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.

麻疹病毒(MV)是发展中国家儿童死亡的主要原因，尽管有可用的 减毒活疫苗已有40多年的历史。免疫功能严重受损的人尤其有可能患上 MV.MV感染的最严重表现，包括脑炎，发生在有损害的人身上。 细胞免疫。MV在多达一半的常规病例中影响中枢神经系统(CNS)； 充分的细胞免疫感染已被根除，但细胞免疫受损的个体仍处于 这是一个不利因素。 在最近在南非爆发的MV疫情中，有几人死于MV CNS感染。我们分析了 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)--已经出现。正常情况下，MV F被合成并 保持预融合状态，直到它到达细胞表面，而这种预融合状态本质上是 在需要来自H的信号的过程中，不稳定和热力学驱动到聚变后状态 H与受体的相互作用。然而，在“中枢神经系统适应”病毒的情况下，F基因的突变 使其能够在较少依赖H与两个已知MV细胞的相互作用的情况下促进融合 受体；这个F是独立于H或受体激活的。 CNS分离物F是识别阻止MV传播的小分子的理想靶点 在中枢神经系统中，通过破坏F蛋白的稳定，从而促进F蛋白的过早折叠到其融合后 州政府。这样的化合物将有效地减少可用于介导细胞- 与细胞的融合，最终阻止了MV在中枢神经系统的传播。我们已经修改并验证了一个基于 基于生物发光的高通量筛选(HTS)分析。哥伦比亚大学医学中心 HTS设施文库将筛选有效阻止F-F介导的融合的小分子 从中枢神经系统分离出来的。 目的1：进行小分子文库的初步高温超导实验。细胞系和细胞系中的正交筛选 人类神经元将证实对抗活病毒的有效性。 目标2：选择的小分子的作用机制将用特定的官能团进行评估 化验。体外疗效研究将评估小分子在相关组织中的抗病毒活性。病毒式传播 小分子选择压强下的进化研究将确定出现的可能性 病毒抵抗力。