Fetal Zika virus infection: role of the human placenta

胎儿寨卡病毒感染：人类胎盘的作用

• 批准号: 9265293
• 负责人: Eva Harris
• 金额: $ 19.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-11-18 至 2018-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9265293
• 关键词: ARHGEF5 gene; Address; Antibodies; Antibody titer measurement; Antibody-Dependent Enhancement; Asians; Binding; Biological Models; Blood Circulation; Blood Vessels; Brazil; Calcified; Cells; Centers for Disease Control and Prevention (U.S.); Central America; Chimera organism; Chorion; Chorionic villi; Clinical; Collaborations; Complex; Congenital Abnormality; Cytomegalovirus Infections; Cytopathology; Decidua; Decidual Cell; Dengue; Dengue Infection; Dengue Virus; Detection; Disease; Emergency Situation; Environment; Epidemic; Epidemiology; Epithelial Cells; Fc Receptor; Fetal Growth Retardation; Fetal Membranes; Fetus; Fibroblasts; Flavivirus; Gestational Age; Health; Human; Human Biology; Immune; Immune Sera; Immunoglobulin G; Immunology; Impairment; In Vitro; Individual; Infant; Infection; Joints; Knowledge; Link; Maternal antibody; Mediating; Membrane; Microcephaly; Modeling; Molecular; Monoclonal Antibodies; Mothers; Nicaragua; Nicaraguan; Pathogenesis; Pathology; Pattern; Placenta; Play; Population; Positioning Attribute; Pregnancy; Proteins; Receptor Protein-Tyrosine Kinases; Reporting; Research; Research Personnel; Risk Factors; Role; Route; Running; Serotyping; Severities; Skin; Spontaneous abortion; Stem cells; System; Testing; Therapeutic; Therapeutic Monoclonal Antibodies; Time; Tissues; Titrations; Variant; Villus; Virion; Virus; Virus Diseases; Virus Receptors; Zika Virus; amnion; antibody engineering; calcification; congenital infection; cross reactivity; cytotrophoblast; differential expression; experience; fetal; helicase; human monoclonal antibodies; inhibitor/antagonist; macrophage; mouse model; neutralizing antibody; neutralizing monoclonal antibodies; novel therapeutics; placental infection; prevent; prototype; receptor; receptor binding; receptor expression; small molecule; small molecule inhibitor; stillbirth; transcytosis; translational impact; transmission process; trophoblast; viral RNA; viral transmission; virology; virus envelope

PROJECT SUMMARY/ABSTRACT The Zika epidemic that began in Brazil and has spread to Central America and US territories, caused by the Asian Zika virus (ZIKV) strain, has been reported by the CDC as definitively linked to severe birth defects – microcephaly, miscarriage and stillbirth. Detection of ZIKV RNA in the placenta and fetus and in maternal circulation for months, calcifications associated with long-standing infection, and intrauterine growth restriction indicate that virus-induced pathology impairs placental functions. To address the issue of transplacental ZIKV transmission, we propose a collaborative project between Dr. Lenore Pereira (UCSF) and Dr. Eva Harris (UC Berkeley), experienced investigators with complementary expertise in congenital infection of the human placenta and Flavivirus research, respectively. Our preliminary studies reveal that prototype and recently iso- lated Nicaraguan ZIKV strains infect cells in placental explants and primary cells isolated from human placenta that express AXL, Tyro3 and TIM1 tyrosine kinase receptors, which mediate ZIKV and the closely related dengue virus (DENV) infection in skin. Infected placental cells, including fetal amniotic epithelial cells, placental fibroblasts and trophoblast progenitor cells, developed cytopathology and expressed ZIKV envelope and non- structural NS3 proteins, and virus titers released depended on the receptors expressed and gestational age. Indicative of infection route, AXL was detected in decidua (uterine decidual cells and invasive cytotrophoblasts), chorionic villi (placental fibroblasts, Hofbauer cells and blood vessels) and fetal membranes (amniotic epithelial cells and trophoblast progenitor cells). Differential expression of receptors suggests how ZIKV could infect the decidua and spread to the placenta, fetus and amnion-chorion membranes. Further, in endemic regions, cross- reactive pre-existing antibodies to DENV could play a critical role in protection or pathogenesis of ZIKV in placenta tissues during pregnancy. Our overarching hypothesis is that placental primary cells and explants of decidua, chorionic villi and amnion-chorion membranes can be used as a model of the human placenta to define molecular mechanisms of ZIKV infection by free virions; the role of antibodies in neutralization, trans- cytosis, and enhancement of ZIKV infection in the placenta; and the translational impact of inhibitors and thera- peutic neutralizing monoclonal antibodies (MAbs). Our Specific Aims are: Aim 1. Identify cells at the uterine- placental interface that are susceptible to infection by Nicaraguan ZIKV strains in primary cells and placental explants, characterize cognate receptors, and identify small-molecule inhibitors of infection. Aim 2. Study the neutralizing and potentially enhancing role of ZIKV-specific and DENV-cross-reactive antibodies on infection and transcytosis in primary placental cells and explants and determine the therapeutic potential of neutralizing MAbs with modified Fc that preclude antibody-dependent enhancement. The proposed studies will reveal molecular mechanisms of ZIKV infection, routes of virus transmission to the fetus, and therapeutic potential for engineered antibodies and small molecule inhibitors that block infection and prevent congenital disease.

