Lung-on-a-Chip Disease Models for Efficacy Testing

总结 该提案的目标是使用器官芯片（器官芯片）开发临床相关的体外 人类流感感染模型，可用于测试候选疗法的功效，探索 不同患者群体的反应存在差异，并有可能开发出针对性的抗流感药物 宿主对感染的反应而不是病毒本身我们的器官芯片是双通道微流体培养 由平行生长的人类器官特异性组织细胞和血管内皮细胞内衬的装置 微通道由多孔细胞外基质包被的膜分隔。我们以前创造了 肺肺泡芯片以及小气道芯片，其由来自正常人或正常人的细支气管上皮细胞内衬。 供体或患病患者，例如患有慢性阻塞性肺病（COPD）的个体，并且我们 表明它们忠实地概括了在体内观察到的人类病理生理学，包括肺部炎症 此外，我们还创造了人体肝脏芯片，可以在体外代谢药物， 设计了一种仪器，用于自动培养和流体耦合多达10个人体器官芯片， 这可以用来以生理方式连接不同的器官芯片。重要的是，初步 研究中，我们成功地用H1N1甲型流感病毒（IAV）感染了这些细支气管上皮细胞， 宿主对感染反应的分子介质，并发现了一种潜在的新的抗病毒治疗剂， 针对这些中介。在该项目的UG3阶段，我们将证明肺气道和肺泡 由从人类健康供体或COPD患者分离的原代细胞内衬的芯片可用于建模 IAV感染的临床特征和先前在人类患者中观察到的相关肺部疾病，包括 病毒复制和脱落，特征性炎性细胞因子的释放，循环中 免疫细胞和肺水肿，所有这些我们都将非侵入性地测量。在第三阶段，我们 将对现有的抗病毒药物进行临床前疗效测试，并使用多组学分析， 生物信息学方法来定义可翻译的生物标志物并识别新的潜在分子靶点。我们 还将利用这些见解来发现新的潜在疗法，针对宿主的反应， 感染，而不是病毒本身。我们的UG3具体目标是：1）建立流感感染模型， 人类肺气道和肺泡芯片，内衬来自健康供体和COPD患者的细胞， 体内疾病反应，以及2）通过流体连接开发用于流感药物测试的集成模型 肺气道、肺泡芯片和肝芯片通过其血管通道。我们的UH3目标包括：1） 使用集成的器官芯片流感模型来测量已知抗病毒疗法的功效和安全性， 2)验证流感感染的可翻译生物标志物和使用 通过与人类临床测量结果进行比较的器官芯片模型，以及3）使用集成的器官芯片 流感模型，以确定新的抗病毒治疗的目标宿主对感染的反应。

SUMMARY The goal of this proposal is to use Organs-on-Chips (Organ Chips) to develop clinically relevant in vitro models of influenza infection in humans that can be used to test efficacy of candidate therapeutics, explore variation in responses in different patient populations, and potentially develop anti-influenza drugs that target the host response to infection, rather than the virus itself. Our Organ Chips are 2-channel microfluidic culture devices that are lined by human organ-specific tissue cells and vascular endothelium grown in parallel microchannels separated by a porous extracellular matrix-coated membrane. We have previously created Lung Alveolus Chips as well as Small Airway Chips lined by bronchiolar epithelial cells from either normal donors or diseased patients, such as individuals with chronic obstructive pulmonary disease (COPD), and we showed that they faithfully recapitulate human pathophysiology observed in vivo, including lung inflammation and pulmonary edema In addition, we have created human Liver Chips that metabolize drugs in vitro, and engineered an instrument for automated culture and fluidic coupling of up to 10 human organ chips for up to 4 weeks, which can be used to link different Organ Chips in a physiological way. Importantly, in preliminary studies, we successfully infected these bronchiolar epithelium with H1N1 influenza A virus (IAV), identified molecular mediators of the host response to infection, and discovered a potential new antiviral therapeutic that targets these mediators. In the UG3 phase of this project, we will demonstrate that Lung Airway and Alveolus Chips lined by primary cells isolated from human healthy donors or COPD patients can be used to model clinical features of IAV infection and related lung disorders previously observed in human patients, including viral replication and shedding, release of characteristic inflammatory cytokines, recruitment of circulating immune cells, and pulmonary edema, all of which we will measured non-invasively. During the UH3 phase, we will conduct preclinical efficacy testing of existing antiviral drugs and use multi-omics analysis and bioinformatics approaches to define translatable biomarkers and identify new potential molecular targets. We also will leverage these insights to discover new potential therapeutics that target the host response to infection, rather than the virus itself. Our UG3 Specific Aims are 1) to develop models of influenza infection in human Lung Airway and Alveolus Chips lined by cells from healthy donors and COPD patients that recapitulate in vivo disease responses, and 2) to develop an integrated model for influenza drug testing by fluidically linking Lung Airway, Lung Alveolus Chips, and Liver Chips via their vascular channels. Our UH3 Aims include: 1) to use the integrated Organ Chip influenza model to measure efficacy and safety of known antiviral therapeutics, 2) to validate translatable biomarkers for influenza infection and therapeutic responses identified using the Organ Chip model by comparison with clinical measures in humans, and 3) to use the integrated Organ Chip influenza model to identify new antiviral therapeutics that target host responses to infection.