Department of Radiology, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, 519000, China.
Center for Interventional Medicine, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, 519000, China.
Eur J Nucl Med Mol Imaging. 2023 Jul;50(9):2787-2801. doi: 10.1007/s00259-023-06250-z. Epub 2023 May 5.
This study aimed to establish a near infrared fluorescent (NIRF) probe based on an EGFR&c-Met bispecific antibody for visualization of esophageal cancer (EC) and metastatic lymph nodes (mLNs).
EGFR and c-Met expression were assessed by immunohistochemistry. EGFR&c-Met bispecific antibody EMB01 was labeled with IRDye800cw. The binding of EMB01-IR800 was assessed by enzyme linked immunosorbent assay, flow cytometry, and immunofluorescence. Subcutaneous tumors, orthotopic tumors, and patient-derived xenograft (PDX) were established for in vivo fluorescent imaging. PDX models using lymph nodes with or without metastasis were constructed to assess the performance of EMB01-IR800 in differential diagnosis of lymph nodes.
The prevalence of overexpressing EGFR or c-Met was significantly higher than single marker either in EC or corresponding mLNs. The bispecific probe EMB01-IR800 was successfully synthesized, with strong binding affinity. EMB01-IR800 showed strong cellular binding to both Kyse30 (EGFR overexpressing) and OE33 (c-Met overexpressing) cells. In vivo fluorescent imaging showed prominent EMB01-IR800 uptake in either Kyse30 or OE33 subcutaneous tumors. Likewise, EMB01-IR800 exhibited superior tumor enrichment in both thoracic orthotopic esophageal squamous cell carcinoma and abdominal orthotopic esophageal adenocarcinoma models. Moreover, EMB01-IR800 produced significantly higher fluorescence in patient-derived mLNs than in benign lymph nodes.
This study demonstrated the complementary overexpression of EGFR and c-Met in EC. Compared to single-target probes, the EGFR&c-Met bispecific NIRF probe can efficiently depict heterogeneous esophageal tumors and mLNs, which greatly increased the sensitivity of tumor and mLN identification.
本研究旨在建立一种基于表皮生长因子受体（EGFR）和间质表皮转化因子（c-Met）双特异性抗体的近红外荧光（NIRF）探针，用于可视化食管癌（EC）和转移性淋巴结（mLNs）。
通过免疫组织化学评估 EGFR 和 c-Met 的表达。用 IRDye800cw 标记 EGFR 和 c-Met 双特异性抗体 EMB01。通过酶联免疫吸附试验、流式细胞术和免疫荧光评估 EMB01-IR800 的结合情况。建立皮下肿瘤、原位肿瘤和患者来源的异种移植（PDX）模型进行体内荧光成像。构建有或无转移的淋巴结 PDX 模型，以评估 EMB01-IR800 在淋巴结鉴别诊断中的性能。
在 EC 或相应的 mLNs 中，过表达 EGFR 或 c-Met 的患病率明显高于单一标志物。成功合成了具有强结合亲和力的双特异性探针 EMB01-IR800。EMB01-IR800 对 Kyse30（EGFR 过表达）和 OE33（c-Met 过表达）细胞均具有强烈的细胞结合。体内荧光成像显示，无论是 Kyse30 还是 OE33 皮下肿瘤，EMB01-IR800 均有明显的摄取。同样，EMB01-IR800 在胸内原位食管鳞状细胞癌和腹内原位食管腺癌模型中也表现出优越的肿瘤富集。此外，与良性淋巴结相比，EMB01-IR800 在患者衍生的 mLNs 中产生的荧光显著更高。
本研究证明了 EC 中 EGFR 和 c-Met 的互补过表达。与单靶点探针相比，EGFR 和 c-Met 双特异性 NIRF 探针能够有效地描绘出异质性食管肿瘤和 mLNs，大大提高了肿瘤和 mLN 识别的灵敏度。
