CRISPR/ cas9介導的基因敲除揭示了NF-κB/RelA在維持人血管細胞穩態中的守護作用

2018年7月，1中國科學院生物物理研究所，中國科學院生物大分子**研究中心，中國科學院生物大分子國家實驗室，2中國科學院動物研究所干細胞與生殖生物學國家重點實驗室，3中國科學院動物研究所膜生物學國家重點實驗室，4中國科學院大學，5首都醫科大學宣武醫院國家老年疾病臨床研究中心，6中國科學院干細胞與再生研究所，7國家老年醫學中心，北京醫院，衛生部老年醫學重點實驗室，8暨南大學再生醫學研究所再生醫學教育部重點實驗室，（1National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics,Chinese Academy of Sciences,

2 State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences,

3 State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences,

4 University of Chinese Academy of Sciences,

5 National Clinical Research Center for Geriatric Disorders, Xuanwu Hospital of Capital Medical University,

6 Institute of Stem cell and Regeneration, Chinese Academy of Sciences,

7 The MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of Gerontology,

8 Key Laboratory of Regenerative Medicine of Ministry of Education, Institute of ging and Regenerative Medicine, JinanUniversity,）

Jing Qu研究團隊在《Inflammation》上發表論文：“CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells”

“CRISPR/ cas9介導的基因敲除揭示了NF-κB/RelA在維持人血管細胞穩態中的守護作用”

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.

摘要

血管細胞的功能對血管穩態至關重要。血管細胞的構造性NF-κB活化導致慢性血管炎癥，導致多種心血管疾病。然而，NF-κB如何調節人體血管穩態在很大程度上仍然是一個謎。在這里，我們利用CRISPR/ cas9介導的基因編輯技術，生成RelA敲除的人胚胎干細胞(hESCs)，并將其分化為各種血管細胞衍生物，研究NF-κB在基礎和炎癥條件下如何調節人血管細胞。多維表型分析和轉錄組學分析顯示，RelA缺乏在基礎條件下通過細胞特異性方式調節血管細胞的炎癥、存活、血管發生、細胞分化和細胞外基質組織，并在腫瘤壞死因子α (TNFα)刺激下保護血管細胞免受凋亡和調節血管炎癥反應。最后，對i-κbα敲除血管細胞基因表達模式的進一步評估表明，i - κ b α在很大程度上獨立于RelA信號傳導。綜上所述，我們的數據揭示了NF-κB/RelA在調節人類血管穩態中的保護作用，并為發現新的治療靶點繪制了人類血管轉錄組圖譜。

該論文中，H9 human ESCs體外培養是使用Ausbian特級胎牛血清完成的。欲了解或購買Ausbian特級胎牛血清可以聯系北京締一生物400-166-8600.

|