Liver-on-a-chip bioengineering to model systemic health responses to pollution exposure

背景：空气污染可导致肺、血管、心脏和大脑的慢性疾病。目前的法规旨在将颗粒物限制在2.5微米（PM2.5）以下。然而，这一部分在颗粒和化学品的组成方面各不相同，致病因子和毒性途径尚未确定。我们正在开发一种新的污染危害识别平台，以确定使用小鼠应激报告模型（邓迪）和相关的表观遗传和转录变化（爱丁堡，CMVM）对环境污染物的毒理学反应。在动物模型中确定的暴露允许比基于人群的研究更灵敏地检测分子机制，但对许多污染源的适应性较差。在与工程（CSE）合作，我们将设计一个器官组织培养芯片作为一种新的测定环境细胞应激和表观遗传变化。肝脏在代谢环境污染物中起着关键作用，并且可能是结局的主要决定因素。我们的目标是开发和评估一种基于肝细胞的方法来预测人类暴露的不良后果。项目建议书（CSE、MVM和外部资助者（如有）之间的TMF相互作用必须明显）：我们的相互作用将结合联合收割机独特的细胞应激报告细胞系，这些细胞系是基于干细胞和表观遗传学专业知识（Pennings（CS）、Meehan（MGPH）、CMVM）和小鼠遗传学和毒理学（Wolf，邓迪）开发的，具有微流体器官芯片技术和生物工程中的光谱学（Elfick，CSE）。联合研究将开发组件，使细胞适应3D培养，并使设备适应环境暴露研究。NERC NE/W 002086/1（tinyurl.com/2p8m7y5n）正在资助我们的小鼠报告模型中暴露应激途径和表观遗传变化的鉴定，这些模型具有氧化应激，DNA损伤和CAR或AhR介导的外源性暴露的发光检测。目标1将获得携带与亲代动物相同的报告基因的胚胎干细胞系。这些干细胞将成为衍生其他细胞类型的可再生来源，目标2采用肝细胞分化方案。Horizon正在资助肝细胞系3D共培养的器官芯片研究。该技术作为肝脏预测毒理学模型进行了测试（PMID 36473994）。目标3将报告细胞整合到这个设计中，活成像细胞，并收集细胞用于有或没有污染物暴露的表观遗传学和转录组学研究以及数据分析。这项博士研究将为肝脏芯片研究提供有关细胞应激和表观遗传变化的信息。结果将与NERC联盟的数据进行比较，以解决研究问题，即基于3D肝细胞的自主反应是否可以模拟肝脏对污染的全身健康反应。

Background: Air pollution aggravates chronic diseases of the lung, blood vessels, heart and brain. Current regulations aim to limit particulate matter up to 2.5 micron (PM2.5). However, this fraction varies in composition of particles and chemicals, and the causal agents and toxicity pathways have yet to be determined. We are developing a novel pollution hazard identification platform to determine the toxicological responses to environmental pollutants using mouse stress reporter models (Dundee) and associated epigenetic and transcription changes (Edinburgh, CMVM). Defined exposures in an animal model permit more sensitive detection of molecular mechanisms than population-based studies, but are less adaptable to many pollution sources. In collaboration with Engineering (CSE), we will design an organ tissue culture on-a-chip as a novel assay for environmental cell stress and epigenetic changes. The liver plays a key role in metabol- ising environmental pollutants and can be a major determinant in outcomes. Our Aim is to develop and evaluate a liver cell-based approach to predict human exposure adverse outcomes. Project Proposal (Tangible interaction between CSE, MVM and external funder (if any) must be evident): Our Interaction will combine unique cell stress reporter cell lines developed based on stem cell and epigenetics expertise (Pennings (CS), Meehan (MGPH), CMVM) and mouse genetics and toxicology (Wolf, Dundee), with microfluidic organ-on-a-chip technology and optical spectroscopy in bioengineering (Elfick, CSE). Joint research will develop components to adapt the cells to 3D culture, and adapt the device to environmental exposure studies. NERC NE/W002086/1 (tinyurl.com/2p8m7y5n) is funding identification of exposure stress pathways and epigenetic changes in our mouse reporter models with luminescence detection of oxidative stress, DNA damage, and CAR or AhR mediated xenobiotic exposures. Objective 1 will derive embryonic stem cell lines that carry the same reporters as the parental animals. These stem cells will be a renewable source for derivation of other cell types, with Objective 2 adopting a liver cell differentiation protocol. Horizon is funding organ-on-a-chip studies with 3D co-culture of liver cell lines. The technology was tested as a model for liver predictive toxicology (PMID36473994). Objective 3 will integrate the reporter cells into this design, live-image cells, and collect cells for epigenetics and transcriptomics studies with or without pollutant exposure, and data analysis. The PhD study will inform liver-chip research regarding cell stress and epigenetic changes. Outcomes will be compared to NERC consortium data to address the Research Question whether 3D liver cell-based autonomous responses can model the liver in systemic health responses to pollution.