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

Jordi Guiu; Dylan J.M. Bergen; Emma de Pater; Abul B.M.M.K. Islam; Verónica Ayllón; Leonor Gama-Norton; Cristina Ruiz-Herguido; Jessica González; Nuria López-Bigas; Pablo Menendez; Elaine Dzierzak; Lluis Espinosa; Anna Bigas

doi:10.1084/jem.20131857

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

Jordi Guiu, Dylan J.M. Bergen, Emma de Pater, Abul B.M.M.K. Islam, Verónica Ayllón, Leonor Gama-Norton, Cristina Ruiz-Herguido, Jessica González, Nuria López-Bigas, Pablo Menendez, Elaine Dzierzak, Lluis Espinosa, Anna Bigas^*

^*Corresponding author for this work

33 Citations (Scopus)

Abstract

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.

Original language	English
Journal	Journal of Experimental Medicine
Volume	211
Issue number	12
Pages (from-to)	2411-2423
Number of pages	13
ISSN	0022-1007
DOIs	https://doi.org/10.1084/jem.20131857
Publication status	Published - 1 Jan 2014
Externally published	Yes

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Guiu, J., Bergen, D. J. M., de Pater, E., Islam, A. B. M. M. K., Ayllón, V., Gama-Norton, L., Ruiz-Herguido, C., González, J., López-Bigas, N., Menendez, P., Dzierzak, E., Espinosa, L., & Bigas, A. (2014). Identification of Cdca7 as a novel Notch transcriptional target involved in hematopoietic stem cell emergence. Journal of Experimental Medicine, 211(12), 2411-2423. https://doi.org/10.1084/jem.20131857

Guiu, J, Bergen, DJM, de Pater, E, Islam, ABMMK, Ayllón, V, Gama-Norton, L, Ruiz-Herguido, C, González, J, López-Bigas, N, Menendez, P, Dzierzak, E, Espinosa, L & Bigas, A 2014, 'Identification of Cdca7 as a novel Notch transcriptional target involved in hematopoietic stem cell emergence', Journal of Experimental Medicine, vol. 211, no. 12, pp. 2411-2423. https://doi.org/10.1084/jem.20131857

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title = "Identification of Cdca7 as a novel Notch transcriptional target involved in hematopoietic stem cell emergence",

abstract = "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.",

author = "Jordi Guiu and Bergen, {Dylan J.M.} and {de Pater}, Emma and Islam, {Abul B.M.M.K.} and Ver{\'o}nica Ayll{\'o}n and Leonor Gama-Norton and Cristina Ruiz-Herguido and Jessica Gonz{\'a}lez and Nuria L{\'o}pez-Bigas and Pablo Menendez and Elaine Dzierzak and Lluis Espinosa and Anna Bigas",

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doi = "10.1084/jem.20131857",

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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.

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DO - 10.1084/jem.20131857

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SP - 2411

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

Abstract

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