Multiplexed imaging of Renin-Angiotensin System (RAS) pathways, endothelial and immune dysfunction in COVID-19 Lung

项目总结 SARS-CoV2大流行已经造成了很高的伤亡，约70万人(约15.5万美国人)死亡，18人死亡 全球有100万(约500万美国)感染者，在许多国家和新的热点地区蔓延有增无减 定期重新出现。虽然人们正在疯狂地努力开发疫苗和治疗方法 人们很有希望在未来6-12个月内提供这些产品，目前还没有答案 这些措施将在多大程度上有效，多长时间有效。对这种疾病的持续了解 病原学，特别是首先感染的器官，并因此确定新的干预途径 是很重要的。病毒感染的主要途径是通过呼吸道，据报道病毒是 通过血管渗漏直接(通过感染)或间接(通过)通过肺传播到其他器官 损害血管紧张素转换酶2活性)影响内皮细胞和免疫细胞。表达ACE2酶的细胞 和其他病毒辅助受体(TMPRSS2或组织蛋白酶L)是病毒感染的主要目标。ACE2 在肾素-血管紧张素系统(RAS)途径的调节中起关键作用。通过将 血管紧张素转换酶活性的乘积，血管紧张素II(Ang II)与血管紧张素1-7(Ang 1-7)的乘积，ACE2减少Ang II 介导的有害作用，包括促进血管壁炎症、内皮 功能障碍，内皮细胞和血管平滑肌细胞迁移，生长，增殖和 血栓形成。病毒结合破坏ACE2可能会降低这种保护作用。对人类的抑制 一氧化氮的产生、巨核细胞、补体和血小板的激活也可导致 血栓形成和溶栓功能障碍导致肺动脉和其他器官形成血栓。 自从早期史无前例的全球努力识别SARS-CoV2的细胞靶点以来 来自多个人类和非人类单细胞数据集的细胞RNA测序(ScRNAseq)数据， 关于单个器官感染以及受此影响的细胞和细胞途径的几个深入报道 病毒已经出现了。这些报告中的大多数是基于同源或同源基因的转录分析。 虽然报告了一些单链免疫荧光分析，但仍有分类样本。 新冠肺炎组织学显示一种非常异质性的疾病，这可能是一种 这些因素包括细胞组成、空间组织和邻近细胞的激活。至 了解这些因素，我们在这里建议对SARS进行原位多重免疫荧光研究- CoV2阳性和阴性患者样本的空间分布 下呼吸道与血管内皮细胞和免疫细胞中RAS通路激活的作用 功能障碍与新冠肺炎的发病率和死亡率有关。

PROJECT SUMMARY SARS-CoV2 pandemic has already taken a high toll with ~700,000 (~155,00 US) deaths and 18 million (~5 million US) infections globally with unabated spread in many countries and new hotspots reemerging on a regular basis. While frantic efforts are underway to develop vaccines and therapies and there is high hope that these will be available over the next 6-12 months, there are no answers to how much and how long these will be effective. A continued effort on understanding the disease etiology, particularly in the organ first infected, and identifying new avenues of intervention therefore is important. The major route of virus infection is via the respiratory tract and virus is reported to spread via lung to other organs by vascular leakage by directly (through infection) or indirectly (by impairing ACE2 activity) affecting the endothelial and immune cells. Cells expressing ACE2 enzyme and other viral coreceptors (TMPRSS2 or Cathepsin L) are the major targets of viral infection. ACE2 is a key player in regulation of the Renin-Angiotensin system (RAS) pathway. By converting the product of ACE activity, angiotensin II (ang II), to angiotensin 1-7 (ang 1-7), ACE2 diminishes Ang II mediated deleterious effects that can include promoting vascular wall inflammation, endothelial dysfunction, endothelial cell and vascular smooth muscle cell migration, growth, proliferation, and thrombosis. Disruption of ACE2 by viral binding may reduce this protective effect. The inhibition of nitric oxide production, activation of megakaryocytes, complement and platelets can also cause thrombosis and thrombolytic dysfunction leading to clot formation in lung arteries and other organs. Since the early unprecedented global effort to identify the cellular targets of SARS-CoV2 using single cell RNA sequencing (scRNAseq) data from multiple human and non-human single cell datasets, several in depth reports on individual organs infected and cells and cellular pathways affected by this virus have appeared. Most of these reports are based on transcriptomic analysis of homogenized or disaggregated samples although some singleplex immunofluorescence analysis have been reported. COVID-19 tissue histology shows a very heterogenous disease which may be a function of multiple factors including cellular composition, spatial organization and neighboring cell activation. To understand these factors, here we propose an in situ multiplex immunofluorescence study of SARS- CoV2 positive and negative patient samples to spatially profile the cell types affected in the upper and lower respiratory tract and the role of RAS pathway activation in endothelial and immune cell dysfunction related to COVID-19 morbidity and mortality.