Develop broad-spectrum antiviral agents against COVID-19 based on innate immune response to SARS-CoV-2 infection

摘要 这项建议的目标是确定I型干扰素(干扰素-I)介导的抗病毒基因程序 抗SARS-CoV-2感染和开发新型广谱抗病毒药物(BSAAs) 新冠肺炎和其他新发传染病的风险。目前还没有有效的治疗药物。 可用于抗击全球新冠肺炎大流行，在这次疫情中，SARS-CoV-2已感染数百万人 确诊病例增加，并造成数十万人死亡。针对单一病毒的药物，如 HIV逆转录酶和流感神经氨酸酶的抑制剂，需要全面的 对病毒的生命周期和致病机制的了解，使这些药物的开发 这必然很耗时。由新出现的高致病性病毒引起的感染暴发， 包括禽流感、SARS、埃博拉、寨卡病毒(ZIKV)和SARS-CoV-2，已成为#年的主要关切 在过去的二十年里。我们不能依靠传统的针对病毒的药物来治疗由这些疾病引起的疾病 不可预测的新出现的病毒。因此，制定有效的BSA来预防 一系列病毒。我的实验室一直在研究抗病毒的先天免疫反应，特别是干扰素-1 信号通路及其下游基因计划，在过去的20年。虽然该领域之前已经 只关注干扰素刺激的基因(ISGs)，我们已经证明ISGs类似于胆固醇25- 羟基酶(CH25H)和干扰素-I下调的基因，如脂肪酸合成酶(FASN)都起着重要作用 在限制病毒感染和复制方面。我们还确定了多个小分子作为BSA使用， 包括CH25H的代谢产物25-羟基胆甾醇(25HC)和FASN抑制剂C75。在……里面 此外，我们广泛研究了宿主对冠状病毒感染的先天免疫反应：我们有 发表了多篇论文，解释了宿主干扰素-I信号转导通路是如何被激活的 对冠状病毒如小鼠肝炎病毒(MHV)的感染以及冠状病毒如何抑制其 宿主的先天免疫反应是通过病毒木瓜蛋白样蛋白酶(PLPro)实现的。最重要的是，在我们的 初步研究发现，25HC和C75对SARS-CoV-2均有较强的抑制作用。 感染。我们推测，干扰素-I介导的抗病毒基因程序不仅涉及上调 抗病毒ISGs还下调了病毒感染和复制所需的宿主基因。我们进一步 假设通过识别干扰素-I介导的抗SARS-CoV-2基因程序，我们将能够 开发新型抗病毒药物，以对抗新冠肺炎和其他新兴威胁。在这项提案中，我们将首先 确定SARS-CoV-2天然免疫应答中的干扰素-I基因程序。我们还将开发25HC， 25HC类似物和FASN抑制剂作为新型抗SARS-CoV-2药物。我们相信我们的研究将会 不仅确定干扰素-I介导的抗病毒基因在宿主对SARS-CoV-2的天然免疫应答中的作用 不仅如此，还可以发展BSA来治疗新冠肺炎和其他新出现的传染病。

ABSTRACT The goals of this proposal are to determine the type I interferon (IFN-I)-mediated antiviral gene program against SARS-CoV-2 infection and to develop novel broad-spectrum antiviral agents (BSAAs) for the treatment of COVID-19 and other emerging infectious diseases. There are no effective therapeutic agents currently available in the fight against the global COVID-19 pandemic, in which SARS-CoV-2 has infected millions of people in confirmed cases and caused hundreds of thousands of fatalities. Drugs that target a single virus, like the inhibitors of HIV reverse transcriptase and influenza neuraminidase, require a comprehensive understanding of the lifecycle and disease mechanisms of the virus, which makes development of these drugs necessarily time-consuming. Outbreaks of infection caused by novel emerging highly pathogenic viruses, including avian influenza, SARS, Ebola, Zika virus (ZIKV) and SARS-CoV-2, have become a major concern in the past two decades. We cannot rely on the traditional virus-specific drugs to treat diseases caused by these unpredictable emerging viruses. Therefore, it is extremely important to develop BSAAs effective against a range of viruses. My laboratory has been studying anti-viral innate immune responses, particularly the IFN-I signaling pathway and its downstream gene program, for the last 20 years. While the field has previously focused only on interferon-stimulated genes (ISGs), we have demonstrated that ISGs like cholesterol 25- hydroxylase (CH25H) and IFN-I downregulated genes like fatty acid synthase (FASN) both play important roles in limiting viral infection and replication. We have also identified multiple small molecules for use as BSAAs, including 25-hydroxycholersterol (25HC), the metabolic product of CH25H, and the FASN inhibitor C75. In addition, we have extensively studied host innate immune responses to coronaviral infection: we have published multiple papers that explain how the host IFN-I signal transduction pathway is activated in response to infection by coronaviruses like murine hepatitis virus (MHV) and how coronaviruses can suppress their host’s innate immune responses through the viral papain-like protease (PLpro). Most importantly, in our preliminary studies we found that 25HC and C75 both have strong inhibitory effects against SARS-CoV-2 infection. We hypothesize that the IFN-I-mediated antiviral gene program involves not only upregulation of antiviral ISGs but also downregulation of the host genes required for viral infection and replication. We further hypothesize that by identifying the IFN-I-mediated antiviral gene program against SARS-CoV-2, we will be able to develop novel antiviral agents to combat COVID-19 and other emerging threats. In this proposal, we will first determine the IFN-I gene program in innate immune response to SARS-CoV-2. We will also develop 25HC, 25HC analogs and FASN inhibitors as novel antiviral agents against SARS-CoV-2. We believe our studies will not only determine the IFN-I-mediated antiviral gene program in host innate immune response to SARS-CoV-2 infection but also develop BSAAs to treat COVID-19 and other emerging infectious diseases.