Role of type I interferon signaling in Zika virus infection of the brain

I 型干扰素信号在寨卡病毒大脑感染中的作用

• 批准号: 9256709
• 负责人: Susan R Weiss
• 金额: $ 20.13万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-30 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9256709
• 关键词: A549; Aborted Fetus; Address; Adult; Affect; Africa; Amniotic Fluid; Animal Model; Antigens; Antiviral Agents; Antiviral Response; Asia; Autopsy; Birth; Blood; Brain; Brazil; CRISPR/Cas technology; Caribbean region; Cell Culture Techniques; Cells; Central America; Central Nervous System Infections; Coronavirus; Culicidae; Data; Detection; Development; Disease; Double-Stranded RNA; Embryo; Emergency Situation; Engineered Gene; Engineering; Epidemic; Equilibrium; Fetal Development; Fetus; Flavivirus; French Polynesia; Gene Expression; Genes; High-Throughput RNA Sequencing; Histocompatibility Testing; Host Defense; Human; Immune; Immune response; In Vitro; Infection; Insecta; Interferon Type I; Interferon-alpha; Interferons; Island; Knock-out; Knowledge; Learning; Length; Ligase; Liver; Mexico; Microcephaly; Microglia; Micronesia; Monkeys; Mothers; Mus; Neonatal; Nerve Degeneration; Neuraxis; Neuroglia; Neurologic; Neurons; Newborn Infant; Organ; Outcome; Pathology; Pathway interactions; Peripheral; Placenta; Play; Predisposition; Pregnancy; Pregnant Women; Primary Infection; Production; Proteins; RNA Sequences; Reporting; Ribonucleases; Role; Route; Signal Pathway; Signal Transduction; Skin; South America; Spleen; Testing; Therapeutic; Time; Tissues; Tropism; Uganda; Viral; Viral Genome; Virulence Factors; Virus; Virus Diseases; Virus Replication; West Nile virus; Wild Type Mouse; World Health Organization; Zika Virus; brain cell; brain tissue; cell type; design; eIF-2 Kinase; fetal; in vivo; information gathering; mortality; neonatal brain; neonate; nervous system disorder; neurotropic virus; nonhuman primate; oligoadenylate; pandemic disease; particle; pregnant; response; therapeutic development; transcriptome; transmission process; viral RNA

Zika virus (ZIKV) is an insect borne flavivirus which until recently was found within a narrow equatorial belt from Africa to Asia. ZIKV has emerged across the globe and has reached pandemic levels, spreading to Mexico, Central America, the Caribbean and South America. While ZIKV usually causes mild disease, infection of pregnant women has been suspected to cause a significant increase in babies born with microcephaly in Brazil, starting in late 2015, presumed to have become infected transplacentally a previously unknown route of infection. The association of ZIKV with abnormal brain development in the fetus has been supported by the recent detection of ZIKV RNA sequences and antigens along with neurodegeneration in aborted fetuses and babies that died soon after birth. Thus it is vital that we learn as much as possible of how ZIKV replicates and interacts with its host, including in cells of the CNS. The type I IFN response is an early potent antiviral response to viral invasion that can have a major impact on restricting virus spread as evidenced by significantly increased virus replication and rapid mortality as well as loss of organ tropism barriers observed in mice lacking type I IFN signaling. The CNS expresses low levels of interferon signaling genes compared to peripheral organs suggesting the CNS may be less prepared to restrict viral invasion. In addition, viruses suppress or evade the host immune response in order to facilitate their replication within a host and cause disease and therefore encode proteins with host IFN antagonist activities. The outcome of infection therefore depends on the balance of host immune defenses and virus encoded virulence factors and this can differ among viruses and tissue types. We have carried out extensive studies of IFN signaling responses to neurovirulent murine coronavirus in various cell types in vitro as well as in the CNS and the periphery of mice. We propose to use our expertise as well as information gathered, along with our recent data demonstrating robust replication of ZIKV in human A549 cells where it activates the OAS-RNase L pathway and in primary murine neuronal and glial cell cultures to test the hypothesis that cell type and tissue specific differences in the type I IFN response to ZIKV may contribute to susceptibility of the fetal brain to ZIKV-induced pathology. In Aim 1 we will investigate the IFN signaling pathways during ZIKV infected human A549 cells and CRIPSR/Cas9 engineered knockout A549 cells as well as primary murine neuronal and glial cell cultures. In Aim 2 we will investigate ZIKV replication and IFN response in vivo in embryonic and neonatal mice. Upon completion of these aims we will have gained an understanding of the type I IFN response to ZIKV in vitro in human cells as well as murine CNS cells. We will also know more about IFN signaling in the developing mouse embryo and spread of ZIKV from infected mothers. These two approaches combined will aid in understanding the impact of IFN signaling on ZIKV infection and spread and in the more long term identify targets for the development of therapeutic strategies to treat ZIKV-induced disease.

