Depletion of Demethylase KDM6 Enhances Early Neuroectoderm Commitment of Human PSCs

Abstract

Epigenetic modifications play a crucial role in neurogenesis, learning, and memory, but the study of their role in early neuroectoderm commitment from pluripotent inner cell mass is relatively lacking. Here we utilized the system of directed neuroectoderm differentiation from human embryonic stem cells and identified that KDM6B, an enzyme responsible to erase H3K27me3, was the most upregulated enzyme of histone methylation during neuroectoderm differentiation by transcriptome analysis. We then constructed KDM6B-null embryonic stem cells and found strikingly that the pluripotent stem cells with KDM6B knockout exhibited much higher neuroectoderm induction efficiency. Furthermore, we constructed a series of embryonic stem cell lines knocking out the other H3K27 demethylase KDM6A, and depleting both KDM6A and KDM6B, respectively. These cell lines together confirmed that KDM6 impeded early neuroectoderm commitment. By RNA-seq, we found that the expression levels of a panel of WNT genes were significantly affected upon depletion of KDM6. Importantly, the result that WNT agonist and antagonist could abolish the differential neuroectoderm induction due to manipulating KDM6 further demonstrated that WNT was the major downstream of KDM6 during early neural induction. Moreover, we found that the chemical GSK-J1, an inhibitor of KDM6, could enhance neuroectoderm induction from both embryonic stem cells and induced pluripotent stem cells. Taken together, our findings not only illustrated the important role of the histone methylation modifier KDM6 in early neurogenesis, providing insights into the precise epigenetic regulation in cell fate determination, but also showed that the inhibitor of KDM6 could facilitate neuroectoderm differentiation from human pluripotent stem cells.

KDM6B Negatively Regulates the Neurogenesis Potential of Apical Papilla Stem Cells via HES1

“…KDM6B, an enzyme that modifies histones, plays an essential role in early neurogenesis. Inhibition of its expression can promote neuroectodermal differentiation of human pluripotent stem cells [ 19 ]. In this study, we aim to analyze the biological role and mechanism of KDM6B in repairing neural tissue using SCAPs.…”

“…Furthermore, KDM6B is involved in regulating the development of medial, lateral, and preganglionic motor columns [ 18 ]. In the directed neuroectodermal differentiation system of human embryonic stem cells, it has been observed that the knockout of KDM6B leads to a higher efficiency of neuroectodermal induction in human pluripotent stem cells [ 19 ]. The origin of cells used in neuro-regenerative medicine is a matter of concern, with embryonic stem cells and neural stem cells being the ideal sources as they can serve as seed cells while shaping the neural regeneration microenvironment.…”

KDM6A facilitates Xist upregulation at the onset of X inactivation

“…Indeed, KDM6A mutations are associated with increased "stemness" in cancer and Kdm6a KO impairs mesoderm development in vivo largely through histone demethylase-independent mechanisms [20]. However, recent studies of Kdm6a KO models indicate that depletion of the enzyme does not block differentiation, and may even facilitate differentiation into neuroectoderm lineages [35,36]. Indeed, we only observed effects on germ layer marker genes after XCI has occurred and our KDM6A chromatin binding analyses suggest a direct effect of KDM6A on Xist expression during early XCI rather than an effect on differentiation itself.…”

Histone modifications in neurodifferentiation of embryonic stem cells

“…Meng et al. discovered that Inhibition of KDM6 (histone H3K27me3 demethylase) can improve neuroectoderm induction from both ES cells and iPS cells ( Meng et al., 2021 ). In addition, deletion of KMT2D (encoding histone H3K4 methyltransferase) may cause precocious iPSC-derived neuronal maturation due to transcriptional suppression of metabolic genes and proliferation defects ( Carosso et al., 2019 ).…”