Phage display-derived peptides for osteosarcoma imaging.
Phage display-derived peptides for osteosarcoma imaging.
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DOI：
10.1158/1078-0432.ccr-10-0968
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发表时间：
2010-08-15
期刊：
影响因子：
--
通讯作者：
Chen X
中科院分区：
文献类型：
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作者：
Sun X;Niu G;Yan Y;Yang M;Chen K;Ma Y;Chan N;Shen B;Chen X
骨肉瘤是儿童期最常见的原发性恶性骨肿瘤，然而，其生存率在过去20年中保持不变。为了改善现有的诊断和治疗方法，并扩大可用于早期检测和评估肿瘤对治疗的反应的成像剂的范围，我们进行了基于噬菌体展示的筛选，以筛选特异性结合骨肉瘤细胞的肽序列。从包含2.7 ×109个不同展示肽的Ph.D.™-12噬菌体展示肽文库中，在143 B骨肉瘤肿瘤细胞上进行4轮体外选择后富集了一种肽，其中293 T人胚肾细胞作为对照。将肽和展示肽的噬菌体克隆与荧光染料缀合，用于体外细胞和离体肿瘤组织染色。用18F进一步标记肽用于正电子发射断层扫描（PET）成像研究。用18F标记的骨肉瘤特异性肽进行细胞摄取和流出以及离体生物分布。ASGALSPSRLDT是从生物淘选中分离到的优势序列，命名为OSP-1。OSP-1与肝素酶II/III家族蛋白具有显著的同源性，其与硫酸乙酰肝素蛋白聚糖（HSPG）结合并反应。荧光染色显示FITC-OSP-1-噬菌体或Cy5.5-OSP-1与一组骨肉瘤细胞系具有高结合，与UM-SCC 1人头颈部鳞状细胞癌细胞的结合少得多，并且几乎不与293 T细胞结合;而乱序肽OSP-S几乎不与所有细胞系结合。在体外和体内，18F-OSP-1在143 B肿瘤细胞中的蓄积均显著高于18F-OSP-S。18F-OSP-1在143 B肿瘤中的摄取也高于UM-SCC-1肿瘤。我们的数据表明，OSP-1肽是骨肉瘤特异性的，OSP-1的结合位点可能与硫酸乙酰肝素蛋白多糖有关。适当标记的OSP-1肽有可能作为骨肉瘤显像的新探针。
Osteosarcoma represents the most common malignant primary bone tumor in childhood; however, the survival rate has remained unchanged for past 20 years. To improve existing diagnosis and treatment methods and broaden the spectrum of imaging agents that can be used for early detection and assessment of tumor response to therapy, we performed a phage display-based screening for peptide sequences that bind specifically to osteosarcoma cells. From the Ph.D.™ −12 phage display peptide library comprising 2.7 ×109 different displayed peptides, one peptide was enriched after 4 rounds of in vitro selection on 143B osteosarcoma tumor cells with 293T human embryonic kidney cells as a control. Both the peptide and the phage clone displaying the peptide were conjugated with fluorescent dyes for in vitro cell and ex vivo tumor tissue staining. The peptide was further labeled with 18F for positron emission tomography (PET) imaging studies. Cell uptake and efflux and ex vivo biodistribution were also performed with 18F labeled osteosarcoma specific peptide. ASGALSPSRLDT was the dominant sequence isolated from biopanning and named as OSP-1. OSP-1 shares a significant homology with heparinase II/III family protein, which binds and reacts with heparan sulfate proteoglycans (HSPGs). The fluorescence staining showed that FITC-OSP-1-phage or Cy5.5-OSP-1 had high binding with a panel of osteosarcoma cell lines, much less binding with UM-SCC1 human head and neck squamous cell carcinoma cells, and almost no binding with 293T cells; whereas the scrambled peptide OSP-S had virtually no binding to all the cell lines. 18F-OSP-1 had significantly higher accumulation in 143B tumor cells both in vitro and in vivo than 18F-OSP-S. 18F-OSP-1 also had higher uptake in 143B tumors than UM-SCC-1 tumors. Our data suggest that OSP-1 peptide is osteosarcoma specific, and the binding site of OSP-1 might be related to heparan sulfate proteoglycans. Appropriately labeled OSP-1 peptide has the potential to serve as a novel probe for osteosarcoma imaging.