Role of Gut Microbiome- Brain Axis in Modulating CNS Inflammasomes in the Neuropathology Produced by Opioid Exposure and HIV

项目摘要 新冠肺炎感染已达到大流行状态，确诊病例超过1.25亿例，确诊病例2.75万例 数百万人死亡，使其成为历史上最致命的流行病之一。它与严重的急性 呼吸系统综合症症状，病死率高。SARS-CoV-2感染始于其S蛋白 与血管紧张素转换酶2(ACE2)受体结合，通过该受体进入宿主细胞 (Kuba等人，2005年；Walls等人，2020年)。血管紧张素转换酶2在肺组织中高表达，在 其他器官，如心脏、肾脏和肠道(Bavishi等人，2020)，这解释了 肺部感染的患病率。7300多万例患者电子病历数据分析 研究表明，患有肥皂泡的人感染COVID的风险更高，并遭受更严重的后果- 19尤其是在非裔美国人中。那些有阿片类药物使用障碍(OUD)的人有2.4倍的可能性 新冠肺炎患者比可卡因使用障碍患者(1.6倍)、酒精使用障碍患者(1.4倍)高，以及 烟草使用障碍(吸烟或吸电子烟；1.3次)。我们最近的初步数据显示，慢性阿片类药物 阿片类物质滥用小鼠模型的治疗导致ACE2表达显著增加 在这些动物的肺和脑中表达。这一数据表明，长期接触阿片类药物 肺组织中ACE2表达增加会增加对COVID19感染及其表达的易感性 在大脑中显示出很高的神经侵袭性。在本补充材料中，我们将检验这一假设 阿片类药物使用障碍将增加肺部感染COVID19的风险，并将 与脑细胞中ACE2表达增加所导致的神经病理后果有关。 在目标1中，我们将研究ACE2在雄性和雄性大鼠小肠、肺和脑细胞中的表达。 长期使用吗啡治疗的雌性小鼠。我们将进一步调查药物滥用是否在 艾滋病毒的传播进一步加剧了感染和疾病的发展。在目标2中，我们将使用人性化的 在小鼠模型中，小鼠的ACE2受体被敲入，SARS-CoV2刺突蛋白的传染性- 利用体外和体内靶细胞进行假型绿色荧光蛋白。在目标3中，我们将研究肠道的作用 微生物群在调节ACE2在小肠、肺和脑中的表达水平。

Project Summary Infection with Covid-19 has reached pandemic status with more than 125 million confirmed cases and 2.75 million deaths making it one of the most deadliest pandemics in history. It is associated with severe acute respiratory syndrome symptoms with high fatality rates. SARS-CoV-2 infection is initiated when its S-protein binds to the angiotensin-converting enzyme 2 (ACE2) receptor through which it gains entry into the host's cells (Kuba et al., 2005; Walls et al., 2020). ACE2 is highly expressed in the in the lungs and to a lesser degree in other organs, such as the heart, kidneys, and intestines (Bavishi et al., 2020), which explains the increased prevalence of lung infection. An analysis of electronic health record data from more than 73 million patients showed that people with SUDs are at higher risk of contracting and suffering worse consequences from COVID- 19 particularly among African Americans. Those with an opioid use disorder (OUD) were 2.4 times more likely to have COVID-19 than those with cocaine use disorder (1.6 times), alcohol use disorder (1.4 times), and tobacco use disorder (smoking or vaping; 1.3 times). Our recent preliminary data show that chronic opioid treatment in a mouse model of opioid substance abuse resulted in significant increase in the expression ACE2 expression in the lungs and brain of these animals. This data suggests that long term exposure to opioids by increasing ACE2 expression in the lung will increase the susceptibility to COVID 19 infection and its expression in the brain suggests high likely hood of neuro-invasiveness. In this supplement we will test the hypothesis that substance use disorders with opioids will increase the risk for COVID 19 infection in the lung and will be associated with neuropathological consequence as a consequence of increased ACE2 expression in brain cells. In Aim 1: We will investigate the expression of ACE2 in small intestine, lung and brain cells in both male and female mice that are chronically treated with morphine. We will further investigate if substance abuse in context of HIV further exacerbates infection and disease progression. In Aim 2; we will investigate using a humanized mouse model where the murine ACE2 receptor is knocked in, the infectivity of the SARS-CoV2 Spike Protein- Pseudotyped GFP using both in vitro and in vivo target cells. In Aim 3 we will investigate the role of the gut microbiome in modulating ACE2 expression levels in the small intestine, lung and brain.