嵌合 Lewis a/c/x靶向示踪剂与临床 NIRF 成像仪相结合,我们证明了靶向 Lewis 聚糖用于胃肠道肿瘤的荧光引导手术的潜力。

更新日期:2020-08-11
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