Identification of Cdca7 as a novel Notch transcriptional target involved in hematopoietic stem cell emergence

TY - JOUR

T1 - Identification of Cdca7 as a novel Notch transcriptional target involved in hematopoietic stem cell emergence

AU - Guiu, Jordi

AU - Bergen, Dylan J.M.

AU - de Pater, Emma

AU - Islam, Abul B.M.M.K.

AU - Ayllón, Verónica

AU - Gama-Norton, Leonor

AU - Ruiz-Herguido, Cristina

AU - González, Jessica

AU - López-Bigas, Nuria

AU - Menendez, Pablo

AU - Dzierzak, Elaine

AU - Espinosa, Lluis

AU - Bigas, Anna

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Hematopoietic stem cell (HSC) specification occurs in the embryonic aorta and requires Notch activation; however, most of the Notch-regulated elements controlling de novo HSC generation are still unknown. Here, we identify putative direct Notch targets in the aorta-gonad-mesonephros (AGM) embryonic tissue by chromatin precipitation using antibodies against the Notch partner RBPj. By ChIP-on-chip analysis of the precipitated DNA, we identified 701 promoter regions that were candidates to be regulated by Notch in the AGM. One of the most enriched regions corresponded to the Cdca7 gene, which was subsequently confirmed to recruit the RBPj factor but also Notch1 in AGM cells. We found that during embryonic hematopoietic development, expression of Cdca7 is restricted to the hematopoietic clusters of the aorta, and it is strongly up-regulated in the hemogenic population during human embryonic stem cell hematopoietic differentiation in a Notchdependent manner. Down-regulation of Cdca7 mRNA in cultured AGM cells significantly induces hematopoietic differentiation and loss of the progenitor population. Finally, using loss-of-function experiments in zebrafish, we demonstrate that CDCA7 contributes to HSC emergence in vivo during embryonic development. Thus, our study identifies Cdca7 as an evolutionary conserved Notch target involved in HSC emergence.

AB - Hematopoietic stem cell (HSC) specification occurs in the embryonic aorta and requires Notch activation; however, most of the Notch-regulated elements controlling de novo HSC generation are still unknown. Here, we identify putative direct Notch targets in the aorta-gonad-mesonephros (AGM) embryonic tissue by chromatin precipitation using antibodies against the Notch partner RBPj. By ChIP-on-chip analysis of the precipitated DNA, we identified 701 promoter regions that were candidates to be regulated by Notch in the AGM. One of the most enriched regions corresponded to the Cdca7 gene, which was subsequently confirmed to recruit the RBPj factor but also Notch1 in AGM cells. We found that during embryonic hematopoietic development, expression of Cdca7 is restricted to the hematopoietic clusters of the aorta, and it is strongly up-regulated in the hemogenic population during human embryonic stem cell hematopoietic differentiation in a Notchdependent manner. Down-regulation of Cdca7 mRNA in cultured AGM cells significantly induces hematopoietic differentiation and loss of the progenitor population. Finally, using loss-of-function experiments in zebrafish, we demonstrate that CDCA7 contributes to HSC emergence in vivo during embryonic development. Thus, our study identifies Cdca7 as an evolutionary conserved Notch target involved in HSC emergence.

UR - http://www.scopus.com/inward/record.url?scp=84911939394&partnerID=8YFLogxK

U2 - 10.1084/jem.20131857

DO - 10.1084/jem.20131857

M3 - Journal article

C2 - 25385755

AN - SCOPUS:84911939394

SN - 0022-1007

VL - 211

SP - 2411

EP - 2423

JO - Journal of Experimental Medicine

JF - Journal of Experimental Medicine

IS - 12

ER -