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

Rabab Nasrallah, Eva M. Fast, Parham Solaimani, Kathy Knezevic, Alexia Eliades, Rahima Patel, Roshana Thambyrajah, Ashwin Unnikrishnan, Julie Thoms, Dominik Beck, Chris S. Vink, Aileen Smith, Jason Wong, Mairi Shepherd, David Kent, Rahul Roychoudhuri, Fabian Paul, Julia Klippert, Annette Hammes, Thomas WillnowBertie Göttgens, Elaine Dzierzak, Leonard I. Zon, George Lacaud, Valerie Kouskoff, John E. Pimanda

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

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 -8/+7/+9kb enhancers, targeted cells in FLK1 mesoderm that were enriched for blast colony forming potential, whereas the P/-8kb enhancer targeted TIE2+/c-KIT+/CD41- endothelial cells that were enriched for hematopoietic potential. 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 six genes that were up-regulated in both hemogenic mesoderm and hemogenic endothelial fractions targeted by the reporters, LRP2, a multiligand receptor, was the only gene that had not previously been associated with hematopoiesis. We show that LRP2 is indeed involved in definitive hematopoiesis and by doing so validate the use of reporter gene coupled enhancers as probes to gain insights into transcriptional changes that facilitate cell fate transitions.

Original languageEnglish
Pages (from-to)1928-1939
Number of pages12
JournalBlood
Volume128
Issue number15
Early online date23 Aug 2016
DOIs
Publication statusPublished - 13 Oct 2016

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