miR-200 and miR-96 families repress neural induction from human embryonic stem cells

Abstract

SUMMARYThe role of miRNAs in neuroectoderm specification is largely unknown. We screened miRNA profiles that are differentially changed when human embryonic stem cells (hESCs) were differentiated to neuroectodermal precursors (NEP), but not to epidermal (EPI) cells and found that two miRNA families, miR-200 and miR-96, were uniquely downregulated in the NEP cells. We confirmed zinc-finger E-box-binding homeobox (ZEB) transcription factors as a target of the miR-200 family members and identified paired box 6 (PAX6) transcription factor as the new target of miR-96 family members via gain-and loss-of-function analyses. Given the essential roles of ZEBs and PAX6 in neural induction, we propose a model by which miR-200 and miR-96 families coordinate to regulate neural induction.

ADAR1 is required for differentiation and neural induction by regulating microRNA processing in a catalytically independent manner

“…miR-96 and miR-200 families are known to suppress neural differentiation by targeting key neural transcription factors PAX6 and ZEB1 [36]. Consistent with their upregulation (Supplementary information, Table S5), we observed that pax6 and zeb1 mRNAs were significantly downregulated in ADAR1 KD H9 cells upon differentiation (data not shown).…”

“…At 10 and 17 days of neural differentiation, hESC-derived primitive and definitive neuroepthelial (NE) cells with scramble shRNA treatment presented typical columnar NE morphology and were organized into rosettes [36][37][38] (Figure 1D and 1E), similar to the structure of NE cells from WT hESC induction (as discussed below and data not shown). Noticeably, although differentiating hESCs with stable ADAR1 knockdown also formed rosette-like structures, these cells expressed low levels of PAX6, SOX1 and FOXG1 ( Figure 1D and Supplementary information, Table S1), and formed much Table S1.…”

“…As ADAR1 knockdown has little effect on pluripotency, we then proceeded to study the effect of ADAR1 deficiency on differentiation with a protocol summarized in Supplementary information, Figure S2A [36][37][38]. Briefly, hESCs were induced into the neural lineage by generating EBs followed by culturing in neural medium.…”

“…It is well known that self-renewal-related miR-302s promote ESC proliferation and suppress differentiation [45,46]; thus the delayed downregulation of miR-302s upon ADAR1 depletion could lead to the suppression of hESC differentiation. Furthermore, some miRNAs, such as miR-96 and miR-200s, that repress neural induction [36], were also upregulated in ADAR1 KD samples (Supplementary information, Table S5). This is in agreement with the notion that ADAR1 knockdown resulted in delayed neural induction (Figure 1).…”

MicroRNAs in Endothelial Development and Differentiation

“…Finally, in this study we have demonstrated for the first time that miRNA-200c-ZEB1 axis is a critical regulator for chick embryonic blood vessel formation by in vivo inhibition of miRNA-200C or −150 in developing chick embryos. In the meantime, other researchers also reported that miR-200c promotes mesodermal specification while repressing neuroectodermal differentiation from ESCs [95], suggesting that miR-200c may play a cell-autonomous role in mediating EC differentiation from stem cells. Interestingly, miR-200b displays an antiangiogenic activity in tumor and in embryonic vascular development [96,97], partially through repressing the crucial endothelial lineage related transcription factor E26 oncogene homolog 1 (Ets-1) [98].…”