寨卡病毒（ZIKV）是一种昆虫传播的黄病毒，直到最近才在狭窄的赤道带内发现 从非洲到亚洲。ZIKV已经在地球仪出现，并已达到大流行水平，蔓延到 墨西哥、中美洲、加勒比和南美洲。虽然ZIKV通常会导致轻微的疾病， 孕妇的怀孕被怀疑会导致婴儿出生时患小头畸形症的显着增加， 巴西，从2015年底开始，据推测已经通过胎盘感染，这是一种以前未知的感染途径。 感染ZIKV与胎儿异常脑发育的关联已经得到了以下研究的支持： 最近在流产胎儿中检测到ZIKV RNA序列和抗原沿着神经变性， 出生后不久死亡的婴儿。因此，我们必须尽可能多地了解ZIKV如何复制， 与其宿主相互作用，包括在CNS细胞中。I型干扰素反应是早期有效的抗病毒药物 对病毒入侵的反应，这可能对限制病毒传播产生重大影响， 显著增加的病毒复制和快速死亡率以及器官嗜性屏障的丧失， 缺乏I型IFN信号的小鼠。CNS表达低水平的干扰素信号传导基因， 外周器官表明中枢神经系统可能准备不足，以限制病毒入侵。此外，病毒 抑制或逃避宿主免疫应答，以促进它们在宿主内的复制， 因此编码具有宿主IFN拮抗剂活性的蛋白质。因此，感染的结果 这取决于宿主免疫防御和病毒编码的毒力因子的平衡， 在病毒和组织类型中。我们已经进行了广泛的研究IFN信号反应， 神经毒性鼠冠状病毒在体外的各种细胞类型以及在小鼠的CNS和外周中。 我们建议利用我们的专业知识以及收集到的信息，沿着我们最近的数据， ZIKV在人A549细胞中的稳健复制，其中它激活OAS-RNase L途径，并且在原代 小鼠神经元和神经胶质细胞培养物，以检验细胞类型和组织特异性差异在 对ZIKV的I型IFN应答可能有助于胎儿脑对ZIKV诱导的免疫应答的易感性。 病理在目的1中，我们将研究ZIKV感染的人A549细胞和人A549细胞中的IFN信号传导途径。 CRIPSR/Cas9工程化敲除A549细胞以及原代鼠神经元和神经胶质细胞培养物。在 目的2我们将研究ZIKV在胚胎和新生小鼠中的体内复制和IFN应答。后 完成这些目标后，我们将了解I型IFN对ZIKV的体外应答， 人细胞以及鼠CNS细胞。我们也将了解更多关于IFN信号在发育中的小鼠 ZIKV的胚胎和传播从受感染的母亲。这两种方法结合起来将有助于理解 IFN信号传导对ZIKV感染和传播的影响，并在更长期内确定ZIKV的靶点。 开发治疗ZIKV诱导的疾病的治疗策略。