GLIS3 Transcriptionally Activates WNT Genes to Promote Differentiation of Human Embryonic Stem Cells into 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 the upstream regulators of these morphogens remains incomplete. Here, we show that the Krüppel‐like zinc finger transcription factor GLI‐Similar 3 (GLIS3) can direct differentiation of human embryonic stem cells (hESCs) 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 Wingless/Integrated (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, and that inhibition of WNT signaling is sufficient to abrogate GLIS3‐induced posterior specification. Our findings suggest a potential role for GLIS3 in the regulation of A–P specification through direct transcriptional activation of WNT genes. Stem Cells2018StemCells2019;37:202–215 Expression of the Krüppel‐like zinc finger transcription factor GLI Similar 3 in human embryonic stem cells induces differentiation along the posterior neural progenitor lineage instead of the default anterior lineage through the transcriptional activation of the strong posteriorizing factor WNT3A.