RESEARCH ARTICLE

CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells

  • Ping Wang 1,4 ,
  • Zunpeng Liu 2,4 ,
  • Xiaoqian Zhang 2,4 ,
  • Jingyi Li 4,5 ,
  • Liang Sun 7 ,
  • Zhenyu Ju 8 ,
  • Jian Li 7 ,
  • Piu Chan 5 ,
  • Guang-Hui Liu , 1,4,5,6,8 ,
  • Weiqi Zhang , 1,4,5 ,
  • Moshi Song , 3,4,6 ,
  • Jing Qu , 2,4,6
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  • 1. National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
  • 2. State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
  • 3. State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
  • 4. University of Chinese Academy of Sciences, Beijing 100049, China
  • 5. National Clinical Research Center for Geriatric Disorders, Xuanwu Hospital of Capital Medical University, Beijing 100053, China
  • 6. Institute of Stem cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China
  • 7. The MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of Gerontology, Beijing 100730, China
  • 8. Key Laboratory of Regenerative Medicine of Ministry of Education, Institute of Aging and Regenerative Medicine, Jinan University, Guangzhou 510632, China

Received date: 28 May 2018

Accepted date: 08 Jun 2018

Published date: 29 Nov 2018

Copyright

2018 The Author(s)

Abstract

Vascular cell functionality is critical to blood vessel homeostasis. Constitutive NF-κB activation in vascular cells results in chronic vascular inflammation, leading to various cardiovascular diseases. However, how NF-κB regulates human blood vessel homeostasis remains largely elusive. Here, using CRISPR/Cas9-mediated gene editing, we generated RelA knockout human embryonic stem cells (hESCs) and differentiated them into various vascular cell derivatives to study how NF-κB modulates human vascular cells under basal and inflammatory conditions. Multi-dimensional phenotypic assessments and transcriptomic analyses revealed that RelA deficiency affected vascular cells via modulating inflammation, survival, vasculogenesis, cell differentiation and extracellular matrix organization in a cell typespecific manner under basal condition, and that RelA protected vascular cells against apoptosis and modulated vascular inflammatory response upon tumor necrosis factor α (TNFα) stimulation. Lastly, further evaluation of gene expression patterns in IκBα knockout vascular cells demonstrated that IκBα acted largely independent of RelA signaling. Taken together, our data reveal a protective role of NF-κB/RelA in modulating human blood vessel homeostasis and map the human vascular transcriptomic landscapes for the discovery of novel therapeutic targets.

Cite this article

Ping Wang , Zunpeng Liu , Xiaoqian Zhang , Jingyi Li , Liang Sun , Zhenyu Ju , Jian Li , Piu Chan , Guang-Hui Liu , Weiqi Zhang , Moshi Song , Jing Qu . CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells[J]. Protein & Cell, 2018 , 9(11) : 945 -965 . DOI: 10.1007/s13238-018-0560-5

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