miR-335 promotes mesendodermal lineage segregation and shapes a transcription factor gradient in the endoderm.
Transcription factors (TFs) pattern developing tissues and determine cell fates; however, how spatio-temporal TF gradients are generated is ill defined. Here we show that miR-335 fine-tunes TF gradients in the endoderm and promotes mesendodermal lineage segregation. Initially, we identified miR-335 as a regulated intronic miRNA in differentiating embryonic stem cells (ESCs). miR-335 is encoded in the mesoderm-specific transcript (Mest) and targets the 3'-UTRs of the endoderm-determining TFs Foxa2 and Sox17. Mest and miR-335 are co-expressed and highly accumulate in the mesoderm, but are transiently expressed in endoderm progenitors. Overexpression of miR-335 does not affect initial mesendoderm induction, but blocks Foxa2- and Sox17-mediated endoderm differentiation in ESCs and ESC-derived embryos. Conversely, inhibition of miR-335 activity leads to increased Foxa2 and Sox17 protein accumulation and endoderm formation. Mathematical modeling predicts that transient miR-335 expression in endoderm progenitors shapes a TF gradient in the endoderm, which we confirm by functional studies in vivo. Taken together, our results suggest that miR-335 targets endoderm TFs for spatio-temporal gradient formation in the endoderm and to stabilize lineage decisions during mesendoderm formation.
Institute of Stem Cell Research, Helmholtz Zentrum München, D-85764 Neuherberg, Germany.
Dapeng
D
Yang
Dominik
D
Lutter
Ingo
I
Burtscher
Lena
L
Uetzmann
Fabian J
FJ
Theis
Heiko
H
Lickert
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics*; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D004707:Endoderm / Q000166:cytology* / Q000196:embryology / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D024002:HMGB Proteins / Q000235:genetics / Q000378:metabolism; D051542:Hepatocyte Nuclear Factor 3-beta / Q000235:genetics / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D055760:SOXF Transcription Factors / Q000235:genetics / Q000378:metabolism; D062211:Spatio-Temporal Analysis; D014157:Transcription Factors / Q000378:metabolism*
C496503:Foxa2 protein, mouse; D024002:HMGB Proteins; D035683:MicroRNAs; C541986:Mirn335 microRNA, mouse; D055760:SOXF Transcription Factors; C511606:Sox17 protein, mouse; D014157:Transcription Factors; D051542:Hepatocyte Nuclear Factor 3-beta
Endoderm; Foxa2; Gastrulation; Mesendoderm; Mir335; Mouse; Sox17; miR-335