In-vivo Drug Interaction of Cytochrome P450 probed by resonant enhanced Coherent anti-Stokes Raman Scattering (CARS) Microscopy

细胞色素P450是依赖于NAD(P)H，含6型血红素的单加氧酶，在人体内催化多种反应。它们提供了对抗外源药物的主要防御系统，它们参与了80%以上目前使用的药物的代谢，它们是当前癌症治疗的主要靶点。提出的工作包括在活细胞中可视化细胞色素P450活性的体内方法的发展。相干反斯托克斯拉曼散射显微镜是一种新颖而强大的工具，克服了传统振动光谱学的缺点。这种非线性技术利用振动对比，允许快速和非侵入性体内成像，具有高化学灵敏度和选择性以及高三维空间分辨率。该项目的目的是将该技术首次扩展到血红素蛋白领域，以便在分子水平上研究具有临床意义的药理学方面，如药物代谢，药物相互作用，酶-配体相互作用或癌症进化过程中发生的过程。这将作为不同参数的函数进行详细研究，例如不同药物或致癌物的浓度、氧含量和不同的细胞类型。此外，这项调查可以更好地了解与癌症演变有关的过程，例如，作为外部因素的结果，如药物或化学化合物。

Cytochromes P450 are NAD(P)H dependent, 6-type heme containing monooxygenases that catalyse a variety of reactions in the human organism. They provide the primary defensive system against xenobiotics, they are involved in the metabolism of more than 80% of all currently used drugs and they are main target in current cancer therapies. The proposed work comprises the development of an in-vivo method to visualize the activity of Cytochrome P450 in living cells. Coherent anti-Stokes Raman scattering microscopy is a novel and powerful tool that overcomes the disadvantages of conventional vibration Spectroscopy. This nonlinear technique utilizes vibrational contrast, which permits rapid and non-invasive in vivo imaging with high chemical sensitivity and selectivity as well as a high threedimensional spatial resolution. The aim of this project is to extend this technique for the first time into the field of heme proteins in order to study pharmacological aspects with clinical relevance on a molecular level, such as the metabolism of drugs, drug interactions, enzyme-ligand interactions or processes occurring during cancer evolution. This will be investigated in detail as a function of different parameters, such as concentrations of different drugs or carcinogens, oxygen content and different cell types. Moreover, the investigation could yield a better understanding of processes related to the evolution of cancer for examples as a consequence of outer factors, such as drugs or chemicals compounds.