项目总结/摘要 寨卡病毒疫情始于巴西，并已蔓延到中美洲和美国领土， 亚洲寨卡病毒（ZIKV）株已被CDC报告为与严重出生缺陷明确相关- 小头畸形，流产和死胎胎盘和胎儿以及母体中ZIKV RNA的检测 循环数月，与长期感染相关的钙化，以及宫内生长受限 表明病毒引起的病理损害胎盘功能。为了解决经胎盘ZIKV的问题， 传输，我们建议Lenore佩雷拉博士（加州大学旧金山分校）和伊娃哈里斯博士（加州大学 Berkeley），经验丰富的研究人员，在人类先天性感染方面具有互补的专业知识， 胎盘和黄病毒研究。我们的初步研究显示，原型和最近的iso- 灭活的ZIKV毒株感染胎盘外植体中的细胞和从人胎盘分离的原代细胞 表达AXL，Tyro 3和TIM 1酪氨酸激酶受体，介导ZIKV和密切相关的 登革热病毒（DENV）感染皮肤。感染的胎盘细胞，包括胎儿羊膜上皮细胞、胎盘 成纤维细胞和滋养层祖细胞，发展细胞病理学并表达ZIKV包膜和非ZIKV包膜。 NS 3结构蛋白和释放的病毒滴度依赖于表达的受体和胎龄。 作为感染途径的指示，在蜕膜（子宫蜕膜细胞和侵袭性细胞滋养层）中检测到AXL， 绒毛膜绒毛（胎盘成纤维细胞、Hofbauer细胞和血管）和胎膜（羊膜上皮 细胞和滋养层祖细胞）。受体的差异表达表明ZIKV如何感染 蜕膜并扩散到胎盘、胎儿和胎盘绒毛膜。此外，在流行地区，交叉- 针对DENV的反应性预先存在的抗体可能在ZIKV的保护或发病机制中发挥关键作用 胎盘组织在怀孕期间。我们的总体假设是胎盘原代细胞和外植体 的蜕膜，绒毛膜绒毛和绒毛膜-绒毛膜可用作人胎盘的模型， 定义ZIKV通过游离病毒体感染的分子机制;抗体在中和中的作用，反式 ZIKV感染在胎盘中的增强;以及抑制剂和治疗的翻译影响。 中和性单克隆抗体（MAbs）。我们的具体目标是：目标1。识别子宫的细胞- 在原代细胞和胎盘界面中易受ZIKV毒株感染的胎盘界面， 外植体，表征同源受体，并鉴定感染的小分子抑制剂。目标2.研究 ZIKV特异性和DENV交叉反应性抗体对感染的中和和潜在增强作用 和转胞吞作用，并确定中和的治疗潜力。 具有排除抗体依赖性增强的修饰的Fc的MAb。这些研究将揭示 ZIKV感染的分子机制，病毒传播到胎儿的途径，以及ZIKV的治疗潜力。 工程抗体和小分子抑制剂，阻断感染和预防先天性疾病。