Interplay between AMPK and Hippo Signaling Regulates Ocular Antiviral Response to Zika virus infection

AMPK 和 Hippo 信号传导之间的相互作用调节眼部对寨卡病毒感染的抗病毒反应

基本信息

批准号：
10540701
负责人：
Vaithilingaraja Arumugaswami
金额：
$ 52.03万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10540701
关键词：
3-Dimensional 5&apos -AMP-activated protein kinase Ablation Address Affect Antiviral Response Atrophic Attenuated Biological Models Blood-Retinal Barrier Brain Diseases Cell Culture System Cell Density Cell Proliferation Cell physiology Cells Cessation of life Chemicals Cicatrix Code Cre lox recombination system Cytometry Cytoplasm Development Disease Progression Exhibits Eye Eye Abnormalities Eye Development Eye Injuries Eye diseases Fetal Development First Pregnancy Trimester Functional disorder Genes Genetic Histology Human Immune In Vitro Inflammatory Injury Innate Immune Response Interferon-beta Interferons Knock-out LATS1 gene Link Long-Term Effects Mediating Metabolic Metabolism Modeling Molecular Mus Ocular Pathology Organoids Outcome Measure Pathogenesis Pathogenicity Pathologic Pathway interactions Phosphorylation Phosphotransferases Physiological Pigments Predisposition Process Proteins RNA chemical synthesis Reflex action Regulation Reporting Resources Retina Retinal Diseases Role Second Pregnancy Trimester Series Signal Pathway Signal Transduction Stress Structure of retinal pigment epithelium TBK1 gene Teratogens Testing Tissues Tumor Suppressor Proteins Viral Viral Genome Virulence Factors Virus Virus Replication Vision Visual impairment ZIKA ZIKV disease ZIKV infection Zika Virus cell injury cell type conditioning design emerging human pathogen experimental study fetal fetal infection fundus imaging human pathogen in vivo Model induced pluripotent stem cell innate immune pathways insight knockout gene macula mouse model nervous system disorder novel organ growth pathogen pharmacologic primary outcome retinal progenitor cell sensor stem stem cells therapeutic target transmission process viral DNA

项目摘要

PROJECT SUMMARY: Zika virus (ZIKV) is a teratogenic human pathogen that causes congenital eye and brain diseases. Affected babies exhibit vision impairment and associated ocular pathology, including loss of foveal reflex and macular pigment mottling, chorioretinal scarring, and macular atrophy. ZIKV has become endemic and local transmissions in the USA have been reported previously. The long-term effects of structural damage on vision, as well as the pathogenic processes of congenital ZIKV eye diseases are beginning to be understood. The signaling pathways governing normal eye development, which are dysregulated during ZIKV infection, are not well characterized. We recently carried out a series of experiments by establishing a ZIKV infectious ocular cell culture system and mouse models to understand the structural and molecular perturbations. For successful replication, viruses have evolved various strategies to evade innate immune response as well as to enhance the availability of cellular metabolites required to meet the heightened energy demand for viral genome synthesis. We found that the AMPKα, a cellular master energy sensor, is activated in the ZIKV-infected retinal cells. Moreover, pharmacological activation of AMPK resulted in attenuated ZIKV replication. Another interesting finding is that the YAP/TAZ factors in the tumor suppressor Hippo/SWH signaling pathway were induced early on, but degraded at later stage of ZIKV infection in RPE cells. Silencing YAP/TAZ resulted in reduced ZIKV replication. Since the energy sensor AMPK and Hippo signaling pathways control key cellular processes, including host antiviral responses, it is critical to understand the fundamental mechanism of these two pathways deregulation. We hypothesize that ZIKV modulates AMPK and Hippo signaling pathways in ocular cells to 1) increase intracellular metabolic resources, and 2) inhibit TBK1 to antagonize antiviral defense. These molecular changes can be orchestrated through viral coded factors resulting in the pathogenesis of ocular cell injury. The following specific aims will be investigated. Aim 1 focuses on systematically evaluating the role of AMPK- Hippo signaling on regulating antiviral response to ZIKV infection in RPE cells. The cross talk between these pathways will be investigated at the YAP/TAZ level. Pharmacological activation/inhibition, and gene knockout approaches in RPE cells will be carried out. Aim 2 is designed to elucidate the effect of ZIKV on Hippo and AMPK signaling pathways during retinal development. Human iPSC-derived 3D-retinal cup organoids will be used to investigate the link between retinal development and ZIKV-mediated deregulation of these key pathways. The ZIKV-encoded virulence factors regulating these pathways will be characterized. Aim 3 is to determine the effect of RPE-specific ablation of AMPK, TBK1, and Hippo signaling on the pathogenesis of ZIKV-induced chorioretinal atrophy in mice. This proposed study would yield novel insights into the pathogenesis of ZIKV in ocular diseases and identification of potential therapeutic targets.

项目摘要：寨卡病毒（ZIKV）是一种致造先天性眼和大脑的致病性人类病原体疾病。受影响的婴儿展览视力障碍和相关的眼病理学，包括凹foveal的丧失反射和黄斑色素斑点，常年疤痕和黄斑萎缩。 zikv已成为内在和当地在美国的传输先前已有报道。结构损害的长期影响在视觉上，以及先天性ZIKV眼疾病的致病过程，开始被理解。在ZIKV感染期间，控制正常眼发育的信号通路是表现不佳。最近，我们通过建立ZIKV感染性眼进行了一系列实验细胞培养系统和小鼠模型，以了解结构和分子扰动。为了成功复制，病毒已经发展了各种策略，以逃避先天免疫反应，并增强满足病毒基因组合成能量需求的增强所需的细胞代谢物的可用性。我们发现在ZIKV感染的残留细胞中激活了AMPKα，一种细胞主能传感器。此外，AMPK的药物激活导致ZIKV复制减弱。另一个有趣的发现肿瘤抑制河马/SWH信号通路中的YAP/TAZ因子早早诱导开启，但在RPE细胞的ZIKV感染后期降解。沉默yap/taz导致zikv减少复制。由于能量传感器AMPK和HIPPO信号通路控制钥匙细胞过程，因此包括宿主抗病毒反应，了解这两种途径的基本机制至关重要放松管制。我们假设ZIKV调节眼细胞中的AMPK和HIPPO信号通路为1）增加细胞内代谢资源，2）抑制TBK1拮抗抗病毒防御。这些分子可以通过病毒编码因子来策划变化，从而导致眼部细胞损伤的发病机理。这将研究以下特定目标。 AIM 1专注于系统地评估AMPK-的作用河马信号传导控制RPE细胞中对ZIKV感染的抗病毒反应。之间的串口这些途径将在YAP/TAZ级别进行研究。药理激活/抑制作用和基因将在RPE细胞中进行敲除方法。 AIM 2旨在阐明ZIKV对视网膜发育过程中的河马和AMPK信号通路。人IPSC衍生的3D视视网膜杯器官将用于研究视网膜发育与ZIKV介导的放松管制之间的联系这些关键途径。控制这些途径的ZIKV编码的病毒因子将被表征。目的 3是确定AMPK，TBK1和HIPPO信号对RPE特异性消融的影响 ZIKV诱导的小鼠共性萎缩的发病机理。这项拟议的研究将产生新的见解 ZIKV在眼部疾病中的发病机理和潜在治疗靶标的鉴定。