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

项目概要/总体摘要 我们提议在杰克逊实验室 (JAX CCHI) 建立一个 U19 人类免疫学合作中心 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 人的肺。上皮屏障位于宿主和环境之间的界面，在那里它们感知入侵 病原体。树突状细胞 (DC) 将病原体衍生的抗原呈递给 T 细胞和 B 细胞以诱导免疫反应。 然而，人类肺组织环境对DC和其他细胞的影响，例如新发现的 先天淋巴细胞 (ILC) 家族以及细菌反应性 MAIT 细胞尚未完全了解。安 未充分研究的环境因素是肺部微生物组。众所周知，微生物群可以严格调节 免疫细胞的功能，特别是在粘膜表面，但这种情况在肺部如何发生尚未完全解决。 JAX CCHI 试图利用多学科实验来解决这些关键问题 将免疫学与上皮细胞生物学以及基因组、细胞、 在人体肺组织中鉴定出功能和微生物组参数。我们的总体假设是 粘膜 T 细胞对呼吸道病毒感染反应的质量和程度由交叉因素决定 微生物群、上皮细胞和白细胞之间的对话。为了解决这个假设，我们构建了 JAX CCHI 围绕两个综合研究项目，重点关注肺部抗病毒免疫的基本免疫机制； 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组免疫测定；用于存储和的样品核心 人体组织的分布；以及用于专门微生物组分析、培养和培养的微生物组核心 计算分析。我们的中心将汇集临床医生和肺免疫学专家， 微生物组、生物工程、基因组学和计算生物学，以实现我们的目标并最大化 这项研究的潜力。行政核心将为各部门提供协调、沟通和监督 程序。该 CCHI 的目标是： 1) 了解上皮细胞和免疫细胞的网络如何在 人肺调节对呼吸道病毒的先天和适应性免疫； 2) 定义炎症是如何驱动的 微生物组决定了组织的稳态，即免疫设定点； 3) 确定是否以及如何免疫 儿童哮喘和成人肺癌这两种肺部疾病的设定点发生了改变，这两种疾病对 对公众健康的影响； 4) 开发创新技术来模拟人类肺免疫动力学和 阐明肺部抗病毒反应的关键分子机制、细胞类型和途径。影响：通过 研究重点是人肺中抗病毒免疫的传感器、诱导剂和调节剂，我们的 CCHI 将 贡献见解，有助于改善起源于传染病和其他免疫疾病的结果 或继发影响肺部。

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.