Affinity peptide for targeted detection of dysplasia in Barrett's esophagus.
Affinity peptide for targeted detection of dysplasia in Barrett's esophagus.
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DOI：
10.1053/j.gastro.2010.07.007
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发表时间：
2010-11
期刊：
影响因子：
29.4
通讯作者：
Wang TD
中科院分区：
文献类型：
--
作者：
Li M;Anastassiades CP;Joshi B;Komarck CM;Piraka C;Elmunzer BJ;Turgeon DK;Johnson TD;Appelman H;Beer DG;Wang TD
异型增生是Barrett‘s食道的一种癌前状态，由于其扁平的结构和斑片状的分布，在筛查内窥镜下很难发现。多肽有望作为新型分子探针用于识别疾病特有的细胞表面靶标，并可被荧光标记进行检测。我们的目标是选择并验证一种与食道异型增生结合的亲和肽，用于未来的临床研究。通过使用Q-hTERT(肠化生)细胞去除非特异性结合并实现与OE33(食管腺癌)细胞的特异性结合，利用噬菌体展示进行多肽选择。结合噬菌体计数、酶联免疫吸附试验、流式细胞仪、竞争抑制和荧光显微镜均证实有选择性结合。在立体显微镜下，通过荧光强度与组织学的严格配准，以1 mm的间隔评价内窥镜下切除的标本上与异型增生结合的特异性多肽。筛选出SNFYMPL多肽序列，通过结合噬菌体计数、酶联免疫吸附试验和流式细胞仪检测，证明SNFYMPL与靶细胞有较好的结合。观察到与未标记多肽的竞争结合呈剂量依赖关系，测得Kd=164 nM，并在荧光显微镜下证实多肽与OE33细胞表面的结合。12例食管鳞状细胞癌、83例肠化生、61例异型增生和69例胃粘膜1 mm间隔荧光强度分别为46.5±1.6、62.3±5.8、100.0±9.0和42.4±3.0arb。SNFYMPL多肽序列可与Barrett‘s食管异型增生特异结合，并可在成像上被荧光标记为靶向癌前粘膜。
Dysplasia is a pre-malignant condition in Barrett's esophagus that is difficult to detect on screening endoscopy because of its flat architecture and patchy distribution. Peptides are promising for use as novel molecular probes that identify cell surface targets unique to disease, and can be fluorescence-labeled for detection. We aim to select and validate an affinity peptide that binds to esophageal dysplasia for future clinical studies. Peptide selection was performed using phage display by removing non-specific binders using Q-hTERT (intestinal metaplasia) cells and achieving specific binding against OE33 (esophageal adenocarcinoma) cells. Selective binding was confirmed on bound phage counts, ELISA, flow cytometry, competitive inhibition, and fluorescence microscopy. On stereomicroscopy, specific peptide binding to dysplasia on endoscopically resected specimens was assessed by rigorous registration of fluorescence intensity to histology in 1 mm intervals. The peptide sequence SNFYMPL was selected and demonstrated preferential binding to target cells on bound phage counts, ELISA, and flow cytometry. Reducing binding was observed on competition with unlabeled peptide in a dose dependent manner, an affinity of Kd = 164 nM was measured, and peptide binding to the surface of OE33 cells was validated on fluorescence microscopy. On esophageal specimens (n=12), the fluorescence intensity (mean±SEM) in 1 mm intervals classified histologically as squamous (n=145), intestinal metaplasia (n=83), dysplasia (n=61) and gastric mucosa (n=69) was 46.5±1.6, 62.3±5.8, 100.0±9.0, and 42.4±3.0 arb units, respectively. The peptide sequence SNFYMPL binds specifically to dysplasia in Barrett's esophagus, and can be fluorescence-labeled to target pre-malignant mucosa on imaging.
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