在CPC-APC小鼠模型中使用荧光团偶联的Claudin-1抗体增强结肠息肉的可视化。
Enhanced visualization of colonic polyps using a fluorophore-conjugated claudin-1 antibody in a CPC-APC mouse model.
作者信息
Liu Shanglei, Cox Kristin E, Amirfakhri Siamak, Jaiswal Sunidhi, Talib Sumbal, Primeaux Mark, Jih Lily J, Singh Satish K, Pisegna Joseph R, Mohs Aaron M, Hoffman Robert M, Batra Surinder K, Dhawan Punita, Bouvet Michael
机构信息
Department of Surgery, University of California San Diego, La Jolla, CA, United States.
Department of Surgery, University of California San Diego, La Jolla, CA, United States; Veteran's Affair San Diego Healthcare System, La Jolla, CA, United States.
出版信息
J Gastrointest Surg. 2025 Jun 13:102103. doi: 10.1016/j.gassur.2025.102103.
BACKGROUND
Early detection and removal of polyps for primary colon cancer prevention during colonoscopy are still fraught with significant miss rates. In the current study, we determined the ability of claudin-1 (CLDN1) antibodies conjugated to near-infrared fluorophores to target and visualize colonic polyps with high resolution in the CPC-APC mouse colonic polyp model.
METHODS
CPC-APC mice that developed distal colon polyps at 12 weeks of age were used in the present study. The formation of polyps was confirmed by mouse colonoscopy using the ColoView system (Karl Stortz). For polyp labeling, anti-CLDN1 and control immunoglobulin G (IgG) antibodies were conjugated to IRDye800 and 50, 100, and 150 μg doses were administered intravenouly to CPC-APC mice. Mice were sacrificed at different time points, and colons were harvested for imaging using the LI-COR Pearl Small Animal Imaging System. Polyp-to-background fluorescence intensity ratios (PBRs) were calculated for each polyp. Histology and immunohistochemistry (IHC) were performed on the polyps.
RESULTS
A total of 9 mice were used in the experiment, with a total of 45 polyps analyzed. The presence of observed polyps on gross examination was confirmed via histology. CLDN1-IRDye800 had a PBR of 7.6 ± 3.9, which was significantly higher than the nonspecific IgG-IRDye800 control (P =.0428). CLDN1-IRDye800 was further analyzed and overall had a similar range of PBR with 50, 100, and 150 μg injections at 72 h and had a statistically significant higher PBR of 8.9 ± 4.2 than the IgG-IRDye800 control (P =.0184) at 150 μg. Adenomatous polyps as small as 1 mm could be identified using CLDN1-IRDye800. Hematoxylin and eosin staining confirmed adenomatous polyps with high-grade dysplasia, and IHC confirmed the expression of CLDN1.
CONCLUSION
CLDN1-IRDye800 enabled polyp visualization with ultrahigh-resolution fluorescence imaging technology in CPC-APC mice. Polyps as small as 1 mm could be visualized, indicating clinical potential to overcome the current limitations of colonic polyp detection.
背景
在结肠镜检查期间，为预防原发性结肠癌而进行的息肉早期检测和切除，其漏诊率仍然很高。在本研究中，我们在CPC-APC小鼠结肠息肉模型中，确定了与近红外荧光团偶联的紧密连接蛋白-1（CLDN1）抗体靶向并高分辨率可视化结肠息肉的能力。
方法
本研究使用了12周龄时出现远端结肠息肉的CPC-APC小鼠。使用ColoView系统（卡尔·史托斯）通过小鼠结肠镜检查确认息肉的形成。为了标记息肉，将抗CLDN1抗体和对照免疫球蛋白G（IgG）抗体与IRDye800偶联，并以50、100和150μg的剂量静脉注射给CPC-APC小鼠。在不同时间点处死小鼠，采集结肠并使用LI-COR Pearl小动物成像系统进行成像。计算每个息肉的息肉与背景荧光强度比（PBR）。对息肉进行组织学和免疫组织化学（IHC）检查。
结果
实验共使用了9只小鼠，共分析了45个息肉。通过组织学证实了大体检查中观察到的息肉的存在。CLDN1-IRDye800的PBR为7.6±3.9，显著高于非特异性IgG-IRDye800对照（P = 0.0428）。对CLDN1-IRDye800进行了进一步分析，总体而言，在72小时时，50、100和150μg注射剂量的PBR范围相似，在150μg时，其PBR为8.9±4.2，显著高于IgG-IRDye800对照（P = 0.0184）。使用CLDN1-IRDye800可以识别小至1mm的腺瘤性息肉。苏木精和伊红染色证实为高级别发育异常的腺瘤性息肉，IHC证实了CLDN1的表达。
结论
CLDN1-IRDye800可通过超高分辨率荧光成像技术在CPC-APC小鼠中实现息肉可视化。小至1mm的息肉都可以可视化，这表明其在克服当前结肠息肉检测局限性方面具有临床潜力。
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