EVEN-SKIPPED HOMEOBOX 1 controls human ES cell differentiation by directly repressing GOOSECOID expression

Abstract

TGFß signaling patterns the primitive streak, yet little is known about transcriptional effectors that mediate the cell fate choices during streak-like development in mammalian embryos and in embryonic stem (ES) cells. Here we demonstrate that cross-antagonistic actions of EVEN-SKIPPED HOMEOBOX 1 (EVX1) and GOOSECOID (GSC) regulate cell fate decisions in streak-like progenitors derived from human ES cells exposed to BMP4 and/or activin. We found that EVX1 repressed GSC expression and promoted formation of posterior streak-like progeny in response to BMP4, and conversely that GSC repressed EVX1 expression and was required for development of anterior streak-like progeny in response to activin. Chromatin immunoprecipitation assays showed that EVX1 bound to the GSC 5'-flanking region in BMP4 treated human ES cells, and band shift assays identified two EVX1 binding sites in the GSC 5'-region. Significantly, we found that intact EVX1 binding sites were required for BMP4-mediated repression of GSC reporter constructs. We conclude that BMP4-induced EVX1 repress GSC directly and the two genes form the core of a gene regulatory network (GRN) controlling cell fates in streak-like human ES cell progeny.

DIGIT Is a Conserved Long Noncoding RNA that Regulates GSC Expression to Control Definitive Endoderm Differentiation of Embryonic Stem Cells

“…We used an shRNA to deplete GSC (Figure 4B) and determine how depletion of GSC affected differentiation of DE. Depletion of GSC expression inhibited SOX17 , as previously described (Kalisz et al, 2012) as well as FOXA2 and CXCR4 , which together mark DE. The results suggest that GSC is required for formation of DE during hESC differentiation.…”

“…Surprisingly, GSC −/− mice do not show a defect in gastrulation and survive to gestation with craniofacial defects (Rivera-Pérez et al, 1995). Loss of GSC expression in hESCs was previously show to be associated with significant reduction of SOX17 (Kalisz et al, 2012), which is expressed in both definitive and visceral endoderm (Shimoda et al, 2007). By quantifying the effects of GSC on additional genes, our results provide more convincing data that GSC is required for DE differentiation of hESCs.…”

Genome-Wide Temporal Profiling of Transcriptome and Open Chromatin of Early Cardiomyocyte Differentiation Derived From hiPSCs and hESCs

“…Many known TFs assigned to this module play important roles in mesoderm development, such as T , EOMES , MIXL1 , WNT3A , Dickkopf Wnt signaling pathway inhibitor 1 ( DKK1 ), Sp5 transcription factor ( SP5 ), mesogenin 1 ( MSGN1 ), even-skipped homeobox 1 ( EVX1 ) and caudal type homeobox 2 ( CDX2 ). 18-21, 27-31 We also observed that two transcripts which have not been well characterized before, AK127400 (coded by LOC100130256 ) and LOC440925 (known as “long intergenic non-protein coding RNA1124”, LINC01124 ), had highly-weighted connectivity within the module 4. Interestingly, LINC01124 and AK127400 are juxtaposed on chromosome 2 and surrounded by SP5 , MYO3B (myosin IIIB) and  GAD1 (glutamate decarboxylase 1) as a gene cluster (Online Figure V A), which were also assigned into the module 4.…”

Pilot Study on “Pericytic Mimicry” and Potential Embryonic/Stem Cell Properties of Angiotropic Melanoma Cells Interacting with the Abluminal Vascular Surface

“…Interestingly, from the coculture of EC and MC versus simultaneously MC alone and EC alone, 17 genes from the 27 known differentially expressed genes were linked to migratory and/or embryonic/cancer stem cell properties: nine overexpressed genes: CCL2, ICAM1, SELE, IL6, TRAF1, SERPINB2, CXCL6, PDGFB and EVX1, and eight underexpressed genes: CFDP1, BLID, RGNEF, MALT1, RANBP9, UPF1, PLAA and ZXDC. More specifically, these genes have demonstrated properties linked to (i) cancer cell migration: CCL2 [26,27], ICAM1 [28], IL6 [29], RGNEF [30] and RANBP9 [31]; (ii) cancer progression and metastasis: CCL2 [26,27,32], ICAM1 [33,34], SELE [35,36], TRAF1 [37], IL6 [29], SERPINB2 [38], CXCL6 [39], BLID [40], MALT1 [41], UPF1 [42], PLAA [43], RGNEF [44], ZXDC [45]; (iii) EMT: CCL2 [46] and IL6 [29] ; and (iv) embryonic and/or cancer stem cell properties: CCL2 [46][47][48] , PDGFB [49] , EVX1 [50], CFDP1 [51] and RANBP9 [52] . Furthermore, the 15 most significantly enriched functional groups analyzed by IPA included: development, cell movement, cancer, and embryonic development.…”