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

摘要 这项提议的目标是利用芯片上的器官(器官芯片)在体外开发与临床相关的 可用于测试候选疗法疗效的人类流感感染模型，探索 不同患者群体的反应差异，并有可能开发针对以下目标的抗流感药物 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控培养 由平行生长的人体器官特异性组织细胞和血管内皮细胞排列的装置 微通道由细胞外基质涂层的多孔膜隔开。我们之前已经创建了 肺泡芯片以及由来自正常肺组织的细支气管上皮细胞排列的小气道芯片 捐赠者或患病患者，如慢性阻塞性肺疾病(COPD)患者，以及我们 表明它们真实地概括了活体观察到的人类病理生理学，包括肺部炎症。 此外，我们还创造了在体外代谢药物的人类肝脏芯片，以及 设计了一种用于多达10个人体器官芯片的自动化培养和流控耦合的仪器，最多可用于4个 周，这可以用来连接不同的器官芯片以一种生理的方式。重要的是，在预赛中 研究表明，我们成功地用甲型H1N1流感病毒(IAV)感染了这些细支气管上皮，证实了 宿主对感染反应的分子介体，并发现了一种潜在的新的抗病毒治疗方法 目标是这些调解人。在这个项目的UG3阶段，我们将演示肺气道和肺泡 从健康的人类捐献者或COPD患者分离的原代细胞排列的芯片可以用来模拟 先前在人类患者中观察到的IAV感染和相关肺部疾病的临床特征，包括 病毒复制和脱落，特征性炎性细胞因子的释放，循环的募集 免疫细胞和肺水肿，所有这些我们都将非侵入性测量。在UH3阶段，我们 将对现有抗病毒药物进行临床前疗效测试，并使用多组学分析和 生物信息学方法，以定义可翻译的生物标志物和确定新的潜在分子靶标。我们 也将利用这些洞察力来发现针对宿主反应的新的潜在疗法 感染，而不是病毒本身。我们的UG3的具体目标是：1)建立流感感染的模型 由健康供者和COPD患者的细胞排列的人肺通道和肺泡芯片重述 体内疾病反应，以及2)通过流体链接开发流感药物测试的综合模型 肺通道、肺泡芯片和肝脏芯片通过它们的血管通道。我们的UH3目标包括：1) 使用集成的器官芯片流感模型来衡量已知抗病毒疗法的有效性和安全性， 2)验证可翻译的流感感染生物标志物和使用 人体器官芯片模型与临床测量的比较，以及3)集成器官芯片的使用 流感模型，以确定针对宿主感染反应的新的抗病毒治疗药物。

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.