Thymidylate synthase disruption to limit cell proliferation in cell therapies
11832 Microbiology and virology
Pluripotent Stem Cells
Medical biotechnology
Cell- and Tissue-Based Therapy
Cell Differentiation
Thymidylate Synthase
Cell Line
Mice
Insulin-Secreting Cells
Humans
Animals
Original Article
CRISPR-Cas Systems
Cell Proliferation
Thymidine
DOI:
10.1016/j.ymthe.2024.06.014
Publication Date:
2024-06-12T22:57:30Z
AUTHORS (10)
ABSTRACT
Stem and progenitor cells hold great promise for regenerative medicine and gene therapy approaches. However, transplantation of living cells entails a fundamental risk of unwanted growth, potentially exacerbated by CRISPR-Cas9 or other genetic manipulations. Here, we describe a safety system to control cell proliferation while allowing robust and efficient cell manufacture, without any added genetic elements. Inactivating TYMS, a key nucleotide metabolism enzyme, in several cell lines resulted in cells that proliferate only when supplemented with exogenous thymidine. Under supplementation, TYMS-/--pluripotent stem cells proliferate, produce teratomas, and successfully differentiate into potentially therapeutic cell types such as pancreatic β cells. Our results suggest that supplementation with exogenous thymidine affects stem cell proliferation, but not the function of stem cell-derived cells. After differentiation, postmitotic cells do not require thymidine in vitro or in vivo, as shown by the production of functional human insulin in mice up to 5 months after implantation of stem cell-derived pancreatic tissue.
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