Mechanisms of Dantrolene Neuroprotection in Alzheimer's Disease

丹曲林对阿尔茨海默病的神经保护机制

基本信息

批准号：
10227272
负责人：
HUAFENG WEI
金额：
$ 33.28万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10227272
关键词：
2019-nCoV ACE2 Address Affect Alzheimer&apos s Disease Alzheimer&apos s disease patient Apoptosis Binding COVID-19 COVID-19 patient COVID-19 treatment Calcineurin Cathepsin L Cathepsins Cell Death Cell Nucleus Cell membrane Cells Cessation of life Clinical Clinical Research Cytosol Dantrolene Data Dose Endocytosis Endoplasmic Reticulum Endosomes Epithelial Exposure to Fibroblasts Functional disorder Future Genetic Transcription Goals Grant Homeostasis Human Impairment Infection Kidney Knock-in Lysosomes Mediating Membrane Mitochondria Modeling Mutation Neurons Nuclear Pore Complex Oxygen Oxygen Consumption Parents Pathologic Pathway interactions Patients Pharmaceutical Preparations Pharmacotherapy Play Process Production Proteins RNA replication Reactive Oxygen Species Reporting Risk Factors Role Ryanodine Receptor Calcium Release Channel Ryanodine Receptors SARS-CoV-2 infection SARS-CoV-2 inhibitor SARS-CoV-2 spike protein Skin Stress Testing Toxic effect Transfection Virus Virus Diseases Virus Replication aged cell injury cytokine cytokine release syndrome drug repurposing efficacy evaluation endoplasmic reticulum stress experimental study induced pluripotent stem cell inhibitor/antagonist neuroblastoma cell neurogenesis neuroprotection nuclear factors of activated T-cells pandemic disease presenilin-1 prevent receptor sensor side effect stem cells synaptogenesis translational study viral RNA

项目摘要

Abstract Coronavirus disease 2019 (COVID-19), a pandemic affecting millions of patients around the world, is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with no effective drug treatments. Both infection and transfection of virus in host cells require abnormally elevated Ca2+ concentrations in cytosol and endosome via overactivation of ryanodine receptors (RyRs) Ca2+ channel located on the membrane of the endoplasmic reticulum (ER). Dantrolene, an antagonist of RyRs, has been demonstrated to inhibit SARS- CoV-2-mediated host cell toxicity and damage. In the presence of RyRs overactivation and excessive and abnormal elevation of cytosolic and endosome Ca2+ concentrations, the spike proteins (S1, S2) of the virus play important roles in binding, fusion, and virus replication in the host cell, eventually leading to cell damage or death. Our long-term goal is to examine the efficacy and mechanisms of dantrolene to treat AD. The overall objective of this study is to investigate the effects and underlying mechanisms of dantrolene to protect against host cell damage or death induced by SARS-CoV-2 spike proteins-mediated overactivation of RyRs and associated Ca2+ dysregulation. Our central hypothesis supported by preliminary data is that dantrolene inhibits SARS-CoV-2 spike proteins-mediated cell damage by inhibiting the overactivation of RyRs and by restoring intracellular Ca2+ homeostasis in AD cells. We will test this hypothesis with the following specific aims. Specific Aim 1 (SA1). To determine the effects of dantrolene on SARS-CoV-2 spike (S) proteins-mediated Ca2+ dysregulation and cell damage by apoptosis in AD cells using induced pluripotent stem cells (iPSC), from skin fibroblasts of patients with either sporadic (SAD) or familial (FAD), and human neuroblastoma cells (SH-SY5Y), with knocked-in AD presenilin 1 mutation (M146L). Specific Aim 2 (SA2). To determine the effects and mechanisms of dantrolene on SARS-CoV-2 spike (S) proteins-mediated pathological pathways leading to host cell damage. We will examine the effects of S proteins on mitochondria oxygen consumption, ATP production, reactive oxygen species (ROS) production, ER stress, and cytokine release. We will correlate the results between SA1 and SA2. We expect that dantrolene will inhibit S proteins-mediated overactivation of RyRs and associated disruption of intracellular Ca2+ homeostasis and cell death by apoptosis.

抽象的 2019 年冠状病毒病 (COVID-19)，一种影响数百万患者的大流行病世界范围内，由严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 引起，没有有效的药物治疗。病毒在宿主细胞内的感染和转染都需要通过过度激活细胞质和内体中的 Ca2+ 浓度异常升高兰尼碱受体 (RyRs) Ca2+ 通道位于内质膜上网状结构（ER）。 Dantrolene 是 RyRs 的拮抗剂，已被证明可以抑制 SARS- CoV-2 介导的宿主细胞毒性和损伤。在 RyR 过度激活和胞浆和内体 Ca2+ 浓度过度且异常升高，峰值病毒的蛋白质（S1、S2）在病毒的结合、融合和复制中发挥着重要作用。宿主细胞，最终导致细胞损伤或死亡。我们的长期目标是检查丹曲林治疗 AD 的疗效及机制。本研究的总体目标是研究丹曲林保护宿主细胞的作用和潜在机制 SARS-CoV-2 刺突蛋白介导的 RyR 过度激活引起的损伤或死亡相关的 Ca2+ 失调。初步数据支持的中心假设是丹曲林通过抑制 SARS-CoV-2 刺突蛋白介导的细胞损伤 RyRs 过度激活并恢复 AD 细胞的细胞内 Ca2+ 稳态。我们将通过以下具体目标来检验这一假设。具体目标 1 (SA1)。确定丹曲林对 SARS-CoV-2 刺突 (S) 蛋白介导的 Ca2+ 失调的影响使用诱导多能干细胞 (iPSC) 检测 AD 细胞凋亡造成的细胞损伤散发性（SAD）或家族性（FAD）患者的皮肤成纤维细胞以及人类神经母细胞瘤细胞 (SH-SY5Y)，具有敲入的 AD 早老素 1 突变 (M146L)。具体的目标 2 (SA2)。确定丹曲林对 SARS-CoV-2 的作用和机制刺突 (S) 蛋白介导的导致宿主细胞损伤的病理途径。我们将检查 S 蛋白对线粒体耗氧量、ATP 产生的影响，活性氧 (ROS) 产生、内质网应激和细胞因子释放。我们将关联 SA1 和 SA2 之间的结果。我们预计丹曲林会抑制 S 蛋白介导的 RyR 过度激活以及相关的细胞内 Ca2+ 稳态和细胞破坏因细胞凋亡而死亡。