PMID- 30354255 OWN - NLM STAT- MEDLINE DCOM- 20190719 LR - 20220129 IS - 1524-4636 (Electronic) IS - 1079-5642 (Linking) VI - 38 IP - 9 DP - 2018 Sep TI - BDNF (Brain-Derived Neurotrophic Factor) Promotes Embryonic Stem Cells Differentiation to Endothelial Cells Via a Molecular Pathway, Including MicroRNA-214, EZH2 (Enhancer of Zeste Homolog 2), and eNOS (Endothelial Nitric Oxide Synthase). PG - 2117-2125 LID - 10.1161/ATVBAHA.118.311400 [doi] AB - Objective- The NTs (neurotrophins), BDNF (brain-derived neurotrophic factor) and NT-3 promote vascular development and angiogenesis. This study investigated the contribution of endogenous NTs in embryonic stem cell (ESC) vascular differentiation and the potential of exogenous BDNF to improve the process of ESC differentiation to endothelial cells (ECs). Approach and Results- Mouse ESCs were differentiated into vascular cells using a 2-dimensional embryoid body (EB) model. Supplementation of either BDNF or NT-3 increased EC progenitors' abundance at day 7 and enlarged the peripheral vascular plexus with ECs and SM22alpha(+) (smooth muscle 22 alpha-positive) smooth muscle cells by day 13. Conversely, inhibition of either BDNF or NT-3 receptor signaling reduced ECs, without affecting smooth muscle cells spread. This suggests that during vascular development, endogenous NTs are especially relevant for endothelial differentiation. At mechanistic level, we have identified that BDNF-driven ESC-endothelial differentiation is mediated by a pathway encompassing the transcriptional repressor EZH2 (enhancer of zeste homolog 2), microRNA-214 (miR-214), and eNOS (endothelial nitric oxide synthase). It was known that eNOS, which is needed for endothelial differentiation, can be transcriptionally repressed by EZH2. In turn, miR-214 targets EZH2 for inhibition. We newly found that in ESC-ECs, BDNF increases miR-214 expression, reduces EZH2 occupancy of the eNOS promoter, and increases eNOS expression. Moreover, we found that NRP-1 (neuropilin 1), KDR (kinase insert domain receptor), and pCas(130) (p130 Crk-associated substrate kinase), which reportedly induce definitive endothelial differentiation of pluripotent cells, were increased in BDNF-conditioned ESC-EC. Mechanistically, miR-214 mediated the BDNF-induced expressional changes, contributing to BDNF-driven endothelial differentiation. Finally, BDNF-conditioned ESC-ECs promoted angiogenesis in vitro and in vivo. Conclusions- BDNF promotes ESC-endothelial differentiation acting via miR-214. FAU - Descamps, Betty AU - Descamps B AD - From the Bristol Heart Institute, School of Clinical Sciences, University of Bristol, United Kingdom (B.D., J.S., G.B., C.E.). FAU - Saif, Jaimy AU - Saif J AD - From the Bristol Heart Institute, School of Clinical Sciences, University of Bristol, United Kingdom (B.D., J.S., G.B., C.E.). FAU - Benest, Andrew V AU - Benest AV AD - Tumour and Vascular Biology Laboratories, Cancer Biology, Division of Cancer and Stem Cells, School of Medicine, University of Nottingham, United Kingdom (A.V.B., D.O.B.). FAU - Biglino, Giovanni AU - Biglino G AD - From the Bristol Heart Institute, School of Clinical Sciences, University of Bristol, United Kingdom (B.D., J.S., G.B., C.E.). FAU - Bates, David O AU - Bates DO AD - Tumour and Vascular Biology Laboratories, Cancer Biology, Division of Cancer and Stem Cells, School of Medicine, University of Nottingham, United Kingdom (A.V.B., D.O.B.). FAU - Chamorro-Jorganes, Aranzazu AU - Chamorro-Jorganes A AD - National Heart and Lung Institute, Imperial College London, United Kingdom (A.C.-J., C.E.). FAU - Emanueli, Costanza AU - Emanueli C AD - From the Bristol Heart Institute, School of Clinical Sciences, University of Bristol, United Kingdom (B.D., J.S., G.B., C.E.). AD - National Heart and Lung Institute, Imperial College London, United Kingdom (A.C.-J., C.E.). LA - eng GR - MR/K013157/1/MRC_/Medical Research Council/United Kingdom GR - CH/15/1/31199/BHF_/British Heart Foundation/United Kingdom GR - PG/11/67/29067/BHF_/British Heart Foundation/United Kingdom PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - Arterioscler Thromb Vasc Biol JT - Arteriosclerosis, thrombosis, and vascular biology JID - 9505803 RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Crk-Associated Substrate Protein) RN - 0 (MicroRNAs) RN - 0 (Mirn214 microRNA, mouse) RN - 0 (Nerve Growth Factors) RN - 0 (neurotropin 3, mouse) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type III) RN - EC 2.1.1.43 (Enhancer of Zeste Homolog 2 Protein) RN - EC 2.1.1.43 (Ezh2 protein, mouse) RN - EC 2.7.10.1 (Kdr protein, mouse) RN - EC 2.7.10.1 (Vascular Endothelial Growth Factor Receptor-2) RN - EC 5.2.1.8 (Immunophilins) SB - IM MH - Animals MH - Brain-Derived Neurotrophic Factor/pharmacology/*physiology MH - *Cell Differentiation/drug effects MH - Cell Line MH - Crk-Associated Substrate Protein/metabolism MH - Embryonic Stem Cells/*physiology MH - Endothelial Cells/*physiology MH - Enhancer of Zeste Homolog 2 Protein/antagonists & inhibitors/*metabolism MH - Immunophilins/metabolism MH - Mice MH - MicroRNAs/*metabolism MH - Muscle, Smooth, Vascular/cytology/metabolism MH - Myocytes, Smooth Muscle/metabolism MH - *Neovascularization, Physiologic MH - Nerve Growth Factors/pharmacology MH - Nitric Oxide Synthase Type III/*metabolism MH - Vascular Endothelial Growth Factor Receptor-2/metabolism OTO - NOTNLM OT - angiogenesis OT - brain-derived neurotrophic factor OT - embryonic stem cells OT - endothelial differentiation OT - microRNAs EDAT- 2018/10/26 06:00 MHDA- 2019/07/20 06:00 CRDT- 2018/10/26 06:00 PHST- 2018/10/26 06:00 [entrez] PHST- 2018/10/26 06:00 [pubmed] PHST- 2019/07/20 06:00 [medline] AID - 10.1161/ATVBAHA.118.311400 [doi] PST - ppublish SO - Arterioscler Thromb Vasc Biol. 2018 Sep;38(9):2117-2125. doi: 10.1161/ATVBAHA.118.311400.