Identification of Tumor Specific Peptide as EpCAM Ligand and Its Potential Diagnostic and Therapeutic Clinical Application
Identification of Tumor Specific Peptide as EpCAM Ligand and Its Potential Diagnostic and Therapeutic Clinical Application
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作为 EpCAM 配体的肿瘤特异性肽的鉴定及其潜在的诊断和治疗临床应用
DOI：
10.1021/acs.molpharmaceut.9b00185
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发表时间：
2019-05-01
影响因子：
4.9
通讯作者：
Li, Meng
中科院分区：
文献类型：
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作者：
Ma, Xiaowen;Kang, Xiaoyu;Li, Meng
正在开发用于早期肿瘤检测和治疗的肿瘤靶向剂。我们以前确定了肽SNFYMPL（SNF*），并证明了其特异性结合到人类食管标本的高度异型增生（HGD）和腺癌与成像离体。在这里，我们的目标是确定这种肽的目标，并研究其在成像和药物输送中的潜在应用。通过SNF* 结合亲和层析、质谱、Western blot、酶联免疫吸附试验（ELISA）和分子对接等方法，我们发现上皮细胞粘附分子（EpCAM）是SNF* 的潜在靶点。接下来，我们发现FITC标记的SNF*（SNF*-FITC）与EpCAM抗体共定位在食管腺癌细胞OE 33的表面上，并且SNF*-FITC结合模式在EpCAM敲低或外源性EpCAM转染后显著改变。根据TCGA的数据，我们证明EpCAM在17种癌症中过表达。以结肠和胃腺癌细胞和组织为例，我们发现SNF*-FITC结合的模式与EpCAM抗体共定位，并且SNF* 结合不上调EpCAM下游Wnt信号。随后，我们将SNF* 与我们先前构建的聚（组氨酸）-PEG/DSPE共聚物胶束缀合。SNF* 标记可显著提高胶束与结肠癌和胃腺癌细胞的体外结合能力，增强胶束的体内抗肿瘤作用，降低胶束的毒性。总之，我们鉴定并验证了SNF* 作为EpCAM的特异性肽。证实了SNF* 肽在多种肿瘤监测和肿瘤靶向治疗中的未来潜在用途。
Tumor targeting agents are being developed for early tumor detection and therapeutics. We previously identified the peptide SNFYMPL (SNF*) and demonstrated its specific binding to human esophageal specimens of high-grade dysplasia (HGD) and adenocarcinoma with imaging ex vivo. Here, we aim to identify the target for this peptide and investigate its potential applications in imaging and drug delivery. With SNF* conjugated affinity chromatography, mass spectrum, Western blot, enzyme-linked immunosorbent assay (ELISA), and molecular docking, we found that the epithelial cell adhesion molecule (EpCAM) was the potential target of SNF*. Next, we showed that FITC-labeled SNF* (SNF*-FITC) colocalized with EpCAM antibody on the surface of esophageal adenocarcinoma cells OE33, and SNF*-FITC binding patterns significantly changed after EpCAM knockdown or exogenous EpCAM transfection. With the data from TCGA, we demonstrated that EpCAM was overexpressed in 17 types of cancers. Using colon and gastric adenocarcinoma cells and tissues as examples, we found that SNF*-FITC bound in a pattern was colocalized with EpCAM antibody, and the SNF* binding did not upregulate the EpCAM downstream Wnt signals. Subsequently, we conjugated SNF* with our previously constructed poly(histidine)-PEG/DSPE copolymer micelles. SNF* labeling significantly improved the micelle binding with colon and gastric adenocarcinoma cells in vitro, and enhanced the antitumor effects and decreased the toxicities of the micelles in vivo. In conclusion, we identified and validated SNF* as a specific peptide for EpCAM. The future potential use of SNF* peptide in multiple tumor surveillance and tumor-targeted therapeutics was demonstrated.