Kidney Microphysiological Analysis Platforms (MAP) to Explore SARS-CoV-2 Receptors and Inhibitors. A supplement to Parent Grant: Kidney Microphysiological Analysis Platforms (MAP) to Optimize Function

用于探索 SARS-CoV-2 受体和抑制剂的肾脏微生理分析平台 (MAP)。

• 批准号: 10179916
• 负责人: JOSEPH VINCENT BONVENTRE
• 金额: $ 25.15万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-25 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10179916
• 关键词: 2019-nCoV; A549; Affinity; Animals; Anti-Inflammatory Agents; Apoptotic; Bedside Testings; Binding; Binding Proteins; Biological Assay; Biomimetics; Bleomycin; COVID-19; Cell Adhesion; Cells; Dengue; Dengue Virus; Devices; Diagnostic; Diagnostic tests; Disease; Ebola; Endocytosis; Endothelial Cells; Endothelium; Epithelial; Epithelial Cells; Epithelium; Evaluation; Exposure to; Extracellular Domain; Family member; Hepatitis A; Hepatocyte; Human; Immunoglobulins; In Vitro; Infection; Injury to Kidney; Interferometry; Interleukin-6; Kidney; Kidney Diseases; Kinetics; Label; Lateral; Ligands; Lung; Lung Adenocarcinoma; Lung diseases; Maintenance; Measurement; Measures; Mediating; Membrane Glycoproteins; Microfabrication; Microfluidics; Modeling; Monkeys; Mucins; Non-Small-Cell Lung Carcinoma; Peptidyl-Dipeptidase A; Phagocytosis; Physiological; Production; Proteins; Proximal Kidney Tubules; Renal tubule structure; Reporting; Role; Severe Acute Respiratory Syndrome; System; T-Lymphocyte; Testing; Therapeutic Agents; Time; Triplet Multiple Birth; Variant; Virosomes; Virus; Virus Receptors; Work; airway epithelium; base; bronchial epithelium; carcinoembryonic antigen-related cell adhesion molecules; coronavirus receptor; cytokine release syndrome; hepatitis A virus receptor; inhibitor/antagonist; kidney cell; nanodisk; nanoplasmonic; new technology; novel; novel diagnostics; oxidized lipid; pandemic disease; paracrine; parent grant; phosphatidylserine receptor; point-of-care diagnostics; prevent; prophylactic; rat KIM-1 protein; receptor; response; side effect; small molecule; small molecule inhibitor; stem cell biology; therapeutic candidate; uptake

Abstract Coronavirus disease 2019 (COVID-19) has reached pandemic proportions. Pulmonary and kidney disease are highly prevalent serious consequences of infection with SARS-CoV-2. Kidney Injury Molecule-1 (KIM-1) was identified by Drs Bonventre and Ichimura as the most upregulated protein in the injured kidney proximal tubule. KIM-1, also called TIM-1, is a receptor for hepatitis A, Ebola, Dengue and possibly SARS-CoV1 viruses. We hypothesize that KIM-1 is a receptor for SARS-CoV-2 both in renal tubule epithelial cells and in airway epithelial cells and that JB1, our newly discovered small molecule inhibitor of KIM-1, and/or nanodisc- incorporated KIM-1 ectodomain can be prophylactic and therapeutic agents for COVID-19. We also hypothesize that we can use the high-affinity binding of KIM-1 and ACE2 to the virus to create novel diagnostic devices for the virus. In Specific Aim 1 we will characterize the role of KIM-1 in promoting SARS-CoV-2 entry into kidney and lung epithelia using kidney microphysiological analysis platforms (MAPs) on chip and develop an ultrasensitive chip for the high-throughput evaluation of potential SARS-CoV-2 binding inhibitors. KIM-1 and ACE2 mediated endocytosis of SARS-CoV-2 biomimetic viruses (virosomes) will be compared in kidney and lung epithelial cells. We will evaluate the effects of KIM-1-mediated spike proteins or biomimetic virus cellular adhesion and uptake on production of paracrine factors which activate endothelial cells using a kidney-lung MAP. In order to understand binding and/or uptake kinetics of SARS-CoV-2 and characterize potential inhibitors we will develop an ultrasensitive nanoplasmonic triplets-based rapid lateral flow diagnostic chip for a rapid and sensitive inhibition assay using the kidney-lung MAP. This approach can also be used for point of care diagnostic testing for the virus. In Specific Aim 2 we will evaluate the efficacy of JB1, soluble KIM-1 ectodomain and nanodisc-incorporated KIM-1 or ACE2 to inhibit binding and internalization of SARS-CoV-2 biomimetic viruses by kidney and lung cells using an integrated lung-kidney MAP on chip. Potential inhibitors will be tested to evaluate whether they compete with S-protein and/or biomimetic virus binding and reduce IL-6 production. Binding affinity and kinetics between KIM-1 variants or ACE2 either as free ectodomains or incorporated into nanodiscs and the Spike protein will be measured using MicroScale Thermophoresis (MST) and Biolayer Interferometry.

抽象的 2019 年冠状病毒病 (COVID-19) 已达到大流行的程度。肺和 肾脏疾病是 SARS-CoV-2 感染的高度流行的严重后果。肾 Bonventre 博士和 Ichimura 博士确定损伤分子 1 (KIM-1) 是上调程度最高的 受损肾近曲小管中的蛋白质。 KIM-1，也称为 TIM-1，是肝炎的受体 A、埃博拉病毒、登革热病毒以及可能的 SARS-CoV1 病毒。我们假设 KIM-1 是一种受体 肾小管上皮细胞和气道上皮细胞中的 SARS-CoV-2 JB1，我们新发现的 KIM-1 小分子抑制剂，和/或纳米盘- 掺入的 KIM-1 胞外域可以作为 COVID-19 的预防和治疗剂。 我们还假设我们可以利用 KIM-1 和 ACE2 与 病毒，为该病毒创建新型诊断设备。在具体目标 1 中，我们将描述 KIM-1 在利用肾脏促进 SARS-CoV-2 进入肾和肺上皮细胞中的作用 芯片上的微生理分析平台（MAP），并为微生理分析平台开发超灵敏芯片 潜在 SARS-CoV-2 结合抑制剂的高通量评估。 KIM-1 和 ACE2 将在肾脏中比较 SARS-CoV-2 仿生病毒（病毒体）介导的内吞作用 和肺上皮细胞。我们将评估 KIM-1 介导的刺突蛋白的作用或 仿生病毒细胞粘附和摄取旁分泌因子的产生，激活 使用肾肺 MAP 检测内皮细胞。为了了解结合和/或摄取动力学 SARS-CoV-2 的研究并表征潜在的抑制剂，我们将开发一种超灵敏的 基于纳米等离子体三联体的快速侧流诊断芯片，可实现快速、灵敏的抑制 使用肾肺 MAP 进行测定。该方法还可用于护理点诊断 测试病毒。在具体目标 2 中，我们将评估 JB1、可溶性 KIM-1 的功效 胞外域和纳米盘掺入 KIM-1 或 ACE2，以抑制结合和内化 使用集成的肺肾 MAP 通过肾脏和肺细胞产生 SARS-CoV-2 仿生病毒 在芯片上。将测试潜在的抑制剂以评估它们是否与 S-蛋白和/或 仿生病毒结合并减少 IL-6 的产生。 KIM-1 之间的结合亲和力和动力学 变体或 ACE2 作为游离胞外域或掺入纳米盘和刺突蛋白中 将使用微尺度热泳动 (MST) 和生物层干涉测量法进行测量。