Differentiation of Eng-/- Embryonic Stem Cells in Embryoid Bodies, a Paradigm for Vascuaogenesis/ Angiogenesis. Defects in HHT and a Potential Drug Screen

 

Treatment of Disease $50,000 Research Grant Award

Professor Christine Mummery
Utrecht, The Netherlands

 

 

 

This study will examine the effects of Thalidomide on blood vessel structure. According to Professor Mummery, “By studying the effects of thalidomide on vessel stabilization, we propose (1) to validate the Eng-/vasculogenesis/angiogenesis assay as a useful method in screening potential HHT drugs, particularly those directed towards improving vessel robustness and (2) to determine the mechanisms underlying vessel stabilization. We will attempt to generate a human model of HHT development in vitro.”

Research Plan

  1. Mechanism underlying vessel stabilization. By using FACS sorting, ECs and SMCs will be isolated from 18 day Eng-/- EBs treated or otherwise with thalidomide. Transcriptional profiles will be compared both by microarray analysis and by real time RT-PCR to identify signaling pathways that may underlie the effects observed.
  2. Validation of the EB vasculogenesis/angiogenesis assay for screening HHT drugs. To date our experiments have been performed using only one of each wt, Eng+/- and Eng-/- ES cell lines. To validate our model, multiple new mES cell lines lacking or heterozygous for endoglin will be generated from blastocysts isolated from Eng+/- crosses and EB differentiation in presence of thalidomide will be repeated.
  3. Since we have expertise in the differentiation of multiple human ESC (HESC) lines into mesoderm derivatives, including vascular cells, in EBs we will test the effects of thalidomide in an analogous model to validate and extend our findings on vessel stabilization to human. Time permitting and the context of other studies to optimize siRNA-mediated knockdown in HESCs, we will attempt to generate a human model of HHT development in vitro.

 

The complete results of the Dr. Mummery’s research can be read in Nature Medicine.