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

基本信息

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
项目类别：
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

项目摘要

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) 是欧文综合征 (OS) 的分子原因，欧文综合征是最常见的染色体疾病全世界人类的异常。肺部疾病是发病的一个重要原因 OS 患者的死亡率。反复呼吸道感染在这些人群中尤其常见个人，并且常常危及生命。然而，尽管最近的研究报告免疫患有 OS 的个体的失调和干扰素过度活跃，我们在了解 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髓系白细胞的动员，是针对呼吸道病原体的急性免疫反应的关键介质。器官芯片是采用微芯片制造方法创建的仿生、微流体、细胞培养装置包含连续灌注的中空微通道，其中居住着活组织细胞，排列成模拟器官水平的生理学。通过概括多细胞结构，组织-组织界面、化学梯度、机械线索和身体的血管灌注，这些设备产生传统二维或三维文化系统。它们还能够实现高分辨率、实时成像和体外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高度创新和先进的、生理相关的器官-器官串扰模型，用于描述 OS 对肺稳态生理学的影响模拟临床观察到的 T21 引起的免疫失调。为此，我们将创建一个微流体集成的小鼠多器官系统，可复制骨髓（BM）-肺轴，使用从野生型 (WT) 和 Op(16)1/Yey 小鼠（OS 小鼠模型）分离的原代细胞。在同时，为了最终将研究结果转化为人类，我们将集中部分努力产生人肺气道上皮、血管内皮和造血干细胞诱导健康受试者和 OS 个体的多能干细胞来重建肺和骨髓集成多器官芯片系统中的组织。我们将利用这些基于小鼠和干细胞的研究 T21 如何影响肺气道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发展和先天免疫细胞动员以应对吸入的空气传播流感病毒的挑战颗粒。我们的中心假设是，这种动态的生命微系统可以重现先天的 OS 中的免疫失调，揭示了肺部对挑战的过度免疫反应吸入感染剂，并能够发现气道功能中以前未知的病理学在多器官生理联系系统的背景下。