Inhibition of an NAD⁺ salvage pathway provides efficient and selective toxicity to human pluripotent stem cells

The tumorigenic potential of human pluripotent stem cells (hPSCs) is a major limitation to the widespread use of hPSC derivatives in the clinic. Here, we demonstrate that the small molecule STF-31 is effective at eliminating undifferentiated hPSCs across a broad range of cell culture conditions with important advantages over previously described methods that target metabolic processes. Although STF-31 was originally described as an inhibitor of glucose transporter 1, these data support the reclas-sificationofSTF-31asaspecific NAD⁺ salvage pathway inhibitor through the inhibition ofnicotinamide phosphoribosyltransferase (NAMPT). These findings demonstrate the importanceofanNAD⁺ salvage pathway in hPSC biology and describe how inhibition of NAMPT can effectively eliminate hPSCs from culture. These results will advance and accelerate the development of safe, clinically relevant hPSC- derived cell-based therapies.