Identification of novel regulators of developmental hematopoiesis using Endoglin regulatory elements as molecular probes

Nasrallah, R; Fast, EM; Solaimani, P; Knezevic, K; Eliades, A; Patel, R; Thambyrajah, R; Unnikrishnan, A; Thoms, J; Beck, D; Vink, CS; Smith, A; Wong, J; Shepherd, M; Kent, D; Roychoudhuri, R; Paul, F; Klippert, J; Hammes, A; Willnow, T; Göttgens, B; Dzierzak, E; Zon, LI; Lacaud, G; Kouskoff, V; Pimanda, JE

Identification of novel regulators of developmental hematopoiesis using Endoglin regulatory elements as molecular probes

Nasrallah, R Fast, EM Solaimani, P Knezevic, K Eliades, A Patel, R Thambyrajah, R Unnikrishnan, A Thoms, J Beck, D Vink, CS Smith, A Wong, J Shepherd, M Kent, D Roychoudhuri, R Paul, F Klippert, J Hammes, A Willnow, T Göttgens, B Dzierzak, E Zon, LI Lacaud, G Kouskoff, V Pimanda, JE

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Publication Type:: Journal Article
Citation:: Blood, 2016, 128 (15), pp. 1928 - 1939
Issue Date:: 2016-10-13

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Field	Value	Language
dc.contributor.author	Nasrallah, R	en_US
dc.contributor.author	Fast, EM	en_US
dc.contributor.author	Solaimani, P	en_US
dc.contributor.author	Knezevic, K	en_US
dc.contributor.author	Eliades, A	en_US
dc.contributor.author	Patel, R	en_US
dc.contributor.author	Thambyrajah, R	en_US
dc.contributor.author	Unnikrishnan, A	en_US
dc.contributor.author	Thoms, J	en_US
dc.contributor.author	Beck, D	en_US
dc.contributor.author	Vink, CS	en_US
dc.contributor.author	Smith, A	en_US
dc.contributor.author	Wong, J	en_US
dc.contributor.author	Shepherd, M	en_US
dc.contributor.author	Kent, D	en_US
dc.contributor.author	Roychoudhuri, R	en_US
dc.contributor.author	Paul, F	en_US
dc.contributor.author	Klippert, J	en_US
dc.contributor.author	Hammes, A	en_US
dc.contributor.author	Willnow, T	en_US
dc.contributor.author	Göttgens, B	en_US
dc.contributor.author	Dzierzak, E	en_US
dc.contributor.author	Zon, LI	en_US
dc.contributor.author	Lacaud, G	en_US
dc.contributor.author	Kouskoff, V	en_US
dc.contributor.author	Pimanda, JE	en_US
dc.date.available	2016-08-15	en_US
dc.date.issued	2016-10-13	en_US
dc.identifier.citation	Blood, 2016, 128 (15), pp. 1928 - 1939	en_US
dc.identifier.issn	0006-4971	en_US
dc.identifier.uri	http://hdl.handle.net/10453/116285
dc.description.abstract	© 2016 by The American Society of Hematology. Enhancers are the primary determinants of cell identity, and specific promoter/enhancer combinations of Endoglin (ENG) have been shown to target blood and endothelium in the embryo. Here, we generated a series of embryonic stem cell lines, each targeted with reporter constructs driven by specific promoter/enhancer combinations of ENG, to evaluate their discriminative potential and value as molecular probes of the corresponding transcriptome. The Eng promoter (P) in combination with the 28/17/19-kb enhancers, targeted cells in FLK1 mesoderm that were enriched for blast colony forming potential, whereas the P/28-kb enhancer targeted TIE21/c-KIT1/CD412 endothelial cells that were enriched for hematopoieticpotential. These fractions were isolated using reporter expression and their transcriptomes profiled by RNA-seq. There was high concordance between our signatures and those from embryos with defects at corresponding stages of hematopoiesis. Of the 6 genes that were upregulated in both hemogenic mesoderm and hemogenic endothelial fractions targeted by the reporters,LRP2, amultiligandreceptor,wasthe only gene that hadnot previously been associated with hematopoiesis.WeshowthatLRP2is indeedinvolved indefinitivehematopoiesisandbydoingsovalidatetheuseof reportergene-coupled enhancers as probes to gain insights into transcriptional changes that facilitate cell fate transitions.	en_US
dc.relation.ispartof	Blood	en_US
dc.relation.isbasedon	10.1182/blood-2016-02-697870	en_US
dc.subject.classification	Immunology	en_US
dc.subject.mesh	Cell Line	en_US
dc.subject.mesh	Endothelial Cells	en_US
dc.subject.mesh	Mesoderm	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Mice	en_US
dc.subject.mesh	Molecular Probes	en_US
dc.subject.mesh	Hematopoiesis	en_US
dc.subject.mesh	Embryo, Mammalian	en_US
dc.subject.mesh	Enhancer Elements, Genetic	en_US
dc.subject.mesh	Low Density Lipoprotein Receptor-Related Protein-2	en_US
dc.subject.mesh	Mouse Embryonic Stem Cells	en_US
dc.subject.mesh	Endoglin	en_US
dc.title	Identification of novel regulators of developmental hematopoiesis using Endoglin regulatory elements as molecular probes	en_US
dc.type	Journal Article
utslib.citation.volume	15	en_US
utslib.citation.volume	128	en_US
utslib.for	1103 Clinical Sciences	en_US
utslib.for	0903 Biomedical Engineering	en_US
utslib.for	1102 Cardiorespiratory Medicine and Haematology	en_US
utslib.for	1114 Paediatrics and Reproductive Medicine	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology/School of Biomedical Engineering
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology/School of Software
pubs.organisational-group	/University of Technology Sydney/Strength - AAI - Advanced Analytics Institute Research Centre
pubs.organisational-group	/University of Technology Sydney/Strength - CHT - Health Technologies
utslib.copyright.status	closed_access
pubs.issue	15	en_US
pubs.publication-status	Published	en_US
pubs.volume	128	en_US

Abstract:

© 2016 by The American Society of Hematology. Enhancers are the primary determinants of cell identity, and specific promoter/enhancer combinations of Endoglin (ENG) have been shown to target blood and endothelium in the embryo. Here, we generated a series of embryonic stem cell lines, each targeted with reporter constructs driven by specific promoter/enhancer combinations of ENG, to evaluate their discriminative potential and value as molecular probes of the corresponding transcriptome. The Eng promoter (P) in combination with the 28/17/19-kb enhancers, targeted cells in FLK1 mesoderm that were enriched for blast colony forming potential, whereas the P/28-kb enhancer targeted TIE21/c-KIT1/CD412 endothelial cells that were enriched for hematopoieticpotential. These fractions were isolated using reporter expression and their transcriptomes profiled by RNA-seq. There was high concordance between our signatures and those from embryos with defects at corresponding stages of hematopoiesis. Of the 6 genes that were upregulated in both hemogenic mesoderm and hemogenic endothelial fractions targeted by the reporters,LRP2, amultiligandreceptor,wasthe only gene that hadnot previously been associated with hematopoiesis.WeshowthatLRP2is indeedinvolved indefinitivehematopoiesisandbydoingsovalidatetheuseof reportergene-coupled enhancers as probes to gain insights into transcriptional changes that facilitate cell fate transitions.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/116285