CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells
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. Her...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Oxford University Press
2018-07-01
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| Series: | Protein & Cell |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1007/s13238-018-0560-5 |
| _version_ | 1797722725953830912 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Ping Wang |
| collection | DOAJ |
| description | 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 type-specific 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. |
| first_indexed | 2024-03-12T09:51:41Z |
| format | Article |
| id | doaj.art-d5f1ef2bc5ec4873bc5d4cb9a2063ba2 |
| institution | Directory Open Access Journal |
| issn | 1674-800X 1674-8018 |
| language | English |
| last_indexed | 2024-03-12T09:51:41Z |
| publishDate | 2018-07-01 |
| publisher | Oxford University Press |
| record_format | Article |
| series | Protein & Cell |
| spelling | doaj.art-d5f1ef2bc5ec4873bc5d4cb9a2063ba22023-09-02T12:29:32ZengOxford University PressProtein & Cell1674-800X1674-80182018-07-0191194596510.1007/s13238-018-0560-5CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cellsPing Wang0Zunpeng Liu1Xiaoqian Zhang2Jingyi Li3Liang Sun4Zhenyu Ju5Jian Li6Piu Chan7Guang-Hui Liu8Weiqi Zhang9Moshi Song10Jing Qu11National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of SciencesState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of SciencesNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesThe MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of GerontologyKey Laboratory of Regenerative Medicine of Ministry of Education, Institute of Aging and Regenerative Medicine, Jinan UniversityThe MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of GerontologyNational Clinical Research Center for Geriatric Disorders, Xuanwu Hospital of Capital Medical UniversityNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesState Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of SciencesState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of SciencesAbstract 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 type-specific 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.http://link.springer.com/article/10.1007/s13238-018-0560-5NF-κBRelAStem cellInflammationApoptosis |
| spellingShingle | 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 Protein & Cell NF-κB RelA Stem cell Inflammation Apoptosis |
| title | CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells |
| title_full | CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells |
| title_fullStr | CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells |
| title_full_unstemmed | CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells |
| title_short | CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells |
| title_sort | crispr cas9 mediated gene knockout reveals a guardian role of nf κb rela in maintaining the homeostasis of human vascular cells |
| topic | NF-κB RelA Stem cell Inflammation Apoptosis |
| url | http://link.springer.com/article/10.1007/s13238-018-0560-5 |
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