Real-Time Quantitation of Transport Across Vascular-Tissue Interfaces in Organ-On-Chip Models Using In Situ Mass Spectrometry

摘要 目前的体外平台对治疗药物的体内安全性、有效性和药代动力学的预测很差， 由于测试条件与生理条件相比存在显著差异。因此，毒品 毒性测试通常使用动物模型进行。然而，动物实验既昂贵又耗时。 在消费。此外，对使用动物的伦理担忧日益呼吁 减少/替换动物试验。为了克服这些挑战，生理相关的芯片上的器官 已经开发出了检测方法。这些分析模拟了药物输送过程中遇到的动态相互作用 并概括了生理流速、血管结构和组织(肝、肺、肾、 等)，从而提供改进的定量和预测能力，以通过 准确的毒性分析。然而，当前芯片上器官分析缺乏的关键组件之一是 实时分析化验中特定位置的药物浓度，以确定药物毒性 已定义的组织部位。 为了满足这一需求，我们建议将基于微流控技术的芯片上器官系统与芯片上质量相结合 光谱分析，以测量跨血管化肝脏结构的药物浓度。第一阶段的工作 将专注于将微流控设备与一种新的质谱仪(MS)分析相结合。此方法可启用 微流控装置中化学成分的在线时间和空间化学表征 这是第一次。ChemSitu方法使该方法能够连续采样和化学表征 在沿构造物的任意点直接从微流控装置直接产生小体积的液体 而不会负面地改变微流控系统的状态。一个多学科的科学家团队和 在基于微流体的细胞分析和仪器开发方面拥有专业知识的工程师 为顺利完成这一项目而组装。通过提供准确、定量和可预测的模型 开发的平台有望建立一种新的范式 用于体外评估对治疗药物的生理反应。

Abstract Current in vitro platforms are poor predictors of the in vivo safety, efficacy and pharmacokinetics of therapeutics, owing to a significant difference in the test conditions compared to physiological conditions. Therefore, drug toxicity testing is routinely performed using animal models. However, animal testing is expensive and time consuming. In addition, ethical concerns about the use of animals are increasingly calling for reduction/replacement of animal tests. To overcome these challenges, physiologically relevant organ-on-chip assays have been developed. These assays mimic the dynamic interactions encountered during drug delivery and recapitulates physiological flow rates, vascular architecture and the 3D nature of tissue (liver, lung, kidney, etc.), thereby providing improved quantitative and predictive capabilities to guide the development of drugs via accurate toxicity analysis. However, one of the critical components lacking from current organ-on-chip assays is the real-time analysis of drug concentration at specified locations within the assay to determine drug toxicity at defined tissue sites. To address this need, we propose to integrate our microfluidics-based, organ-on-chip systems with on-chip mass spectrometry analysis to measure drug concentrations across a vascularized liver construct. The Phase I effort will focus on integration the microfluidic device with a novel mass spectrometry (MS) assay. This method enables online temporal and spatial chemical characterization of chemical constituents within microfluidic devices by MS for the first time. The ChemSitu approach enables the means to continuously sample and chemically characterize small volumes of liquid directly from a microfluidic device at any point along the construct in near real-time and without negatively altering the state of the microfluidic system. A multi-disciplinary team of scientists and engineers with expertise in microfluidics-based cell assays and instrumentation development has been assembled for successful completion of this project. By providing an accurate, quantitative and predictive model of and quantitation of physiological interactions, the developed platform promises to establish a new paradigm for in vitro assessment of the physiological response to therapeutics.