Emulating Immune Dysregulation by Trisomy 21 in a Multi-Organ-on-a-Chip System

在多器官芯片系统中模拟 21 三体的免疫失调

• 批准号: 10292703
• 负责人: Kambez Hajipouran Benam
• 金额: $ 210.19万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10292703
• 关键词: 3-Dimensional; Acute; Adult; Affect; Animals; Architecture; Benchmarking; Biochemical Genetics; Biological; Biology; Biomimetics; Biopsy; Blood; Blood Vessels; Bone Marrow; Cell Culture Techniques; Cell Differentiation process; Cells; Chemicals; Chromosome 21; Chromosome abnormality; Clinical; Clinical Investigator; Collaborations; Cues; Data; Development; Devices; Dimensions; Down Syndrome; Endothelial Cells; Endothelium; Engineering; Environmental Exposure; Epithelial Cells; Exhibits; Exposure to; Future; Genetic Engineering; Genetic Materials; Goals; Hematology; Hematopoietic; Hematopoietic stem cells; Homeostasis; Human; Human Chromosomes; Hyperactivity; Hypersensitivity; Immune; Immune response; Immunobiology; In Vitro; Individual; Infection; Inflammation; Inhalation; Innate Immune Response; Interferons; International; Knowledge; Laboratories; Leukocytes; Life; Ligands; Link; Lung; Lung diseases; Lung infections; Mechanics; Mediating; Mediator of activation protein; Metabolic; Methods; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Morbidity - disease rate; Mus; Mutate; Myelogenous; Organ; Organ Model; Pathology; Pathway interactions; Perfusion; Physiological; Physiology; Poly I-C; Protocols documentation; Recurrence; Reporting; Resolution; Respiration Disorders; Respiratory Tract Infections; Sampling; Signal Transduction; Smoke; Structure of parenchyma of lung; Syndrome; System; TLR3 gene; Technology; Therapeutic; Time; Tissue Sample; Tissues; Translations; Validation; Vascular Endothelium; Virion; Virus Diseases; Work; airway epithelium; airway inflammation; base; human stem cells; human tissue; immune activation; in vivo; induced pluripotent stem cell; influenza infection; influenzavirus; innovation; intercellular communication; lung development; lung injury; microchip; microsystems; migration; model building; mortality; mouse model; organ on a chip; real-time images; reconstitution; respiratory pathogen; response; stem cell biology; stem cell model; stem cells; translational impact; two-dimensional

Trisomy 21 (T21) is the molecular cause of Oown syndrome (OS), the most common chromosomal abnormality in humans worldwide. Lung disorders represent an important cause of morbidity and mortality in people with OS. Recurrent respiratory infections are particularly common in these individuals and are often life-threatening. However, despite recent studies reporting immune dysregulation and interferon hyperactivity in individuals with OS, there is a critical gap in our understanding on how extra genetic material from chromosome 21 influences homeostatic immune activity of the lung, and innate immune activation and mobilization of myeloid leukocytes, which are key mediators of acute immune response, to respiratory pathogens. Organs-on-chips are biomimetic, microfluidic, cell culture devices created with microchip manufacturing methods that contain continuously perfused hollow microchannels inhabited by living tissue cells arranged to simulate organ-level physiology. By recapitulating the multicellular architectures, tissue-tissue interfaces, chemical gradients, mechanical cues, and vascular perfusion of the body, these devices produce levels of tissue and organ functionality not possible with conventional two- dimensional or three-dimensional culture systems. They also enable high-resolution, real-time imaging and in vitro analysis of biochemical, genetic and metabolic activities of living cells in a functional tissue and organ context. The overarching goal of this project is to apply microengineering principles of organ-on-chip technology and develop a highly innovative and advanced, physiologically relevant model of organ-organ crosstalk to delineate impact of OS on homeostatic physiology of the lung and emulate clinically observed immune dysregulation due to T21. For this, we will create a microfluidically integrated murine multi-organ system that reproduces bone marrow (BM)-lung axis, using primary cells isolated from wild-type (WT) and Op(16)1/Yey mice (a murine model of OS). In parallel, to enable eventual translation of findings to humans, we will focus part of our efforts in generating human lung airway epithelia, vascular endothelium and hematopoietic stem cells from induced pluripotent stem cells of healthy subjects and individuals with OS to recreate Lung and BM tissue in the integrated multi-organ chip system. We will utilize these murine and stem cell-based platforms to study how T21 affects normal functioning and biological responses of the lung airway epithelium and endothelium. Moreover, we will in real-time analyze inflammation development and innate immune cells mobilization in response to challenge with inhaled airborne influenza virus particles. Our central hypothesis is that this dynamic living microsystem can recapitulate innate immune dysregulation in OS, reveal a pulmonary exaggerated immune response to challenge with inhaled infective agents, and enable discovery of previously unknown pathologies in airway function in the context of a multi-organ physiologically linked system.

21三体（T21）是Oown综合征（OS）的分子原因，OS是最常见的染色体异常。 世界范围内的人类异常。肺部疾病是发病的重要原因， 复发性呼吸道感染在这些患者中特别常见。 人，往往是生命的威胁。然而，尽管最近的研究报告免疫 在OS患者中，调节异常和干扰素过度活跃，在我们的研究中有一个关键的差距。 了解来自21号染色体的额外遗传物质如何影响稳态免疫 肺的活动，以及先天免疫激活和骨髓白细胞的动员， 是呼吸道病原体急性免疫反应的关键介质。器官芯片是 用微芯片制造方法制造的仿生、微流体、细胞培养装置， 包含由活组织细胞占据的连续灌注的中空微通道， 模拟器官水平生理学。通过重现多细胞结构， 接口、化学梯度、机械提示和身体的血管灌注，这些设备 产生组织和器官功能的水平， 立体文化体系。它们还可以实现高分辨率、实时成像和体外成像。 分析功能组织中活细胞的生化、遗传和代谢活动 和器官背景。该项目的总体目标是应用微工程原理， 器官芯片技术，并开发一种高度创新和先进的，生理相关的 描述OS对肺稳态生理学影响的器官-器官串扰模型， 模拟临床上观察到的由于T21引起的免疫失调。为此，我们将创建一个 - 微流体整合的鼠多器官系统，其再现骨髓（BM）-肺轴， 使用分离自野生型（WT）和Op（16）1/Yey小鼠（OS的鼠模型）的原代细胞。在 与此同时，为了最终将发现转化为人类，我们将集中部分精力， 在产生人肺气道上皮细胞、血管内皮细胞和造血干细胞方面， 健康受试者和OS个体的诱导多能干细胞重建肺和BM 多器官集成芯片系统中的组织。我们将利用这些小鼠和干细胞为基础的 研究T21如何影响肺气道的正常功能和生物反应的平台 上皮和内皮。此外，我们将实时分析炎症的发展， 天然免疫细胞对吸入性空气传播流感病毒攻击的应答动员 粒子我们的中心假设是，这个动态的活的微系统可以重演先天的 OS中免疫失调，揭示了肺对 吸入的感染因子，并能够发现以前未知的气道功能的病理 在一个多器官生理连接系统的背景下。