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6月，中國科學院生物物理研究所中國科學院生物大分子**中心生物大分子國家實驗室；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；中國科學院動物研究所膜生物學國家重點實驗室；中國科學院大學；首都醫科大學宣武醫院國家老年病臨床研究中心；中國科學院干細胞與再生研究所；國家老年醫學中心北京醫院衛生部老年醫學重點實驗室；暨南大學衰老與再生醫學研究所再生醫學教育部重點實驗室（National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China；State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China；State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China；University of Chinese Academy of Sciences, Beijing 100049,   China；National Clinical Research Center for Geriatric Disorders, Xuanwu Hospital of Capital Medical University,   Beijing 100053,China；Institute of Stem cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China；The MOH Key Laboratory of Geriatrics, Beijing Hospital,   National Center of Gerontology, Beijing 100730, China；Key Laboratory of Regenerative Medicine of Ministry of Education, Institute of Aging and Regenerative Medicine, Jinan University, Guangzhou 510632, China) Guang-Hui Liu老師研究團隊在《Protein& Cell》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：

內聚力1同源物2 (ESCO2)是調控內聚力和基因組穩定性的重要基因，目前尚未在人胃癌(GC)中進行研究。科研人員發現ESCO2在體外顯著抑制人胃癌細胞系細胞增殖和誘導細胞凋亡，并在體內抑制腫瘤異種移植物的發育。此外，腺苷單磷酸活化蛋白激酶(AMPK)在抑制其下游靶點后被激活，包括哺乳動物雷帕霉素靶點(mTOR)和p70核糖體S6激酶1 (p70S6K1)，這一結果與p53的激活一致。值得注意的是，共免疫沉淀(Co-IP)分析表明，ESCO2可以與GC細胞中的p53相互作用。綜上所述，科研人員的數據表明，ESCO2對胃癌的發展至關重要，可能是治療胃癌的潛在治療靶點。

該論文中，人永生化胃上皮細胞系GES-1和人胃癌細胞系AGS、SGC-7901、MKN-45、BGC-823、MGC-803、HGC-27和MKN-74細胞的體外培養是使用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.