CRISPR/ cas9介導(dǎo)的基因敲除揭示了NF-κB/RelA在維持人血管細(xì)胞穩(wěn)態(tài)中的守護(hù)作用

2018年7月，1中國(guó)科學(xué)院生物物理研究所，中國(guó)科學(xué)院生物大分子**研究中心，中國(guó)科學(xué)院生物大分子國(guó)家實(shí)驗(yàn)室，2中國(guó)科學(xué)院動(dòng)物研究所干細(xì)胞與生殖生物學(xué)國(guó)家重點(diǎn)實(shí)驗(yàn)室，3中國(guó)科學(xué)院動(dòng)物研究所膜生物學(xué)國(guó)家重點(diǎn)實(shí)驗(yàn)室，4中國(guó)科學(xué)院大學(xué)，5首都醫(yī)科大學(xué)宣武醫(yī)院國(guó)家老年疾病臨床研究中心，6中國(guó)科學(xué)院干細(xì)胞與再生研究所，7國(guó)家老年醫(yī)學(xué)中心，北京醫(yī)院，衛(wèi)生部老年醫(yī)學(xué)重點(diǎn)實(shí)驗(yàn)室，8暨南大學(xué)再生醫(yī)學(xué)研究所再生醫(yī)學(xué)教育部重點(diǎn)實(shí)驗(yàn)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(tuán)隊(duì)在《Inflammation》上發(fā)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(dǎo)的基因敲除揭示了NF-κB/RelA在維持人血管細(xì)胞穩(wěn)態(tài)中的守護(hù)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(xì)胞的功能對(duì)血管穩(wěn)態(tài)至關(guān)重要。血管細(xì)胞的構(gòu)造性NF-κB活化導(dǎo)致慢性血管炎癥，導(dǎo)致多種心血管疾病。然而，NF-κB如何調(diào)節(jié)人體血管穩(wěn)態(tài)在很大程度上仍然是一個(gè)謎。在這里，我們利用CRISPR/ cas9介導(dǎo)的基因編輯技術(shù)，生成RelA敲除的人胚胎干細(xì)胞(hESCs)，并將其分化為各種血管細(xì)胞衍生物，研究NF-κB在基礎(chǔ)和炎癥條件下如何調(diào)節(jié)人血管細(xì)胞。多維表型分析和轉(zhuǎn)錄組學(xué)分析顯示，RelA缺乏在基礎(chǔ)條件下通過(guò)細(xì)胞特異性方式調(diào)節(jié)血管細(xì)胞的炎癥、存活、血管發(fā)生、細(xì)胞分化和細(xì)胞外基質(zhì)組織，并在腫瘤壞死因子α (TNFα)刺激下保護(hù)血管細(xì)胞免受凋亡和調(diào)節(jié)血管炎癥反應(yīng)。最后，對(duì)i-κbα敲除血管細(xì)胞基因表達(dá)模式的進(jìn)一步評(píng)估表明，i - κ b α在很大程度上獨(dú)立于RelA信號(hào)傳導(dǎo)。綜上所述，我們的數(shù)據(jù)揭示了NF-κB/RelA在調(diào)節(jié)人類血管穩(wěn)態(tài)中的保護(hù)作用，并為發(fā)現(xiàn)新的治療靶點(diǎn)繪制了人類血管轉(zhuǎn)錄組圖譜。

該論文中，H9 human ESCs體外培養(yǎng)是使用Ausbian特級(jí)胎牛血清完成的。欲了解或購(gòu)買(mǎi)Ausbian特級(jí)胎牛血清可以聯(lián)系北京締一生物400-166-8600.

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