Targeted Elimination of Tumorigenic Human Pluripotent Stem Cells Using Suicide-Inducing Virus-like Particles

• Type of Material: Conference Paper
• Authors: Antonio Rampoldi, Emory University; Stephen N Crooke, Georgia Institute of Technology; Marcela K Preininger, Emory University; Rajneesh Jha, Emory University; Joshua Maxwell, Emory University; Lingmei Ding, Emory University; Paul Spearman, Emory University; M.G. Finn, Georgia Institute of Technology; Chunhui Xu, Emory University
• Language: English
• Date: 2018-08-01
• Publisher: Emory University Libraries
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2018 American Chemical Society.
• Final Published Version (URL): http://dx.doi.org/10.1021/acschembio.8b00490
• Title of Journal or Parent Work: ACS Chemical Biology
• Conference or Event Name: STEEP Regenerative Engineering and Medicine (REM) Workshop
• Volume: 13
• Issue: 8
• Start Page: 2329
• End Page: 2338
• Grant/Funding Information: This project was supported in part by the NIH grant R21HL123928 to C.X, R01GM101421 to M.G.F., R01HL136345 to C.X., and a seed grant to C.X. and M.G.F. from the National Center for Advancing Translational Sciences of the National Institutes of Health under Award Number UL1TR000454.; S.N.C. was supported by a predoctoral research fellowship from the National Science Foundation.
• Keywords: DIFFERENTIATION; MOUSE; SELECTIVE ABLATION; CARDIOMYOCYTES; Biochemistry & Molecular Biology; GROWTH; BINDING DOMAIN; GENE; Science & Technology; Life Sciences & Biomedicine
• Research Categories: Chemistry, Biochemistry; Biology, Molecular; Engineering, Biomedical

Abstract

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.