Identification of a Novel ITGαvβ6-Binding Peptide Using Protein Separation and Phage Display
Identification of a Novel ITGαvβ6-Binding Peptide Using Protein Separation and Phage Display
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DOI：
10.1158/1078-0432.ccr-16-3217
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发表时间：
2017-08-01
影响因子：
11.5
通讯作者：
Haberkorn, Uwe
中科院分区：
文献类型：
--
作者：
Altmann, Annette;Sauter, Max;Haberkorn, Uwe
目的：实验设计：为了筛选肿瘤特异性肽，采用ProteomeLab PF 2D系统分离HNO 97肿瘤细胞膜蛋白质组和相应的HNO 97细胞膜蛋白质组，利用向日葵胰蛋白酶噬菌体展示文库（SFTI 8 Ph）进行交替生物淘选。分别采用放射性HPLC、结合实验和表面等离子体共振分析（SPR）对新候选物的稳定性、结合特性和亲和力进行了体外评估。结果：我们鉴定了一种新的ITG α（v）β（6）结合肽（SFITGv 6），其氨基酸序列为FRGDLMQL。SFITGv 6在24小时内保持稳定，并对ITG α v β具有高亲和力（KD = 14.8 nmol/L）（6）。在HNO 97细胞中，测量到的最大摄取和内化分别高达37.3%和37.5%。HNO 97异种移植Balb/c nu/nu小鼠的小动物PET成像和生物分布研究显示，在注射后30至60分钟，Ga-68和Lu-177标记的DOTA-SFITGv 6分别发生肿瘤特异性蓄积。此外，肽组化显示生物素标记的SFITGv 6与HNSCC肿瘤以及乳腺癌和肺癌来源的脑转移瘤的强且均匀的结合。最后，HNSCC和NSCLC患者的第一次PET/CT扫描显示SFITGv 6特异性地在肿瘤中积累，但不在炎性lesions.Conclusions：因此，SFITGv 6代表了一种新的强大的示踪剂，用于成像，并可能用于ITG α（v）β（6）阳性癌的腔内放射治疗。(C)2017年AACR。
Purpose: Targeted therapies are regarded as promising approaches to increase 5-year survival rate of head and neck squamous cell carcinoma (HNSCC) patients.Experimental design: For the selection of carcinoma-specific peptides membrane proteome of HNO97 tumor cells fractionated by the ProteomeLab PF2D system and corresponding HNO97 cells were deployed for an alternating biopanning using a sunflower trypsin inhibitor1-based phage display (SFTI8Ph) library. Stability, binding properties and affinity of novel candidates were assessed in vitro using radio-HPLC, binding experiments and surface plasmon resonance assay (SPR), respectively. Subsequently, the affinity of the peptide was verified in situ by using peptide histochemistry, in vitro using flow cytometry, and in vivo by positron emissions tomography (PET/CT).Results: We identified a novel ITG alpha(v)beta(6) binding peptide (SFITGv6) containing the amino acid sequence FRGDLMQL. SFITGv6 provides stability over a period of 24 hours and demonstrates high affinity (K-D = 14.8 nmol/L) for ITG alpha v beta(6). In HNO97 cells, a maximal uptake and internalization of up to 37.3% and 37.5%, respectively, was measured. Small-animal PET imaging and biodistribution studies of HNO97 xenografted Balb/c nu/nu mice showed tumor-specific accumulation of Ga-68- and Lu-177-labeled DOTA-SFITGv6, respectively, 30 to 60 minutes after injection. Moreover, peptide histochemistry revealed a strong and homogenous binding of biotin-labeled SFITGv6 to HNSCC tumors and breast-and lung cancer-derived brain metastases. Finally, first PET/CT scans of HNSCC and NSCLC patients displayed SFITGv6 accumulation specifically in tumors, but not in inflammatory lesions.Conclusions: Thus, SFITGv6 represents a novel powerful tracer for imaging and possibly for endoradiotherapy of ITG alpha(v)beta(6)-positive carcinoma. (C) 2017 AACR.