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

Journal article (2016)

Authors

Rabab Nasrallah University of New South Wales, Australia
Eva M. Fast Harvard Medical School, Harvard University
Parham Solaimani Erasmus MC
Kathy Knezevic University of New South Wales, Australia
Alexia Eliades The University of Manchester
Rahima Patel The University of Manchester
Roshana Thambyrajah The University of Manchester
C.S. Vink The University of Edinburgh
John E. Pimanda University of New South Wales, Australia, Prince of Wales Hospital
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Published Date

2016

Language

English

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Abstract

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.