Nitric oxide-mediated changes in glymphatic and CSF systems in aging and Alzheimer's disease

摘要 SARS-CoV-2与机体的接触始于与其关键受体ACE2的结合，ACE2是高度 表达于气管、支气管和鼻腔粘液纤毛上皮的纤毛细胞。绑定后， 该病毒抑制了呼吸道中的纤毛跳动，并抑制了粘液纤毛的运输，第一行 防御入侵的微生物。因此，增强睫毛功能可能会预防病毒 在最初的一步就感染病毒或帮助阻止病毒的进一步传播。还表达了ACE2 在选定的脑细胞中，研究了SARS-CoV-2对神经系统的影响。 这一补充的主要目标是快速建立一个筛选系统，以识别能够 立即申请新冠肺炎治疗，并用所学知识拓展 筛选范围更广的候选化合物。 在我们的NIA资助的赠款中，我们发现了nNOS-NO-CGC-cGMP信号的关键作用 大脑和呼吸道中纤毛活动的通路。在这里，我们描述了我们针对这一目标的计划 使用临床批准的药物，以增强纤毛活动和呼吸道清除。这 该项目可以稍后扩展，以构建灵敏而强大的抗新冠肺炎治疗药物筛选。 在我们的第一个具体目标中，我们将开发一个全面的计算工具箱，并使用它来 确定呼吸道和脑纤毛细胞纤毛活动的关键参数(标志) 从人类和人性化的小鼠模型。在第二个具体目标中，我们将把我们的设置应用于 确定对SARS-CoV-2的主要细胞结合配体S1的纤毛反应的签名，并 S1-表达假型VSV-S1病毒。最后，在我们的第三个具体目标中，我们将检查反 目前临床上使用的SARS-CoV-2药物和临床批准的激活NO-2的化合物- CGMP途径，以及它们的组合，以确定这种治疗是否可以拯救受S1影响的 纤毛活动在呼吸道和大脑中的准备。在同一组实验中，我们将检查 来自不同年龄动物的制剂的反应，这可能有助于深刻的老年- 新冠肺炎的偏向。我们强调，我们选择的测试药物仅限于 经治疗批准的化合物，从而使我们的发现能够快速转化为临床 练习一下。

ABSTRACT SARS-CoV-2 contact with the organism starts with binding to its key receptor ACE2 which is highly expressed in the ciliated cells of the tracheal, bronchial, and nasal mucociliary epithelium. After binding, the virus inhibits the cilia beating in the airways and suppresses mucociliary transport, the first line of defense against invading microorganisms. Thus, augmenting the ciliary function may prevent the viral infection at the very first steps or help fight the further propagation of the virus. ACE2 is also expressed in selected brain cells, underlying the neurological effects of SARS-CoV-2. The main objective of this supplement is to rapidly build a screening system, to identify drugs that can be immediately applied for COVID-19 therapy, and to use the gained knowledge for expanding the screen to a broader range of candidate compounds. In our NIA supported grant we discovered the critical role of the nNOS-NO-cGC-cGMP signaling pathway for ciliary activity in the brain and the airways. Here we describe our plans of targeting this pathway with clinically-approved drugs in order to augment cilia activity and airways clearance. This project can be later extended to build a sensitive and powerful drug screen for anti-COVID-19 therapy. In our first Specific Aim we will develop a comprehensive computational toolbox and use it to determine the key parameters (a signature) of the cilia activity in the airways’ and brain ciliated cells from humans and humanized mouse model. In the second Specific Aim, we will apply our setup to determine the signature of the cilia response to S1, the main cell binding ligand of SARS-CoV-2, and to S1-expressing pseudotyped VSV-S1 virus. Finally, in our third Specific Aim we will examine anti- SARS-CoV-2 drugs currently used in clinic and clinically-approved compounds that activate the NO- cGMP pathway, as well as their combination to determine if such treatments can rescue the S1-affected cilia activity in the airways and brain preparations. In the same set of experiments we will examine the response of preparations from animals of different age, which may contribute to the profound old age- bias of COVID-19. We emphasize that our selection of drugs to be tested is limited to the therapeutically-approved compounds, thus enabling rapid translation of our findings into clinical practice.