Wnt signaling promotes hindgut fate commitment through regulating multi-lineage genes during hESC differentiation.
Wnt signaling plays essential roles in both embryonic pattern formation and postembryonic tissue homoestasis. High levels of Wnt activity repress foregut identity and facilitate hindgut fate through forming a gradient of Wnt signaling activity along the anterior-posterior axis. Here, we examined the mechanisms of Wnt signaling in hindgut development by differentiating human embryonic stem cells (hESCs) into the hindgut progenitors. We observed severe morphological changes when Wnt signaling was blocked by using Wnt antagonist Dkk1. We performed deep-transcriptome sequencing (RNA-seq) and identified 240 Wnt-activated genes and 2023 Wnt-repressed genes, respectively. Clusters of Wnt targets showed enrichment in specific biological functions, such as "gastrointestinal or skeletal development" in the Wnt-activated targets and "neural or immune system development" in the Wnt-repressed targets. Moreover, we adopted a high-throughput chromatin immunoprecipitation and deep sequencing (ChIP-seq) approach to identify the genomic regions through which Wnt-activated transcription factor TCF7L2 regulated transcription. We identified 83 Wnt direct target candidates, including the hindgut marker CDX2 and the genes relevant to morphogenesis (MSX1, MSX2, LEF1, T, PDGFRB etc.) through combinatorial analysis of the RNA-seq and ChIP-seq data. Together, our study identified a series of direct and indirect Wnt targets in hindgut differentiation, and uncovered the diverse mechanisms of Wnt signaling in regulating multi-lineage differentiation.
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China; University of Chinese Academy of Sciences, Beijing, China.
Xiujuan
X
Zhang
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Ying
Y
Chen
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Ying
Y
Ye
Core Genomic Facility, CAS Key Laboratory of Genome Sciences & Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China.
Jianfeng
J
Wang
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Hong
H
Wang
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Guohong
G
Yuan
State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Zhe
Z
Lin
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Yihui
Y
Wu
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Yan
Y
Zhang
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China; Division of Developmental Biology, Cincinnati Childrens Hospital Medical Center, Cincinnati, OH, United States; State Key Laboratory of Genetic Engineering, Institute of Genetics, Collaborative Innovation Center of Genetics and Development, School of Life Sciences, Fudan University, Shanghai, China. Electronic address: xinhua.lin@ioz.ac.cn.
Xinhua
X
Lin
D000075244:Apelin Receptors / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D004064:Digestive System / Q000166:cytology / Q000000981:diagnostic imaging* / Q000196:embryology*; D004707:Endoderm / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D063990:Gene Ontology; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D008648:Mesoderm / Q000378:metabolism; D009024:Morphogenesis / Q000235:genetics; D009940:Organoids / Q000254:growth & development / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D058118:Transcription Factor 7-Like 2 Protein / Q000378:metabolism; D014158:Transcription, Genetic; D060449:Wnt Signaling Pathway / Q000235:genetics
C104575:APLNR protein, human; D000075244:Apelin Receptors; D019718:Receptors, CXCR4; C544894:TCF7L2 protein, human; D058118:Transcription Factor 7-Like 2 Protein; D019009:Proto-Oncogene Proteins c-kit
APLNR; ChIP-seq; Hindgut specification; RNA-seq; Wnt targets; hESC
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