AMPK-p38 axis converts human pluripotent stem cells to naïve state

SUMMARY Pluripotent stem cells (PSCs) have been reported to exhibit two stages of pluripotency, ‘primed’ and ‘naïve’ states. Typical human PSCs (hPSCs) are in the primed state. Though several methods for conversion from primed to naïve state have been reported, the mechanism of the process is not fully understood. Here, we report that 5’ adenosine monophosphate-activated protein kinase (AMPK) and its downstream p38 is a signaling axis that can induce the naïve conversion of hPSCs with single pathway activation. The simple addition of an AMPK activator, 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR), or overexpression of a constitutive active form of p38 (CA-p38) alone in primed hPSCs induced naïve hPSCs that satisfied naïve state criteria: differentiation ability to three germ layers and naïve state-specific transcriptional expression, epigenomic resetting, and mitochondrial activity. RNA-seq analysis demonstrated that our AICAR- or CA-p38-induced naïve hPSCs show closely similar gene expression patterns to naïve state human embryonic stem cells (HNES1) derived from human inner cell mass (ICM). This novel and simple naïve conversion method provides new avenues for understanding and elucidating the fundamental mechanism of naïve conversion.