The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood-brain barrier.
The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood-brain barrier.
复制标题
DOI：
10.1016/j.nbd.2020.105131
复制
发表时间：
2020-12
影响因子：
6.1
通讯作者：
Ramirez SH
中科院分区：
文献类型：
--
作者：
Buzhdygan TP;DeOre BJ;Baldwin-Leclair A;Bullock TA;McGary HM;Khan JA;Razmpour R;Hale JF;Galie PA;Potula R;Andrews AM;Ramirez SH
随着全球研究人员将注意力集中在了解SARS-CoV-2上，正在出现的画面是一种对包括脑血管系统在内的多个器官系统的血管系统产生严重影响的病毒。观察到的对中枢神经系统的影响包括神经症状（头痛、恶心、头晕）、致命的微凝块形成，在极少数情况下还会引起脑炎。然而，我们对病毒如何引起这些轻微到严重的神经系统症状以及如何影响脑血管系统的理解仍然不清楚。因此，本报告的结果探讨了SARS-CoV-2病毒刺突蛋白对原代人脑微血管内皮细胞（hBMVECs）的有害结果是否可以观察到。刺突蛋白是由含有受体结合域（RBD）的S1亚基和S2亚基组成的，在受体识别中起着关键作用。首先，利用死后脑组织，我们发现血管紧张素转换酶2或ACE2（一种已知的SARS-CoV-2刺突蛋白的结合靶标）在额叶皮层的各种血管口径中普遍表达。此外，在高血压和痴呆患者中，ACE2表达上调。在细胞培养条件下维持的原代hbmvec中也检测到ACE2。细胞活力分析显示，在48小时的暴露窗口内，S1、S2或仅含有RBD的S1的截短形式对hBMVEC活力的影响都很小。将刺突蛋白引入体外血脑屏障模型（BBB）后，其屏障特性发生了显著变化。我们发现的关键是S1促进了人类血脑屏障的高级3D微流控模型的屏障完整性丧失，这是一个更接近于该中枢神经系统界面生理条件的平台。已有证据表明，SARS-CoV-2刺突蛋白会触发脑内皮细胞的促炎反应，这可能导致血脑屏障功能状态的改变。总之，这些结果首次显示了SARS-CoV-2刺突蛋白可能对脑内皮细胞产生的直接影响；从而为COVID-19患者的神经系统后果提供了一个合理的解释。
As researchers across the globe have focused their attention on understanding SARS-CoV-2, the picture that is emerging is that of a virus that has serious effects on the vasculature in multiple organ systems including the cerebral vasculature. Observed effects on the central nervous system include neurological symptoms (headache, nausea, dizziness), fatal microclot formation and in rare cases encephalitis. However, our understanding of how the virus causes these mild to severe neurological symptoms and how the cerebral vasculature is impacted remains unclear. Thus, the results presented in this report explored whether deleterious outcomes from the SARS-CoV-2 viral spike protein on primary human brain microvascular endothelial cells (hBMVECs) could be observed. The spike protein, which plays a key role in receptor recognition, is formed by the S1 subunit containing a receptor binding domain (RBD) and the S2 subunit. First, using postmortem brain tissue, we show that the angiotensin converting enzyme 2 or ACE2 (a known binding target for the SARS-CoV-2 spike protein), is ubiquitously expressed throughout various vessel calibers in the frontal cortex. Moreover, ACE2 expression was upregulated in cases of hypertension and dementia. ACE2 was also detectable in primary hBMVECs maintained under cell culture conditions. Analysis of cell viability revealed that neither the S1, S2 or a truncated form of the S1 containing only the RBD had minimal effects on hBMVEC viability within a 48 h exposure window. Introduction of spike proteins to invitro models of the blood-brain barrier (BBB) showed significant changes to barrier properties. Key to our findings is the demonstration that S1 promotes loss of barrier integrity in an advanced 3D microfluidic model of the human BBB, a platform that more closely resembles the physiological conditions at this CNS interface. Evidence provided suggests that the SARS-CoV-2 spike proteins trigger a pro-inflammatory response on brain endothelial cells that may contribute to an altered state of BBB function. Together, these results are the first to show the direct impact that the SARS-CoV-2 spike protein could have on brain endothelial cells; thereby offering a plausible explanation for the neurological consequences seen in COVID-19 patients.
登录
查看更多内容
DOI：
10.1186/s13195-016-0217-7
发表时间：
2016-11-25
期刊：
Alzheimer's research & therapy
影响因子：
--
作者：
Kehoe PG;Wong S;Al Mulhim N;Palmer LE;Miners JS
通讯作者：
Miners JS
影响因子：
5.4
作者：
Chai, Qingqing;He, Wen Q.;Fu, Zhen F.
通讯作者：
Fu, Zhen F.
影响因子：
28.3
作者：
Gorbalenya, Alexander E.;Baker, Susan C.;Ziebuhr, John
通讯作者：
Ziebuhr, John
影响因子：
168.9
作者：
Chan, Jasper Fuk-Woo;Yuan, Shuofeng;Yuen, Kwok-Yung
通讯作者：
Yuen, Kwok-Yung
影响因子：
6.3
作者：
Andrews, Allison M.;Lutton, Evan M.;Ramirez, Servio H.
通讯作者：
Ramirez, Servio H.