Transcriptome changes during Endothelial to Mesenchymal Transition (EndMT) using a TGF-b2 and IL-1b in vitro model of EndMT in HUVEC and HPAEC. Transcriptome changes during Endothelial to Mesenchymal Transition (EndMT) using a TGF-b2 and IL-1b in vitro model of EndMT in HUVEC and HPAEC

Endothelial-to-mesenchymal transition (EndMT) is a dynamic transformation process that has a functional impact upon pathological vascular remodelling. However, the molecular mechanisms that govern EndMT remain largely unknown. We modelled this process in vitro by exposing human primary endothelial cells to a combination of transforming growth factor-β2 (TGF- β2) and interleukin-1β (IL-1β). RNAseq was carried out to analyse the change of gene expression during the transition and define the transcriptional architecture of EndMT. Overall design: HUVEC (Human umbilical vein endothelial cells) cells were treated with TGF-β2 (10 ng/ml) alone, IL-1β (1ng/ml) alone as well as combined TGF-β2 and IL-1β for 7 consecutive days. HPAEC (Human pulmonary artery endothelial cells) cells were co-treated with TGF-β2 and IL-1β for 7 consecutive days. Control samples correspond to time-matched untreated cells. All treatments were carried out in triplicates and total RNAs were extracted for RNAseq. Ribosomal-depleted stranded libraries were prepared by Beckman Coulter Genomics using the TruSeq Stranded Total RNA kit with Ribo-Zero Gold. Libraries were sequenced with Illumina at an average of 50 million reads per samples (paired end 2 x 125bp).