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

项目摘要 2019新型冠状病毒感染已达到大流行状态，确诊病例超过1.25亿例， 造成100万人死亡，成为历史上最致命的流行病之一。它与严重的急性 呼吸系统综合征的症状，死亡率高。当SARS-CoV-2的S蛋白 与血管紧张素转换酶2（ACE 2）受体结合，通过该受体进入宿主细胞 (Kuba例如，2005; Walls等人，2020年）。ACE 2在肺中高度表达，在肺中表达程度较低。 其它器官如心脏、肾和肠（Bavishi等人，2020年），这说明了 肺部感染的患病率。对7300多万患者电子健康记录数据的分析 表明SUD患者感染COVID的风险更高，后果更严重- 19、尤其是美国人。阿片类药物使用障碍（OUD）患者的可能性是其他患者的2.4倍。 比可卡因使用障碍（1.6倍），酒精使用障碍（1.4倍）， 烟草使用障碍（吸烟或vaping; 1.3倍）。我们最近的初步数据显示，慢性阿片类药物 在阿片类物质滥用的小鼠模型中治疗导致ACE 2表达显著增加， 在这些动物的肺和大脑中的表达。这些数据表明，长期接触阿片类药物， 肺中ACE 2表达增加将增加对COVID 19感染的易感性， 表明很可能有神经侵入性在本附录中，我们将检验以下假设： 阿片类药物使用障碍将增加肺部感染COVID 19的风险， 与脑细胞中ACE 2表达增加导致的神经病理学后果相关。 目的1：我们将研究ACE 2在雄性和雌性大鼠小肠、肺和脑细胞中的表达。 长期用吗啡治疗的雌性小鼠。我们将进一步调查是否有滥用药物的情况 艾滋病毒的感染进一步加剧了感染和疾病进展。在目标2中，我们将研究使用人性化的 敲入小鼠ACE 2受体的小鼠模型，SARS-CoV 2刺突蛋白的感染性- 使用体外和体内靶细胞的假型GFP。在目标3中，我们将研究肠道的作用 微生物群在调节小肠、肺和脑中ACE 2表达水平中的作用。

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.