cAMP/EPAC Signaling Enables ETV2 to Induce Endothelial Cells with High Angiogenesis Potential

Although the generation of ETV2-induced endothelial cells (iECs) from human fibroblasts serves as a novel therapeutic strategy in regenerative medicine, the process is inefficient, resulting in incomplete iEC angiogenesis. Therefore, we employed chromatin immunoprecipitation (ChIP) sequencing and id...

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Bibliographic Details
Published in:Molecular therapy 2020-02, Vol.28 (2), p.466-478
Main Authors: Kim, Jae-Jun, Kim, Da-Hyun, Lee, Jin Young, Lee, Byung-Chul, Kang, Insung, Kook, Myung Geun, Kong, Dasom, Choi, Soon Won, Woo, Heung-Myong, Kim, Dong-Ik, Kang, Kyung-Sun
Format: Article
Language:English
Subjects:
angiogenesis
cAMP/EPAC/RAP1 pathway
Cellular Reprogramming - genetics
Cyclic AMP - metabolism
direct reprogramming
Ectopic Gene Expression
endothelial cells
Endothelial Cells - metabolism
ETV2
Fibroblasts - metabolism
Guanine Nucleotide Exchange Factors - metabolism
Human Umbilical Vein Endothelial Cells
Humans
Immunohistochemistry
Ischemia - genetics
Ischemia - metabolism
Ischemia - pathology
Neovascularization, Physiologic
Original
rap1 GTP-Binding Proteins - metabolism
Signal Transduction
Transcription Factors - genetics
Transcription Factors - metabolism
transdifferentiation
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Summary:Although the generation of ETV2-induced endothelial cells (iECs) from human fibroblasts serves as a novel therapeutic strategy in regenerative medicine, the process is inefficient, resulting in incomplete iEC angiogenesis. Therefore, we employed chromatin immunoprecipitation (ChIP) sequencing and identified molecular mechanisms underlying ETV2-mediated endothelial transdifferentiation to efficiently produce iECs retaining appropriate functionality in long-term culture. We revealed that the majority of ETV2 targets in human fibroblasts are related to vasculature development and signaling transduction pathways, including Rap1 signaling. From a screening of signaling pathway modulators, we confirmed that forskolin facilitated efficient and rapid iEC reprogramming via activation of the cyclic AMP (cAMP)/exchange proteins directly activated by cAMP (EPAC)/RAP1 axis. The iECs obtained via cAMP signaling activation showed superior angiogenesis in vivo as well as in vitro. Moreover, these cells could form aligned endothelium along the vascular lumen ex vivo when seeded into decellularized liver scaffold. Overall, our study provided evidence that the cAMP/EPAC/RAP1 axis is required for the efficient generation of iECs with angiogenesis potential. [Display omitted] Kim et al. identify that ETV2 directly binds to RAPGEF3, encoding EPAC1, during endothelial transdifferentiation by using genome-wide ChIP-seq. Given these results, they employ forskolin, a cAMP activator, to increase the efficiency of endothelial reprogramming through activation of the cAMP/EPAC1/RAP1 pathway. They also report that forskolin-treated iECs acquire functional features of vascular ECs following long-term culture.
ISSN:1525-0016
1525-0024
DOI:10.1016/j.ymthe.2019.11.019