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：
10322026
负责人：
Vaithilingaraja Arumugaswami
金额：
$ 50.47万
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/10322026
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 Development Disease 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 Response 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 Pharmacology Phosphorylation Phosphotransferases Physiological Pigments Predisposition Process Protein Kinase 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 Viral load measurement 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 primary outcome retinal progenitor cell sensor 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) 是一种致畸人类病原体，可导致先天性眼睛和大脑畸形受影响的婴儿表现出视力障碍和相关的眼部病变，包括黄斑中心凹丧失。反射和黄斑色素斑点、脉络膜视网膜疤痕和黄斑萎缩已成为地方病。先前已报道过美国当地的传播结构损坏的长期影响。人们开始了解 ZIKV 对视力的影响以及先天性 ZIKV 眼病的致病过程。控制正常眼睛发育的信号通路在 ZIKV 感染期间失调，我们最近通过建立 ZIKV 感染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。在复制过程中，病毒进化出了各种策略来逃避先天免疫反应并增强免疫反应。满足病毒基因组合成的营养能量需求所需的细胞代谢物的可用性。我们发现 AMPKα（一种细胞主能量传感器）在 ZIKV 感染的视网膜细胞中被激活。此外，AMPK 的药理学激活导致 ZIKV 复制减弱。发现肿瘤抑制因子 Hippo/SWH 信号通路中的 YAP/TAZ 因子在早期就被诱导上，但在 RPE 细胞中 ZIKV 感染的后期降解。沉默 YAP/TAZ 会导致 ZIKV 减少。由于能量传感器 AMPK 和 Hippo 信号通路控制关键的细胞过程，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我们勇敢地说，ZIKV 将眼细胞中的 AMPK 和 Hippo 信号通路调节为 1) 增加细胞内代谢资源，2) 抑制 TBK1 以拮抗抗病毒防御。变化可以通过病毒编码因子来协调，从而导致眼细胞损伤的发病机制。将研究以下具体目标：目标 1 侧重于系统评估 AMPK 的作用。 Hippo 信号调节 RPE 细胞对 ZIKV 感染的抗病毒反应。这些途径将在 YAP/TAZ 水平上进行研究。将在 RPE 细胞中进行敲除方法，目标 2 旨在阐明 ZIKV 对 RPE 细胞的影响。视网膜发育过程中的 Hippo 和 AMPK 信号通路。人类 iPSC 衍生的 3D 视网膜杯。类器官将用于研究视网膜发育与 ZIKV 介导的视网膜发育失调之间的联系这些关键途径的 ZIKV 编码的毒力因子调节这些途径将得到表征。图3是确定AMPK、TBK1和Hippo信号传导的RPE特异性消融对这项研究将产生新的见解。研究 ZIKV 在眼部疾病中的发病机制并确定潜在的治疗靶点。