Mouse Embryonic Stem Cell Pluripotency Factors Regulate RNA Methylation

The pluripotency of embryonic stem cells (ESCs) is actively promoted by a diverse set of factors, including leukemia inhibitory factor (LIF), glycogen synthase kinase-3 (Gsk-3) and mitogen-activated protein kinase kinase (MEK) inhibitors, ascorbic acid, and α-ketoglutarate. Strikingly, several of these factors intersect with the post-transcriptional methylation of RNA (m ⁶ A), which has also been shown to play a role in ESC pluripotency. Therefore, we explored the possibility that these factors converge on this biochemical pathway to promote the retention of ESC pluripotency. Mouse ESCs were treated with various combinations of small molecules, and the relative levels of m ⁶ A RNA were measured, as well as the expression of genes marking naïve and primed ESCs. The most surprising result was the discovery that replacing glucose with high levels of fructose pushed ESCs to a more naïve state and reduced m ⁶ A RNA abundance. Our results suggest a correlation between molecules previously shown to promote the retention of ESC pluripotency and m ⁶ A RNA levels, strengthening a molecular connection between reduced m ⁶ A RNA and the pluripotent state, and provides a foundation for future mechanistic studies on the role of m ⁶ A and ESC pluripotency.