Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology, Tsukuba Central 2, 1-1-1 Umezono, Tsukuba, Ibaraki, 305-8568, Japan.
Tsukuba Advanced Research Alliance, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8577, Japan.
Chembiochem. 2019 Jun 14;20(12):1606-1611. doi: 10.1002/cbic.201900086. Epub 2019 Apr 18.
Human pluripotent stem cells (hPSCs) are attractive resources for regenerative medicine, but medical applications are hindered by their tumorigenic potential. Previously, a hPSC-specific lectin probe, rBC2LCN, was identified through comprehensive glycome analysis by using high-density lectin microarrays. Herein, a lectin-doxorubicin (DOX) prodrug conjugate, with controllable photolysis activation for the elimination of tumorigenic human induced pluripotent stem cells, has been developed. rBC2LCN was fused with a biotin-binding protein, tamavidin (BC2Tama), and the fusion protein was expressed in Escherichia coli and purified by means of affinity chromatography. BC2Tama was then conjugated with doxorubicin-photocleavable biotin (DOXPCB). The BC2Tama-DOXPCB conjugates were observed to bind to hPSCs followed by internalization. Upon exposure to ultraviolet light, DOX was released inside the cells, which allowed specific killing of the hPSCs. Thus, BC2Tama-DOXPCB should be useful for the targeted elimination of hPSCs contained in hPSC-derived cell therapy products. This is the first report of the generation of lectin-prodrug conjugates. BC2Tama should be applicable for the targeted delivery of various types of biotinylated compounds into hPSCs.
人多能干细胞（hPSCs）是再生医学的有吸引力的资源，但它们的致瘤潜力阻碍了它们的医学应用。先前，通过使用高密度凝集素微阵列进行全面糖组分析，鉴定了一种 hPSC 特异性凝集素探针 rBC2LCN。在此，开发了一种凝集素-阿霉素（DOX）前药缀合物，具有可控的光解激活，用于消除致瘤性人诱导多能干细胞。rBC2LCN 与生物素结合蛋白，链霉亲和素（BC2Tama）融合，并通过亲和层析在大肠杆菌中表达和纯化。然后将 BC2Tama 与阿霉素光裂解生物素（DOXPCB）缀合。观察到 BC2Tama-DOXPCB 缀合物与 hPSCs 结合后内化。暴露于紫外线下，细胞内释放 DOX，从而特异性杀死 hPSCs。因此，BC2Tama-DOXPCB 应该可用于靶向消除 hPSC 衍生细胞治疗产品中包含的 hPSCs。这是生成凝集素-前药缀合物的第一个报告。BC2Tama 应该适用于将各种类型的生物素化化合物靶向递送至 hPSCs。
