Chromatin-modifying agents convert fibroblasts to OCT4+ and VEGFR-2+ capillary tube-forming cells

• Published in: PloS one Vol. 12; no. 5; p. e0176496
• Main Authors: Wary, Anita, Wary, Neil, Baruah, Jugajyoti, Mastej, Victoria, Wary, Kishore K
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 03.05.2017; Public Library of Science (PLoS)
• Subjects: 5-aza-2'-deoxycytidine; Biology and Life Sciences; Capillaries; Capillaries - cytology; Capillary tubes; Cell adhesion; Cell adhesion & migration; Cell cycle; Cells, Cultured; Chromatin; Chromatin - metabolism; Chromatin Immunoprecipitation; Endothelial cells; Epigenetics; Fetuses; Fibroblasts; Fibroblasts - metabolism; Forming; Gene expression; Growth; Humans; Immunoprecipitation; Ischemia; Kinases; Liver; Medicine and Health Sciences; Microscopic analysis; Nuclei; Nuclei (cytology); Oct-4 protein; Octamer Transcription Factor-3 - metabolism; Pharmacology; Phosphatase; Phosphorylation; Physiological aspects; Plastics; Pluripotency; Research and Analysis Methods; Signaling; Skin; Stem cells; Substrates; Trichostatin A; Vascular endothelial growth factor; Vascular endothelial growth factor receptor 2; Vascular Endothelial Growth Factor Receptor-2 - metabolism; Vascular endothelial growth factor receptors; Wnt protein; β-catenin; United States > US; Illinois; Chicago Illinois
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0176496

The human epigenome is plastic. The goal of this study was to address if fibroblast cells can be epigenetically modified to promote neovessel formation. Here, we used highly abundant human adult dermal fibroblast cells (hADFCs) that were treated with the chromatin-modifying agents 5-aza-2'-deoxycytidine and trichostatin A, and subsequently subjected to differentiation by activating Wnt signaling. Our results show that these epigenetically modified hADFCs increasingly expressed β-catenin, pluripotency factor octamer-binding transcription factor-4 (OCT4, also known as POU5F1), and endothelial cell (EC) marker called vascular endothelial growth factor receptor-2 (VEGFR-2, also known as Fetal Liver Kinase-1). In microscopic analysis, β-catenin localized to cell-cell contact points, while OCT4 was found to be localized primarily to the nucleus of these cells. Furthermore, in a chromatin immunoprecipitation experiment, OCT4 bound to the VEGFR-2/FLK1 promoter. Finally, these modified hADFCs also transduced Wnt signaling. Importantly, on a two-dimensional (2D) gel substrate, a subset of the converted cells formed vascular network-like structures in the presence of VEGF. Chromatin-modifying agents converted hADFCs to OCT4+ and VEGFR-2+ capillary tube-forming cells in a 2D matrix in VEGF-dependent manner.