MALDI-MS Imaging of Cells Exposed to 3D-Printed Fluidic Devices for PK/PD Studies

描述(由申请人提供)：这个项目的目标是开发一种强大的新方法来检查新药候选的药代动力学(PK)和药效学(PD)。使用3D打印的流体设备，细胞培养将被动态地添加治疗药物。3D打印设备比目前的技术有了实质性的改进，因为它含有多孔膜，可以同时给药和清除药物。在最初的发展阶段，3D结肠癌细胞培养，即所谓的球体，将用具有良好特征的化疗药物进行治疗。球体的分子变化将通过基质辅助激光/解吸电离成像质谱仪(MALDI-IMS)进行监测。由于药物及其代谢物都有确定的质量，这些物种的渗透和分布可以通过MALDI-IMS绘制在整个球体中。该项目对公众健康的好处在于有望提供一种强大的新工具，在非侵入性的、体外动态的背景下表征新药的PK/PD。这种方法将使研究人员有可能对新药新陈代谢背后的分子变化建立一个连贯的图景，从而有助于设计更有效的治疗方法，并改善患者的结果。该项目围绕三组活动构建。首先，将设计3D打印射流装置 并优化了3D细胞培养的剂量。完成的终端用户友好型设备将能够加载测试药物分子，并使用简单的梯度泵送方案操纵其清除半衰期。其次，球体的生长和剂量将在3D打印设备中进行优化。最后，在时间进程实验中，通过MALDI-IMS对球体进行剂量和成像。数据将通过主成分分析进行分析。作为一项初步的概念验证研究，球体将首先用特征良好的药物伊立替康治疗。进一步 研究将扩展到更复杂的鸡尾酒疗法，例如对临床方案FOLFIRI的简短模拟。

DESCRIPTION (provided by applicant): This project targets the development of a powerful new approach to examine the pharmacokinetics (PK) and pharmacodynamics (PD) of novel drug candidates. Using a 3D printed fluidic device, cell cultures will be dynamically dosed with therapeutics. The 3D printed device offers substantial improvements over current technology, as it contains porous membranes to allow both dosing and clearance of the drugs. In the initial development phase, 3D colon cancer cell cultures, known as spheroids, will be treated with well-characterized chemotherapies. Molecular changes to the spheroids will be monitored via Matrix Assisted Laser/Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS). As both the drugs and their metabolites have defined masses, the penetration and distribution of these species can be mapped throughout the spheroids with MALDI-IMS. The public health benefits of the project lie in the promise of a powerful new tool to characterize the PK/PD of new drugs in a non-invasive, dynamic in vitro context. This approach will make it possible for researchers to build a coherent picture of the molecular changes that underlie the metabolism of new drugs, thus helping to devise more effective treatments, and improve patient outcomes. The project is constructed around three sets of activities. First, the 3D printed fluidic devices will be designed and optimized to dose the 3D cell cultures. The completed end- user friendly device will enable loading of a test-drug molecule and manipulation of its clearance half-life using a simple gradient-pumping scheme. Second, growth and dosing of the spheroids will be optimized in the 3D printed device. Finally, spheroids will be dosed and imaging via MALDI-IMS in a time course experiment. Data will be analyzed via principal component analysis. As an initial proof-of-concept study, spheroids will first be treated with the well-characterized drug irinotecan. Further studies will expand to more complicated therapeutic cocktails, such as an abbreviated simulation of the clinical regime FOLFIRI.