Small molecules to block measles spreading in the central nervous system

小分子阻止麻疹在中枢神经系统中传播

• 批准号: 9986209
• 负责人: Matteo Porotto
• 金额: $ 49.26万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9986209
• 关键词: Academic Medical Centers; Affect; Antiviral Agents; Antiviral resistance; Attenuated Live Virus Vaccine; Biological Assay; Bioluminescence; Cell Line; Cell fusion; Cell membrane; Cell surface; Cells; Cellular Immunity; Central Nervous System Infections; Child; Child Mortality; Chimeric Proteins; Complex; Complication; Dependence; Developing Countries; Disadvantaged; Disease; Disease Outbreaks; Encephalitis; Evolution; General Population; Glycoproteins; HIV; Hemagglutinin; Home environment; Human; Immune; Immunity; Immunize; Immunocompromised Host; Impairment; Individual; Infant; Infection; Infectious Encephalitis; Lead; Libraries; Measles; Measles virus; Mediating; Membrane; Membrane Fusion; Modeling; Molecular; Molecular Conformation; Motor Neurons; Mutation; Neuraxis; Neurons; Paramyxovirus; Patients; Peptides; Periodicity; Persons; Population; Process; Receptor Cell; Recombinants; Reporting; Research; Resistance; Risk; SLAM protein; Signal Transduction; South Africa; Structure; Subacute Sclerosing Panencephalitis; Testing; Thermodynamics; Tissues; United States; Universities; Vaccinated; Vaccines; Viral; Viral Proteins; Virus; Virus Diseases; Vulnerable Populations; anti-viral efficacy; attenuated measles virus; base; brain tissue; combat; drug candidate; efficacy study; experimental study; high throughput screening; nectin; preference; premature; pressure; prevent; receptor; receptor binding; screening; small molecule; small molecule inhibitor; small molecule libraries; viral resistance; virus envelope

Measles virus (MV) is a leading cause of child mortality in developing countries despite the availability of a live attenuated vaccine for over 40 years. Severely immune-compromised people are particularly at risk for MV. The most serious manifestations of MV infection, including encephalitis, occur in people with impaired cellular immunity. MV affects the central nervous system (CNS) in up to half of routine cases; with adequate cellular immunity the infection is eradicated, but individuals with impaired cellular immunity are at a disadvantage. In a recent MV outbreak in South Africa several people died of MV CNS infection. We analyzed the viruses from these patients and found that specific intra-host evolution of the MV fusion machinery -- receptor binding protein (H) + fusion protein (F) -- had occurred. Normally, the MV F is synthesized and maintained in a pre-fusion state until it reaches the cell surface, and this pre-fusion state is intrinsically unstable and thermodynamically driven to the post-fusion state in a process that requires a signal from H upon H's interaction with receptor. However in the case of the “CNS-adapted” viruses, a mutation in F allows it to promote fusion with less dependence on interaction of H with the two known MV cellular receptors; this F is activated independently of H or receptor. The CNS isolate F represents an ideal target for identifying small molecules that block the spread of MV in the CNS by destabilizing the F protein, and thereby promoting premature folding of F to its post-fusion state. Such compounds will effectively decrease the amount of pre-fusion F that is available to mediate cell- to-cell fusion, ultimately halting spread of MV in the CNS. We have adapted and validated a cell based bioluminescence based High Throughput Screen (HTS) assay. The Columbia University Medical Center HTS facility library will be screened for small molecules that efficiently block the fusion mediated by the F from the CNS isolate. Aim 1: Primary HTS of small molecule libraries will be performed. Orthogonal screenings in cell lines and human neurons will confirm efficacy against live virus. Aim 2: The mechanism of action of selected small molecules will be assessed using specific functional assays. Ex vivo efficacy studies will assess the antiviral activity of small molecules in relevant tissues. Viral evolution studies under the selective pressure of small molecules will determine the potential for emergence of viral resistance.

尽管有可用的疫苗，但麻疹病毒 (MV) 仍是发展中国家儿童死亡的主要原因 减毒活疫苗已有 40 多年的历史。免疫力严重受损的人尤其面临感染的风险 MV。 MV 感染最严重的表现，包括脑炎，发生在功能受损的人群中 细胞免疫。在多达一半的常规病例中，MV 会影响中枢神经系统 (CNS)；和 有足够的细胞免疫，感染就会被根除，但细胞免疫受损的个体 一个缺点。 最近南非爆发的一次 MV 疫情中，多人死于 MV 中枢神经系统感染。我们分析了 来自这些患者的病毒，并发现 MV 融合机制的特定宿主内进化—— 受体结合蛋白（H）+融合蛋白（F）——已经发生。通常，MV F 被合成并且 保持在融合前状态直到到达细胞表面，这种融合前状态本质上是 在需要 H 信号的过程中不稳定且热力学驱动至聚变后状态 H 与受体相互作用。然而，在“适应中枢神经系统”的病毒中，F 发生突变 使其能够促进融合​​，而较少依赖 H 与两种已知 MV 细胞的相互作用 受体；该 F 的激活独立于 H 或受体。 CNS 分离株 F 是识别阻止 MV 传播的小分子的理想靶标 通过破坏 F 蛋白的稳定性，从而促进 F 过早折叠至融合后，从而在 CNS 中发挥作用 状态。此类化合物将有效减少可用于介导细胞-融合的预融合F的量。 细胞融合，最终阻止 MV 在中枢神经系统中的扩散。我们已经调整并验证了基于细胞的 基于生物发光的高通量筛选（HTS）测定。哥伦比亚大学医学中心 HTS 设施库将筛选可有效阻断 F 介导融合的小分子 从中枢神经系统中分离出来。 目标 1：将进行小分​​子文库的初级 HTS。细胞系的正交筛选和 人类神经元将证实对活病毒的功效。 目标 2：将使用特定功能评估选定小分子的作用机制 化验。离体功效研究将评估小分子在相关组织中的抗病毒活性。病毒性的 小分子选择压力下的进化研究将决定出现的潜力 病毒抵抗力。