Cardiotoxicity Assays on an Integrated Platform of a Heart-on-a-Chip and an Optical Immunosensor

摘要 基于芯片上人体器官平台的毒性分析对药物来说变得越来越重要 发现和发展，因为它们允许测试药物化合物对细胞的毒性作用 在进入昂贵的动物试验或临床之前，建立与生理相关的人体组织模型 审判。芯片上器官模型的多个生理生化参数必须连续 监测，以评估这些模型对药物治疗的反应。虽然荧光 检测已被广泛应用于生物检测，它需要向样品中添加荧光团， 这可能会干扰细胞活动，更严重的是，实时荧光几乎是不可能的 标记器官模型在药物毒性测试过程中不断分泌的生物标记物。 因此，荧光检测在这里不是一个可行的选择--直接检测或“无标记”检测是可行的 需要监测药物与有机化合物相互作用的动态过程，以获得详细的 关于暂时性以及延迟性或累积性药物效应的信息。建议的总体目标是 因此，研究是通过使用人体器官来解决这些具有挑战性的药物毒性分析问题。 芯片型号由自动化的、无标签的光学生物传感器系统监控，该系统允许实时、长时间 人体心脏组织模型对不同药物反应的项、敏感性和动力学分析 微环境。 为了实现这一目标，我们提出了一种独特的方法，该方法基于我们的专利无标记生物传感器 与先进的芯片器官技术相结合。我们的开放式微腔结构 生物传感器使传感器芯片与芯片上的心脏模型能够通过 自动化微流控平台，具有再生传感器表面的内置能力 在很长一段时间内进行持续的动力学研究。芯片上的心脏模型将使用以下技术开发 一种创新的3D生物打印方法，使用 人诱导多能干细胞(IPSCs)来源的心肌细胞。微流控灌流 具有同时电刺激和机械刺激的内置能力的生物反应器 构造用于维持有机化合物的长期功能。另一方面，中国经济的长期稳定 建议的生物传感器系统将使用负热膨胀材料进行显著增强 传感器芯片的制作。一组药物的心脏毒性将通过量化的方式进行现场评估 人类心脏模型分泌的生物标记物。从这个项目开发的技术将是 极具变革性，可能适用于其他器官，并导致未来的个性化筛查 精确医学的药物毒性、疗效和药代动力学。

Abstract Toxicity assays based on human organs-on-a-chip platforms have become increasingly important for drug discovery and development, since they allow testing cytotoxic effects of pharmaceutical compounds on the physiologically relevant human tissue models before moving forward to expensive animal testing or clinical trials. Multiple physiological and biochemical parameters of the organ-on-a-chip models must be continually monitored in order to assess the responses of these models to drug treatments. Although fluorescence detection has been widely adopted for bioassays, it requires the addition of fluorophores to the samples, which may disturb cellular activities and more seriously, it is practically impossible for real-time fluorescence labeling of the biomarkers that are constantly secreted by the organ models during drug toxicity testing. Thus, fluorescence detection is not a viable option here - direct detection, or “label-free” detection, is required for monitoring the dynamic process of drug interactions with organoids to obtain detailed information on transient as well as delayed or cumulative drug effects. The overarching goal of the proposed research is thus to address these challenging issues of drug toxicity assays by using a human organ-on-a- chip model monitored with an automated, label-free, optical biosensor system that allows for real-time, long- term, sensitive, and kinetic analyses of human cardiac tissue models in response to various drugs in their microenvironments. To accomplish this goal, we propose a unique approach that is based on our patented label-free biosensor in conjunction with advanced organ-on-a-chip technologies. The open-microcavity configuration of our biosensor enables synergistic integration of the sensor chip with a heart-on-a-chip model through an automated microfluidic platform, which has the built-in capability to regenerate the sensor surface for continual kinetic studies over extended periods of time. The heart-on-a-chip model will be developed using an innovative 3D bioprinting approach that produces functional biomimetic cardiac organoids using cardiomyocytes derived from human induced pluripotent stem cells (iPSCs). A microfluidic perfusion bioreactor with the built-in capacity for simultaneous electrical and mechanical stimulations will be constructed to maintain long-term functionality of the organoids. On the other hand, the long-term stability of the proposed biosensor system will be significantly enhanced using negative thermal expansion materials for fabrication of the sensor chip. The cardiotoxicity of a panel of drugs will be evaluated in situ via quantification of the biomarkers secreted by the human cardiac model. The technology developed from this project will be highly transformative, which may be applied for other organs and lead to future personalized screening of drug toxicities, efficacy, and pharmacokinetics for precision medicine.