Lactate and lactylation of RNA splicing factors triggered metabolic code reverse central dogma modulates totipotency development

Totipotent cells mark the inception of mammalian development and are present in the body for an extremely brief duration. Exploring the regulation of stem cell totipotency would help us to understand the mammalian embryo development and accelerate its application on regeneration medicine. Here using high responsive lactate probe-Fila, we revealed that high lactate signal was related to stem cell totipotency, and supplement of lactate captured totipotent stem cell from naïve or primed stem cells significantly both in human and mouse. Then, we discovered that the high Kla intensity was a character of totipotency in embryo cells and cell lines, and the Kla intensity decreased following the totipotency development. Moreover, using high throughput lactylome and proteome analysis, we found the highest Kla intensity and sites in totipotent stem cells both on the cellular and subcellular. Notably, we revealed that Kla proteins participated in almost fundamental biological pathways such as RNA splicing, DNA repair and chromatin organization et al., and the Kla proteins related to these pathways represented a dynamic change during the totipotency development. We further focused on the most enrichment pathway RNA splicing, and revealed that the high Kla level on RNA splicing factors inhibited the splicing efficiency to activate the totipotent genes and silence the pluripotent genes, respectively. Finally, we screened and identified the Kla on six critical RNA splicing factors were essential for totipotency. Our work highlights lactate as a master and conservative metabolite, and reveals “lactate triggered-metabolic codon reversed central dogma” that involves the lactate driven Kla on RNA splicing factors to regulate the pre-mRNA splicing efficiency in the context of totipotency development.