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

摘要 基于人类器官芯片平台的毒性测定对于药物治疗变得越来越重要。 发现和发展，因为它们允许测试药物化合物对细胞的细胞毒性作用。 在进行昂贵的动物试验或临床试验之前， 审判芯片上器官模型的多个生理和生化参数必须是连续的， 监测以评估这些模型对药物治疗的反应。虽然荧光 检测已被广泛用于生物测定，它需要向样品中加入荧光团， 这可能干扰细胞活动，更严重的是，实际上不可能进行实时荧光 标记药物毒性试验期间器官模型持续分泌的生物标志物。 因此，荧光检测在这里不是可行的选择-直接检测或“无标记”检测是可行的。 需要监测药物与类器官相互作用的动态过程，以获得详细的 关于短暂以及延迟或累积药物作用的信息。拟议的《行动纲领》的总体目标 因此，研究是通过使用人体器官-对-a-来解决药物毒性测定的这些挑战性问题， 芯片模型监测与自动化，无标签，光学生物传感器系统，允许实时，长时间， 人类心脏组织模型对各种药物的反应的长期、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， 结合先进的器官芯片技术。我们的开放微腔结构 生物传感器能够使传感器芯片与芯片上的心脏模型协同集成， 自动化微流体平台，其具有再生传感器表面的内置能力， 在较长的时间段内进行连续的动力学研究。芯片上的心脏模型将使用 一种创新的3D生物打印方法，使用 在一些实施方案中，本发明涉及来源于人诱导多能干细胞（iPSC）的心肌细胞。微流控灌注 具有同时进行电刺激和机械刺激的内置能力的生物反应器将被 构建以维持类器官的长期功能性。另一方面，长期稳定 所提出的生物传感器系统将使用负热膨胀材料显著增强 传感器芯片的制作。一组药物的心脏毒性将通过定量进行原位评估 人类心脏模型分泌的生物标志物。该项目开发的技术将 高度变革性，这可能适用于其他器官，并导致未来的个性化筛选 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.