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

项目摘要/摘要总体 我们建议在杰克逊实验室建立一个U19人类免疫学合作中心(JAX CCHI)来 阐明先天免疫网络形成对呼吸道病毒感染的适应性免疫反应 人类的肺。上皮屏障位于宿主和环境的交界处，在那里它们感觉到入侵。 病原体。树突状细胞(DC)将病原体来源的抗原呈递给T和B细胞以诱导免疫反应。 然而，人类肺组织环境对DC等细胞的影响，如新发现的 先天淋巴样细胞(ILC)家族以及细菌反应性MAIT细胞尚不完全清楚。一个 未被充分研究的环境因素是肺微生物群。已知的微生物区系关键地调节了 免疫细胞的功能，特别是在粘膜表面，但这是如何发生在肺部并没有得到充分的解决。 JAX CCHI试图通过一个多学科的实验来解决这些关键问题 一种将免疫学与上皮细胞生物学以及基因组、细胞、 在人类肺组织中确定的功能和微生物组参数。我们最重要的假设是 粘膜T细胞对呼吸道病毒感染的反应的质量和大小由交叉决定。 微生物区系、上皮细胞和白细胞之间的对话。为了解决这一假设，我们构建了JAX CCHI 围绕肺抗病毒免疫的基本免疫学机制的两个综合研究项目； 一个技术开发项目，将利用3D生物打印、基因创建复杂的细胞模型 支持项目目标的编辑工具和微生物免疫分析；用于存储和 人体组织的分布；以及用于专门的微生物组剖析、培养和 计算分析。我们中心将把临床医生和肺部免疫学专家聚集在一起， 微生物组、生物工程、基因组学和计算生物学来实现我们的目标并最大限度地 这项研究的潜力。行政核心将提供协调、沟通和监督 程序。CCHI的目标是：1)了解上皮细胞和免疫细胞网络是如何在 人类肺调节对呼吸道病毒的先天和获得性免疫；2)定义炎症如何驱动 由微生物群决定组织的稳定状态，即免疫设置点；3)决定是否以及如何免疫 在儿童哮喘和成人肺癌这两种肺部疾病中，设定点会发生变化，这两种疾病的主要特点是 对公众健康的影响；以及4)开发创新技术来模拟人类肺免疫动力学和 阐明肺抗病毒反应的分子机制、细胞类型和途径。影响：通过 研究集中在人类肺部抗病毒免疫的传感器、诱导剂和调节剂上，我们的CCHI将 提供见解，帮助改善源于 或者二次冲击肺部。

PROJECT SUMMARY/ABSTRACT OVERALL We propose a U19 Cooperative Center on Human Immunology at The Jackson Laboratory (JAX CCHI) to elucidate the innate immune networks that shape adaptive immune responses to respiratory viral infections in the human lung. Epithelial barriers lie at the interface between host and environment, where they sense invading pathogen. Dendritic cells (DCs) present pathogen-derived antigens to T and B cells to induce immune responses. However, the impact of the human lung tissue environment on DC and other cells, such as the newly identified innate lymphoid cell (ILC) family, as well as bacteria-reactive MAIT cells, is not completely understood. An understudied environmental factor is the lung microbiome. Microbiota are known to critically modulate the function of immune cells, particularly at mucosal surfaces, but how this occurs in the lung is not fully addressed. The JAX CCHI seeks to address these critical questions using a multi-disciplinary experimental approach that will integrate immunology with epithelial cell biology along with genomic, cellular, functional and microbiome parameters identified in human lung tissues. Our overarching hypothesis is that the quality and magnitude of mucosal T cell responses to respiratory viral infections are determined by the cross- talk between microbiota, epithelial cells and leukocytes. To address this hypothesis, we structured the JAX CCHI around two integrated research projects focused on basic immunological mechanisms of lung antiviral immunity; a technology development project that will create sophisticated cellular models leveraging 3D bioprinting, gene editing tools and microbiome-immune assays to support project objectives; a sample core for storage and distribution of human tissues; and a microbiome core for specialized microbiome profiling, cultivation, and computational analysis. Our Center will bring together clinicians with experts in lung immunology, the microbiome, bioengineering, genomics and computational biology to achieve our goals and maximize the potential of this research. An administrative core will provide coordination, communication and oversight for the program. The goals of this CCHI are to: 1) Understand how the networks of epithelial cells and immune cells in the human lung regulate innate and adaptive immunity to respiratory viruses; 2) Define how inflammation driven by the microbiome dictates the steady state of tissue, i.e., immune set-point; 3) Determine if and how this immune set-point is altered in two pulmonary diseases, childhood asthma and adult lung cancer, which have a major impact on public health; and 4) Develop innovative technologies to model human lung-immune dynamics and elucidate molecular mechanisms, cell types and pathways key to lung antiviral responses. Impact: Through studies focused on the sensors, inducers and modulators of antiviral immunity in the human lung, our CCHI will contribute insights that could help improve outcomes for infectious and other immune diseases that originate in or secondarily impact the lung.