Identification of a bladder cancer-specific ligand using a combinatorial chemistry approach.
Identification of a bladder cancer-specific ligand using a combinatorial chemistry approach.
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使用组合化学方法鉴定膀胱癌特异性配体。
DOI：
10.1016/j.urolonc.2010.06.011
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发表时间：
2012-09
期刊：
影响因子：
--
通讯作者：
Pan CX
中科院分区：
文献类型：
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作者：
Zhang H;Aina OH;Lam KS;de Vere White R;Evans C;Henderson P;Lara PN;Wang X;Bassuk JA;Pan CX
利用组合化学方法开发膀胱癌特异性配体。我们进行了高通量的一珠一化合物组合化学方法，以确定结合到膀胱移行细胞癌细胞的配体。全细胞结合试验允许成功鉴定选择性结合膀胱癌细胞的几种肽。来自临床膀胱癌标本和细胞系的单细胞悬浮液用于测定结合特异性。用小鼠异种移植物进行研究以确定配体之一的体内结合和靶向效率、特异性和生物分布。鉴定了一种名为PLZ 4（氨基酸序列：cQDGRMGFc）的环肽，其可以选择性地结合膀胱癌细胞系和来自人类患者的所有5种原代膀胱癌细胞，但不结合正常尿路上皮细胞、来自正常膀胱样本的细胞混合物、成纤维细胞和血细胞。PLZ 4与不同来源膀胱癌细胞系结合的比较显示，PLZ 4具有膀胱癌特异性（P0.05）。PLZ 4可以与用pH 6.0的尿液处理的肿瘤细胞结合，但不与从4名积极接受膀胱内卡介苗治疗的患者的尿液中收集的非癌细胞结合。体内和离体成像研究显示，通过尾静脉注射施用的与Cy5.5荧光染料连接的PLZ 4在从切除的新鲜人膀胱癌标本发育的小鼠异种移植物中特异性摄取。几种配体含有相同的DGR基序，但只有PLZ 4是膀胱癌特异性的。我们进行了丙氨酸步行和彩虹珠编码实验，发现C-末端GF残基对细胞结合也很重要，并调节结合特异性。PLZ 4有潜力用于非侵入性和晚期膀胱癌诊断和随访/监测期间的靶向治疗和成像检测。
To develop bladder cancer-specific ligands using a combinatorial chemistry approach. We performed a high-throughput one-bead one-compound combinatorial chemistry approach to identify ligands that bound to bladder transitional cell carcinoma cells. The whole-cell binding assay allowed successful identification of a few peptides that bound selectively to bladder cancer cells. Single cell suspensions derived from clinical bladder cancer specimens and cell lines were used to determine the binding specificity. Studies with mouse xenografts were performed to determine the in vivo binding and targeting efficiency, specificity, and biodistribution of one of the ligands. One cyclic peptide named PLZ4 (amino acid sequence: cQDGRMGFc) was identified that could selectively bind to bladder cancer cell lines and all of the 5 primary bladder cancer cells from human patients, but not to normal urothelial cells, cell mixtures from normal bladder specimens, fibroblasts, and blood cells. Comparison of PLZ4 binding to cell lines of different cancer origins showed that it was bladder cancer-specific (P _ 0.05). PLZ4 could bind to tumor cells treated with urine at pH 6.0, but not to noncancerous cells collected from the urine of 4 patients actively being treated with intravesical Bacillus Calmette-Guerin therapy. In vivo and ex vivo imaging studies showed that PLZ4 linked to Cy5.5 fluorescent dye administered via tail vein injection was specifically taken up in mouse xenografts developed from excised fresh human bladder cancer specimens. Several ligands contain the same DGR motif, but only PLZ4 was bladder cancer-specific. We performed alanine walk and rainbow bead coding experiments, and found that the C-terminal GF residues were also important for cell binding and modulated the binding specificity. PLZ4 has the potential to be used for targeted therapy and imaging detection during diagnosis and follow-up/surveillance of noninvasive and advanced bladder cancer.