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

描述（由申请人提供）：本项目旨在开发一种强大的新方法，以检查新药候选药物的药代动力学（PK）和药效学（PD）。使用3D打印的流体装置，细胞培养物将被动态地给予治疗剂。3D打印设备提供了对当前技术的实质性改进，因为它包含多孔膜以允许药物的给药和清除。在最初的开发阶段，3D结肠癌细胞培养物（称为球状体）将用充分表征的化疗药物进行治疗。将通过基质辅助激光/解吸电离成像质谱法（MALDI-IMS）监测球状体的分子变化。由于药物及其代谢物都具有确定的质量，因此可以使用MALDI-IMS在整个球体中绘制这些物质的渗透和分布。该项目的公共卫生益处在于有望提供一种强大的新工具，用于在非侵入性、动态体外环境中表征新药的PK/PD。这种方法将使研究人员能够建立一个连贯的图片的分子变化，基础上的代谢新的药物，从而帮助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展开。首先，将设计3D打印流体设备， 并优化了3D细胞培养物的剂量。完成的终端用户友好的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，球状体的生长和剂量将在3D打印设备中优化。最后，将在时间过程实验中通过MALDI-IMS对球状体进行给药和成像。将通过主成分分析对数据进行分析。作为一项初步的概念验证研究，球状体将首先用充分表征的药物伊立替康治疗。进一步 研究将扩展到更复杂的治疗鸡尾酒，如临床方案FOLFIRI的简化模拟。

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.