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: 10113531
• 负责人: Derya Unutmaz
• 金额: $ 57.23万
• 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/10113531
• 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; Epithelial Cells; Exposure to; Gender; Genes; Genetic; Genetic Engineering; Goals; Hematopoietic stem cells; Human; Immune; Immune response; Immunity; Immunology; Immunology procedure; In Vitro; Infectious Agent; Laboratories; Lentivirus; Location; Lung; Lung diseases; Lymphocyte; 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; Viral; Viral Pathogenesis; Virus Diseases; base; bioprinting; cell type; data resource; design; human model; human tissue; immune function; immunoregulation; in vitro Model; innovation; insight; lung microbiota; macrophage; 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.

项目摘要技术开发项目 技术开发项目（Tech Dev）的目标是开发方法，工具和测定法 满足杰克逊实验室免疫学中心（JAX CCHI）的需求，并 提高科学界的能力来解决有关人类肺免疫学的问题 免疫微生物群的相互作用和免疫细胞的基本机制。与肺免疫有关的主要问题 功能仍未得到答复，例如免疫和肺上皮细胞之间的细胞间相互作用 形成对外国代理的反应，或在气道中或肺部室内的微生物群的存在 影响病毒感染和其他肺部疾病的发病机理。学习的重要技术障碍 人类免疫肺动力学是人类肺的纯粹复杂性，它不断过滤空中 通过肺上皮与居民之间的动态相互作用的颗粒，传染性生物和空气 免疫细胞，例如巨噬细胞或树突状细胞。这种复杂性不能轻易在动物中建模 系统或使用已故的人肺组织。为了克服这些挑战，技术开发人员将重点关注三个 创新的人体组织平台：1）肺和上呼吸道的三维（3D）生物打印模型 在暴露于病毒或代谢期间调查功能性肺免疫相互作用的环境 代理人； 2）基于CRISPR/CAS9的基于基因工程的工具，用于造血的基本人类免疫核管子集 干细胞和/或肺上皮祖细胞以探测细胞功能； 3）一个功能性的体外平台 筛查肺居民微生物群，并确定其对人肺免疫调查的影响。每个 这些平台满足了应用程序中的特定未满足需求，并将分别使我们能够研究 在动态和身体相关的微环境中的人类肺免疫学，以询问特定 细胞类型和分子途径预计会对病毒感染有反应，并评估细菌的影响 从人类气道中分离出的抗病毒反应的代谢产物。通过这些努力，JAX CCHI将是 能够解决与肺免疫动力有关的以前无法访问的问题， 对肺免疫功能的机械理解。我们的具体目的是： AIM 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。 AIM 2：优化基于CRISPR的基因工具用于工程初级人类免疫和肺上皮 细胞。 目标3：开发功能性免疫测定平台来确定人类的免疫调节景观 肺和气道微生物群。