Thymidylate synthase disruption to limit cell proliferation in cell therapies

• DOI: 10.1016/j.ymthe.2024.06.014
• Publication Date: 2024-06-12T22:57:30Z
• Authors: Rocio Sartori-Mal...; Hossam Montaser; Inkeri Soppa; Solja Eurola; Juhana Juutila; Melanie Balaz; Henri Puttonen; Timo Otonkoski; Jonna Saarimäki-Vire; Kirmo Wartiovaara
• Keywords: Cell potency

Stem and progenitor cells hold great promise for regenerative medicine gene therapy approaches. However, transplantations of living entail a fundamental risk unwanted growth, potentially exacerbated by CRISPR-Cas9 or other genetic manipulations. Here, we describe safety system to control cell proliferation while allowing robust efficient manufacture, without any added elements. Inactivating TYMS, key nucleotide metabolism enzyme, in several lines resulted that proliferate only when supplemented with exogenous thymidine. Under supplementation, TYMS-/--pluripotent stem proliferate, produce teratomas successfully differentiate into therapeutic types such as pancreatic beta-cells. Our results suggest supplementation thymidine affects proliferation, but not the function cell-derived cells. After differentiation, postmitotic do require vitro vivo, shown production functional human insulin mice up 5 months after implantation stem-cell derived tissue.