拮抗恶性肿瘤转移靶向肽的筛选及时效性研究

Short peptide is a kind of ideal molecular for targeting tumor cells. Previously, we found a homing peptide TMTP1 which specifically bound to tumor micrometastases and TMVP1 which could target tumor lymphatics. And we established of a screening and verification system of targeted peptide. On this basis, we intend to analysis the repeat peptide sequences screened before by bioinformatic analysis to obtain the new peptide TMTP2 and TMVP2 and its transformation structure. We verify the specific affinity with tumor cells or lymphatics of TMTP2 and TMVP2 by the use of immunofluorescence and competitive antagonism experiments. We detect the effect of targeting recognition with tumor metastasis and lymphatics of TMTP2 and TMVP2 by a new small animal imaging in vivo by optimal structuring of tumor-bearing mouse tumor metastasis model. We explore the feasibility and timeliness of the peptide TMTP2, TMVP2 as peptide carrier by preparing conjugates of TMTP2 and TMVP2 with the appropriate effect clips. And then we search probable cell membrane receptor of the TMTP2 peptide using the Pull-down technology, find the membrane binding receptor is expected to be associated with tumor metastasis molecular markers. Finally, we complete the construction and complement of the screening and verification system of tumor targeted peptide in vivo. If the project is successful, it would provide us a practical way of tumor targeting peptide screening, and bring new opportunities and hope for the molecular targeted treatment of malignant tumor.

短肽是一种理想的肿瘤细胞靶向分子。前期我们首次筛查获得了肿瘤微小转移灶的靶向肽TMTP1及肿瘤淋巴管靶向肽TMVP1，并初步建立了靶向肽的验证体系。在此基础上，本项目拟通过生物信息学分析前期筛选获得的重复多肽序列，对新多肽TMTP2、TMVP2结构进行改进；利用免疫荧光法和竞争性拮抗实验，验证TMTP2、TMVP2与肿瘤细胞或淋巴管的特异性亲和力；优化构建荷瘤小鼠肿瘤转移模型，采用全新的小动物实体成像体内验证多肽对肿瘤转移灶及淋巴管的靶向性；选择合适的效应片段，探讨多肽TMTP2、TMVP2作为多肽载体的可行性与时效性；同时采用Pulldown技术寻找与TMTP2的受体，有望获得与肿瘤转移相关的分子标志，最终完成对恶性肿瘤靶向肽体内外筛查与验证体系的构建与补充。项目的顺利实施将为肿瘤靶向肽的筛查提供一种切实可行的途径，并为肿瘤的分子治疗带来新的契机和希望。

短肽是一种理想的肿瘤细胞靶向分子。前期我们首次筛查获得了肿瘤微小转移灶的靶向肽TMTP1及肿瘤淋巴管靶向肽TMVP1，并初步建立了靶向肽的验证体系。在此基础上，本项目拟通过生物信息学分析前期筛选获得的重复多肽序列，对新多肽TMTP2、TMVP2结构进行改进；利用免疫荧光法和竞争性拮抗实验，验证TMTP2、TMVP2与肿瘤细胞或淋巴管的特异性亲和力；优化构建荷瘤小鼠肿瘤转移模型，采用全新的小动物实体成像体内验证多肽对肿瘤转移灶及淋巴管的靶向性；选择合适的效应片段，探讨多肽TMTP2、TMVP2作为多肽载体的可行性与时效性；将筛选出的新多肽TMTP2、TMVP2标记放射性核素99mTc和68Ga，其能够对肿瘤组织显像；将新多肽连接ICG用于可肿瘤组织实时荧光显像；所获得的新型显像剂ICG-TMTP2,ICG-TMVP2为可视化荧光腹腔镜切除肿瘤提供了临床应用前景！