Department of Pediatrics , Emory University School of Medicine and Children's Healthcare of Atlanta , Atlanta , Georgia 30322 , United States.
School of Chemistry and Biochemistry , Georgia Institute of Technology , Atlanta , Georgia 30318 , United States.
ACS Chem Biol. 2018 Aug 17;13(8):2329-2338. doi: 10.1021/acschembio.8b00490. Epub 2018 Jul 19.
Sensitization to prodrugs via transgenic expression of suicide genes is a leading strategy for the selective elimination of potentially tumorigenic human pluripotent stem cells (hPSCs) in regenerative medicine, but transgenic modification poses safety risks such as deleterious mutagenesis. We describe here an alternative method of delivering suicide-inducing molecules explicitly to hPSCs using virus-like particles (VLPs) and demonstrate its use in eliminating undifferentiated hPSCs in vitro. VLPs were engineered from Qβ bacteriophage capsids to contain enhanced green fluorescent protein (EGFP) or cytosine deaminase (CD) and to simultaneously display multiple IgG-binding ZZ domains. After labeling with antibodies against the hPSC-specific surface glycan SSEA-5, EGFP-containing particles were shown to specifically bind undifferentiated cells in culture, and CD-containing particles were able to eliminate undifferentiated hPSCs with virtually no cytotoxicity to differentiated cells upon treatment with the prodrug 5-fluorocytosine.
通过转基因表达自杀基因使前药敏感化是再生医学中选择性消除潜在致瘤性人类多能干细胞（hPSC）的主要策略，但转基因修饰存在安全性风险，如有害的诱变。我们在这里描述了一种使用病毒样颗粒（VLPs）将自杀诱导分子专门递送到 hPSC 的替代方法，并证明其可用于体外消除未分化的 hPSC。VLPs 由 Qβ噬菌体衣壳工程改造而成，包含增强型绿色荧光蛋白（EGFP）或胞嘧啶脱氨酶（CD），并同时显示多个 IgG 结合 ZZ 结构域。用针对 hPSC 特异性表面糖蛋白 SSEA-5 的抗体标记后，发现含有 EGFP 的颗粒能够特异性结合培养中的未分化细胞，而含有 CD 的颗粒在使用前药 5-氟胞嘧啶处理时能够几乎无细胞毒性地消除未分化的 hPSC。
