Mechanistic characterization of SARS-CoV2 associated kidney injury

SARS-CoV 2是一种高度传染性的新型人类冠状病毒，可导致2019年冠状病毒病（COVID-19）。 19）。目前，美国有超过1650万人确认感染了SARS-CoV 2， 都死了重症COVID-19的特征是肺部和全身炎症以及多器官 严重的COVID患者中有很大一部分发生急性肾损伤。机制 SARS-CoV 2是如何引发这种严重的发病机制尚不清楚。最近的临床研究 表明细胞死亡，特别是诱导坏死性凋亡，可能是严重COVID-19的预测因子 疾病宿主限制坏死性凋亡的机制尚不清楚。初步数据显示， 干扰素诱导蛋白ISG 15作为坏死性凋亡的负调节因子， 病毒感染时的炎症反应。我们还表明，ISG 15缺陷小鼠迅速死亡， 以大量释放促炎细胞因子为特征的肾缺血-再灌注损伤。 在本提案中，我们将检验ISG 15作为调节细胞凋亡的关键宿主限制因子的假设。 肾脏内程序性坏死性凋亡和下游炎症反应，以限制急性肾损伤 在SARS-CoV 2感染期间。我们将利用来源于诱导多能干细胞的肾类器官， ISG 15已被CRISPR删除，CRISPR是一种与肾类器官和原代气管上皮共培养系统 培养物（hTECs）和SARS-CoV-2的体内小鼠模型，以提出几个问题，包括：1）SARS- CoV 2通过直接病毒转导或响应系统性免疫应答诱导肾上皮细胞损伤。 炎症？2)ISG 15是否调节坏死性细胞死亡以及促炎细胞因子/趋化因子 SARS-CoV 2感染过程中肾脏上皮细胞的产生？3)坏死性凋亡及其调节是否 ISG 15与SARS-CoV 2感染时急性肾损伤有关？总的来说，我们的研究将提供重要的 深入了解限制坏死性凋亡的宿主因素，可能是严重COVID-19的重要因素 导致肾损伤。

SARS-CoV2 is a highly contagious, novel human coronavirus that causes coronavirus disease 2019 (COVID- 19). Currently over 16.5 million people in the US have confirmed infection with SARS-CoV2 and over 300,000 have died. Severe COVID-19 is characterized by pulmonary and systemic inflammation and multi-organ dysfunction, with a significant portion of severe COVID patients developing acute kidney injury. The mechanism by which SARS-CoV2 triggers such severe pathogenesis is poorly understood. Recent clinical studies have suggested that cell death, especially the induction of necroptosis, may be a predictor of severe COVID-19 disease. The mechanism by which the host restricts necroptosis is unclear. In preliminary data we have shown that the interferon induced protein, ISG15, acts as a negative regulator of necroptosis and its downstream inflammatory responses during viral infection. We have also shown that ISG15 deficient mice rapidly succumb to ischemia-reperfusion injury of the kidney characterized by a massive release of proinflammatory cytokines. In this proposal we will test the hypothesis that ISG15 serves as a critical host restriction factor in regulating programmed necroptosis and downstream inflammatory responses within the kidney to limit acute kidney injury during SARS-CoV2 infection. We will utilize kidney organoids derived from induce pluripotent stem cells in which ISG15 has been deleted by CRISPR, a co-culture system with kidney organoids and primary tracheal epithelial cultures (hTECs), and in vivo mouse model of SARS-CoV-2 to ask several questions including: 1) Does SARS- CoV2 induce damage to kidney epithelial cells via direct viral transduction or in response to systemic inflammation?; 2) Does ISG15 modulate necroptotic cell death as well as proinflammatory cytokine/chemokine production in kidney epithelial cells during SARS-CoV2 infection?; 3) Does necroptosis and its regulation by ISG15 contribute to acute kidney injury during SARS-CoV2 infection? Overall, our studies will provide important insight into host factors that restrict necroptosis and could be an important contributor to severe COVID-19 induced kidney injury.