GSE109562
GLIS3 Transcriptionally Activates WNT Genes to Promote Differentiation of Human Embryonic Stem Cells to Posterior Neural Progenitors
Anterior-posterior (A-P) specification of the neural tube involves initial acquisition of anterior fate followed by the induction of posterior characteristics in the primitive anterior neuroectoderm. Several morphogens have been implicated in the regulation of A-P neural patterning; however, our understanding of factors regulating these morphogens remains incomplete. Here we show that the Krüppel-like zinc finger transcription factor GLI-Similar 3 (GLIS3) directs differentiation of human embryonic stem cells into posterior neural progenitor cells in lieu of the default anterior pathway. Transcriptomic analyses reveal that this switch in cell fate is due to rapid activation of an autocrine WNT signaling pathway. Mechanistically, through genome-wide RNA-Seq, ChIP-Seq and functional analyses, we show that GLIS3 binds to and directly regulates the transcription of several WNT genes, including the strong posteriorizing factor WNT3A. Inhibition of WNT signaling is sufficient to abrogate GLIS3-induced posterior specification. Altogether, our findings suggest a critical role for GLIS3 in A-P specification through the direct transcriptional activation of WNT genes. Overall design: We performed ChIP-Seq analysis to map genome-wide occupancy of exogenous, HA-tagged GLIS3 in G3-hESCs(+) after 12 h of NPC induction and employed RNA-Seq to profile global gene expression changes at various time points during G3-hESC to NPC differentiation (0 h, 12 h, 1, 2, 3, and 7 days), in the presence or absence of DOX.