A Multifaceted Approach to Study Tissue and Cell Type Specific Molecular Mechanisms of the Host Response to Acute/Chronic Viral Infection

研究宿主对急性/慢性病毒感染反应的组织和细胞类型特异性分子机制的多方面方法

• 批准号: 9557555
• 负责人: Emmanuel Thomas
• 金额: $ 38.38万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-10 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9557555
• 关键词: 3-Dimensional; Acute; Address; Automobile Driving; Cells; Chronic; DNA; Development; Disease; Disease Progression; Drosophila genus; Epithelial; Epithelial Cells; Fat Body; Gene Expression; Gene Expression Profile; Goals; Human; Human body; Immune; Immune response; Immune signaling; Innate Immune Response; Innate Immune System; Interferon-alpha; Interferons; Invaded; Liver; Microfluidics; Modeling; Modification; Molecular; Morbidity - disease rate; Natural Immunity; Organ; Pathogenesis; Pathway interactions; Patients; Pattern; Pattern recognition receptor; Production; Property; RNA; Research; Signaling Molecule; Stem cells; TLR3 gene; Tissues; Transformed Cell Line; United States; Viral; Virus; Virus Diseases; Virus Replication; antimicrobial peptide; base; cell immortalization; cell type; chronic infection; epigenetic regulation; fighting; human disease; immortalized cell; improved; in vitro Model; invention; microbial; mortality; new therapeutic target; novel; novel strategies; pathogen; physiologic model; programs; response

Project Abstract Very few viruses are able to manifest as chronic infections in humans. The intrinsic innate immune response provides a first line of defense against invading viruses; however, in the case of chronic viral infections, these initial responses that were ineffective at controlling virus replication can then cause disease over many years due to chronic activation. In most organs, epithelial cells are some of the first cells to encounter viruses in the human body and innate immune responses in these cells are paramount to driving subsequent immune control. Interestingly, epithelial cells predominantly produce type III interferons (IFNs) in response to viral infection whereas immune cells produce Type II IFNs (γ) and Type I IFNs (α/β) are produced by most cells in the body. The mechanism underlying cell type and tissue specific expression of the type III IFNs are unknown and likely involve regulation of epigenetics modifications, gene expression of pattern recognition receptors and associated signaling molecules. In drosophila, the fat body is the primary innate immune organ producing antimicrobial peptides in response to pathogens. The human liver, equivalent to the drosophila fat body in terms of function, utilizes Type III IFN responses to fight viral infection and likely possesses other unique properties with respect to innate immunity when compared to other organs. We have developed novel and exciting in vitro models that utilize primary epithelial cells from several organs that have intact innate immune responses when compared to immortalized or transformed cell lines. We and others have shown that these cell types are of critical importance in the development of disease since they directly detect components of viral pathogens. We therefore assert that primary cells are the optimal model to use for studies on innate immunity and we propose a novel approach to study innate immunity based on the innate immune pathways that we have demonstrated to be important for microbial pathogenesis. In addition, we are developing novel physiologic models incorporating primary epithelial cells, stem cell-derived epithelial cells, 3-dimensional chip and microfluidic-based platforms. The use of stem cell-derived cells would facilitate the identification of changes in gene expression, which occur during differentiation, that contribute to the unique innate immune system in epithelial cells. The specific goals of this program are to functionally characterize the innate immune response, including the production of Type III IFNs, to multiple viral pathogen associated patterns, including both DNA and RNA sensing pathways, and to elucidate the underlying molecular mechanisms through which innate immunity manifests in epithelial cells using sophisticated in vitro models. In addition, tissue specific and developmental expression of specific innate immune signaling components including, TLR3, STING and IRF7, will be addressed as a mechanism underlying tissue specific responses. Completion of these studies would offer the most in depth characterization of innate immunity in epithelial and other cell- types while improving our understanding of its contribution to human disease in multiple organs.

项目摘要 很少有病毒能够在人类中表现为慢性感染。 提供了抵御病毒入侵的第一道防线；然而，在慢性病毒感染的情况下，这些 最初的反应无法有效控制病毒复制，随后可能会在多年内导致疾病 由于慢性激活，上皮细胞是最先遇到病毒的细胞之一。 人体和这些细胞中的先天免疫反应对于驱动后续免疫至关重要 实际上，上皮细胞主要产生 III 型干扰素 (IFN) 以应对病毒。 而免疫细胞则产生 II 型干扰素 (γ) 和 I 型干扰素 (α/β)，而免疫细胞则由大多数细胞产生 III 型干扰素的细胞类型和组织特异性表达的机制尚不清楚。 并可能涉及表观遗传学修饰的调节、模式识别受体的基因表达和 在果蝇中，脂肪体是产生相关信号分子的主要先天免疫器官。 抗菌肽对病原体有反应，人体肝脏相当于果蝇的脂肪体。 就功能而言，利用 III 型干扰素反应来对抗病毒感染，并可能拥有其他独特的功能 与其他器官相比，我们开发了新颖且具有先天免疫特性的药物。 令人兴奋的体外模型，利用来自具有完整先天免疫的多个器官的原代上皮细胞 我们和其他人已经表明，与永生化或转化的细胞系相比，这些细胞的反应。 类型在疾病的发展中至关重要，因为它们直接检测病毒的成分 因此，我们断言原代细胞是用于研究先天免疫的最佳模型。 我们提出了一种基于我们研究的先天免疫途径来研究先天免疫的新方法 必须证明其对于微生物发病机制的重要性此外，我们正在开发新的。 包含原代上皮细胞、干细胞来源的上皮细胞、3维芯片的生理模型 和基于微流体的平台的使用将有助于识别干细胞。 分化过程中发生的基因表达变化有助于形成独特的先天免疫系统 该计划的具体目标是在功能上表征先天细胞。 对多种病毒病原体相关模式的免疫反应，包括产生 III 型干扰素， 包括 DNA 和 RNA 传感途径，并阐明潜在的分子机制 通过使用复杂的体外模型，先天免疫在上皮细胞中表现出来。 特定先天免疫信号成分的组织特异性和发育表达，包括， TLR3、STING 和 IRF7 将作为组织特异性反应的基础机制来讨论。 这些研究将提供上皮细胞和其他细胞中先天免疫的最深入的表征。 类型，同时提高我们对其对人类多个器官疾病的贡献的理解。