Preclinical studies to establish the combination of apoA-I mimetic peptides and statins as novel therapy for COVID-19

项目摘要/摘要 SARS-CoV-2大流行强调了迫切需要确定可以被新的 抗病毒药物。这项建议描述了apoA-I模拟物与他汀类药物相结合作为新疗法的使用 为了新冠肺炎。SARS-CoV-2利用线粒体在肺上皮细胞中复制。线粒体反应性 氧物种(mito-ROS)诱导脂质、膜和氧化还原敏感蛋白的改变，最终 病毒复制。因此，我们假设抗氧化剂ApoA-I模拟多肽4F，它针对脂质， Mito-Ros和氧化还原途径，对SARS-CoV-2具有抗病毒活性。我们确认4F不仅有 对SARS-CoV-2有抗病毒活性，但对上皮细胞也有抗炎作用，这可能会缓解 新冠肺炎中的肺损伤。重要的是，先前的研究表明，4F和他汀类药物可能在 对引发心血管疾病的血脂、氧化应激和炎症的体内效应。 人体研究表明，他汀类药物的使用与患严重新冠肺炎的风险较低有关。两者都有 他汀类药物和4F减弱Toll样受体-CD147-NF-κB通路的激活，这是关键 冠状病毒感染中的炎症途径。我们假设4F和他汀类药物的结合 肺上皮细胞抗SARS-CoV-2活性和抗炎活性 细胞、巨噬细胞和血管内皮细胞通过减弱氧化还原敏感型TLR-CD147-NF-κB 前驱和促炎途径。因此，在本提案的目标1中，使用气-液界面(ALI) 原代呼吸道上皮细胞和细胞系的培养，结合病毒和免疫分析，我们将 确定4F与阿托伐他汀联合用药抑制心肌细胞异常激活的机制 TLR-CD147-NF-κB途径作为驱动病毒复制和相关炎症的新机制 SARS-CoV-2感染肺细胞的应答。鉴于血管功能障碍可能是 新冠肺炎慢性感染后后遗症，在本建议的目标2中，使用已建立的体外试验 动脉粥样硬化和血管功能障碍模型及外周血单核细胞的研究 新冠肺炎患者结合免疫检测，我们将确定4F和阿托伐他汀 抑制新冠肺炎患者巨噬细胞TLR-CD147-NF-κB通路的异常激活 在体外与内皮细胞相互作用，以驱动促炎症、促动脉粥样硬化反应。我们的独立派 AIMS将相辅相成，并将促进apoA-I模拟多肽和 他汀类药物作为新冠肺炎的新疗法。这项工作具有创新性、跨学科、面向公共卫生和 直接解决融资机会的目标。

PROJECT SUMMARY/ABSTRACT The SARS-CoV-2 pandemic emphasizes the urgent need to determine pathways that can be targeted by novel antivirals. This proposal describes the use of the combination of apoA-I mimetics with statins as novel therapy for COVID-19. SARS-CoV-2 utilizes mitochondria to replicate in lung epithelial cells. Mitochondrial reactive oxygen species (mito-ROS) induce alterations of lipids, membranes and redox sensitive proteins and ultimately viral replication. Thus, we hypothesized that the antioxidant ApoA-I mimetic peptide 4F, that targets lipids, mito-ROS and redox pathways, has antiviral activity against SARS-CoV-2. We confirmed that 4F not only has antiviral activity against SARS-CoV-2 but also has anti-inflammatory effects in epithelial cells that may alleviate lung injury in COVID-19. Importantly, prior studies have shown that 4F and statins may have major additive in vivo effects on altered lipids, oxidative stress and inflammation that are instigators of cardiovascular disease. Human studies suggest that statin use was associated with a lower risk of developing severe COVID-19. Both statins and 4F attenuate activation of the Toll like Receptor (TLR)-CD147-NF-κB pathways, which are key inflammatory pathways in coronavirus infections. We hypothesize that the combination of 4F and statins has antiviral activity against SARS-CoV-2 in lung epithelial cells and anti-inflammatory activity in lung epithelial cells, macrophages and vascular endothelial cells by attenuating the redox sensitive TLR-CD147-NF-κB proviral and proinflammatory pathway. Thus, in Aim 1 of this proposal and using an air-liquid interface (ALI) culture of primary airway epithelial cells, and cell lines in combination with viral and immune assays, we will determine the mechanisms how the combination of 4F with atorvastatin attenuates aberrant activation of the TLR-CD147-NF-κB pathway as a novel mechanism that drives viral replication and associated inflammatory responses in SARS-CoV-2 infected lung cells. Given that vascular dysfunction is a possible mechanism of chronic post-infectious sequalae of COVID-19, in Aim 2 of this proposal and using an established ex vivo model of atherogenesis and vascular dysfunction and peripheral blood mononuclear cells (PBMCs) from COVID-19 patients in combination with immune assays, we will determine whether 4F and atorvastatin attenuate aberrant activation of the TLR-CD147-NF-κB pathway in macrophages from COVID-19 patients that interact ex vivo with endothelial cells to drive proinflammatory proatherogenic responses. Our independent aims will complement each other and will advance the use of the combination of apoA-I mimetic peptides and statins as novel therapy for COVID-19. This work is innovative, interdisciplinary, public health-oriented, and directly addresses the goals of funding opportunity.