Development of 3D Alveolar Tissue Models, CRISPR-editing and Microbiota-immune Response Assay Platforms for Deciphering Human Lung Immunity

开发 3D 肺泡组织模型、CRISPR 编辑和微生物免疫反应测定平台，以破译人类肺免疫

• 批准号: 10579873
• 负责人: Derya Unutmaz
• 金额: $ 57.32万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-05 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10579873
• 关键词: 3-Dimensional; Acetylation; Address; Age; Air; Alveolar; Animals; Antiviral Response; Bacteria; Biological Assay; CRISPR/Cas technology; Cancer Patient; Cell Communication; Cell Shape; Cell physiology; Cells; Cellular Assay; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Communities; Data; Dendritic Cells; Development; Engineering; Environment; Epigenetic Process; Epithelial Cells; Epithelium; Exposure to; Gender; Genes; Genetic; Genetic Engineering; Goals; Hematopoietic Stem Cell subsets; Hematopoietic stem cells; Human; Immune; Immune response; Immunity; Immunology; Immunology procedure; In Vitro; Infectious Agent; Laboratories; Lentivirus; Location; Lung; Lung diseases; Lung immune response; Lymphocyte; Macrophage; Malignant neoplasm of lung; Mammalian Cell; Mediating; Metabolic; Metabolic Pathway; Methylation; Microbe; Modeling; Modification; Molecular; Myelogenous; Myeloid Cells; Outcome; Pathway interactions; Physiological; Repression; Resources; Role; Structure of parenchyma of lung; System; T-Lymphocyte; Technology; Testing; The Jackson Laboratory; Tissue Model; Tissues; Transcriptional Activation; Upper respiratory tract; Viral; Viral Pathogenesis; Virus Diseases; bioprinting; cell type; data resource; design; human model; human tissue; immune function; immunoregulation; in vitro Model; innovation; insight; lung microbiota; microbial; microbiome; microbiota; novel strategies; particle; progenitor; programs; respiratory; respiratory microbiota; respiratory virus; response; screening; stem; technology development; tool; γδ T cells

PROJECT SUMMARY TECHNOLOGY DEVELOPMENT PROJECT The goal of the Technology Development project (Tech Dev) is to develop approaches, tools and assays that address the needs of The Jackson Laboratory Cooperative Center on Human Immunology (JAX CCHI) and that advance the capabilities of the scientific community to tackle questions regarding human lung immunity, human immune-microbiota interactions and basic mechanisms of immune cells. Major questions related to lung immune function remain unanswered—such as the cell-to-cell interactions between immune and lung epithelial cells that shape responses to foreign agents, or how the presence of microbiota in the airways or within lung compartments influences the pathogenesis of viral infections and other lung diseases. A significant technical barrier to studying human immune-lung dynamics is the sheer complexity of the human lung—which constantly filters airborne particles, infectious organisms and air through dynamic interactions between the lung epithelium and resident immune cells such as macrophages or dendritic cells. This complexity cannot be easily modeled in animal systems or using deceased human lung tissue. To surmount these challenges, Tech Dev will focus on three innovative human tissue platforms: 1) three-dimensional (3D) bioprinted models of the lung and upper respiratory environment for investigating the functional lung-immune interactome during exposure to viral or metabolic agents; 2) CRISPR/Cas9-based tools to genetically engineer primary human immune cell subsets, hematopoietic stem cells and/or lung epithelial progenitors to probe cell function; and 3) a functional in vitro platform for screening lung-resident microbiota and determining their impact on human lung immune responses. Each of these platforms addresses a specific unmet need in the application and will enable us, respectively, to study the human lung immunity within a dynamic and physiologically relevant microenvironment, to interrogate specific cell types and molecular pathways predicted to respond to viral infections, and to assess the impact of bacterial metabolites isolated from human airways on antiviral responses. Through these efforts, the JAX CCHI will be equipped to address previously inaccessible questions related to lung-immune dynamics, towards a more mechanistic understanding of lung immune function. Our Specific Aims are: Aim 1. Develop in vitro models of human lung tissue-immune interactions using 3D bioprinting. Aim 2: Optimize CRISPR-based genetic tools for use in engineering primary human immune and lung epithelial cells. Aim 3: Develop a functional immune assay platform to determine the immunomodulatory landscape of human lung and airway microbiota.

项目概要 技术开发项目 技术开发项目 (Tech Dev) 的目标是开发方法、工具和检测方法， 满足杰克逊实验室人类免疫学合作中心 (JAX CCHI) 的需求，并且 科学界解决人类肺部免疫、人类免疫等问题的能力不断提高 免疫-微生物群相互作用和免疫细胞的基本机制与肺免疫相关的主要问题。 功能仍未得到解答，例如免疫细胞和肺上皮细胞之间的细胞间相互作用 塑造对外来物质的反应，或者气道或肺室内微生物群的存在方式 影响病毒感染和其他肺部疾病的发病机制，这是研究的一个重大技术障碍。 人类免疫肺动力学是人类肺部的绝对复杂性——它不断过滤空气中的细菌 通过肺上皮和居民之间的动态相互作用来清除颗粒、传染性生物体和空气 巨噬细胞或树突状细胞等免疫细胞无法轻易在动物中建模。 为了克服这些挑战，技术开发人员将重点关注三个方面。 创新的人体组织平台：1）肺部和上呼吸道的三维（3D）生物打印模型 用于研究暴露于病毒或代谢期间功能性肺免疫相互作用组的环境 2) 基于 CRISPR/Cas9 的工具，用于对初级人类免疫细胞亚群、造血细胞进行基因改造 干细胞和/或肺上皮祖细胞来探测细胞功能；3) 功能性体外平台 筛查肺部微生物群并确定它们对人类肺部免疫反应的影响。 这些平台解决了应用程序中未满足的特定需求，并使我们能够分别研究 动态和生理相关微环境中的人肺免疫力，以询问特定的 预测对病毒感染做出反应的细胞类型和分子途径，并评估细菌的影响 通过这些努力，JAX CCHI 将能够从人类呼吸道中分离出抗病毒反应的代谢物。 能够解决以前无法解决的与肺免疫动态相关的问题，以实现更 对肺免疫功能的机制了解我们的具体目标是： 目标 1. 使用 3D 生物打印开发人肺组织-免疫相互作用的体外模型。 目标 2：优化基于 CRISPR 的遗传工具，用于工程初级人类免疫和肺上皮 细胞。 目标 3：开发功能性免疫检测平台以确定人类的免疫调节景观 肺和气道微生物群。