Tissue Chips for Toxicology Evaluation

描述（由申请人提供）：虽然化疗药物可以通过特定的细胞和分子分析来筛选其功效，但它在动物（人）中产生的毒性可以发生在多个靶向器官中。因此，虽然筛选潜在的化学药物库作为化疗药物可以加快，毒理学分析是耗时和昂贵的。这项提议的目标是使用来自小鼠多个器官的精确切割组织芯片同时检查化疗药物的体外毒性。这种对多种靶组织中毒性作用的快速检测将加快分析一种新的化疗药物的毒性潜力。组织芯片的优点是保留了器官的细胞结构，因此可以检测到特定部位的毒性作用。本建议的目标是：目标1。制定从精确切割的组织切片制备组织芯片的程序。在我们早期使用组织芯片的尝试中，我们在精确切割的组织切片上“盖章”组织芯片。我们计划进一步探索这种方法或研究其他技术。目标2。制定培养程序，在毒性试验所需的潜伏期内保持组织芯片的活力。我们在旋转摇床上孵育了较小的器官切片，使用底部带有头的多孔板来增加混合。这种方法可以很容易地适应于组织芯片。目标3。将毒性试验适用于与组织芯片相关的小生物量。我们之前已经对不同器官的组织切片进行了细胞毒性（坏死）、细胞凋亡、线粒体功能改变等检测。由于化疗药物有不同的毒性机制，我们需要进行一系列的分析来描述毒性。这些需要缩小规模，或者采用新的、更敏感的检测方法。将强调发展快速分析以加快毒性分析。目标4。用组织芯片验证现有化疗药物的毒性。这些研究将证明化疗药物的组织特异性，并验证组织芯片的选择性。目标5。概述正在开发的化疗药物的毒性。我们将与我们的癌症化疗药物发现小组合作，获得一系列先导化合物，并将利用组织芯片分析它们的毒性。我们的结果将与合同实验室获得的独立毒性谱进行比较，这些实验室也检查了这些化合物。这些研究应该验证我们是否已经开发出一种创新的方法来快速描述潜在化疗药物的毒性。

DESCRIPTION (provided by applicant): While chemotherapeutic agents can be screened for their efficacy by specific cellular and molecular analyses, the toxicity it can produce in an animal (man) can occur in multiple targeted organs. Thus, while screening of a library of potential chemicals as chemotherapeutic agents can be expedited, toxicological profiling is time-consuming and expensive. The goal of this proposal is to use precision-cut tissue chips from multiple organs from mice to simultaneously examine the in vitro toxicities of a chemotherapeutic agent. This rapid examination of toxic effects in multiple target tissues will expedite profiling of the toxic potential of a new chemo-therapeutic agents. Tissue chips have the advantage that the cellular architecture of the organ is retained so site-specific toxic effects can be detected. The Aims of this Proposal are: Aim 1. Develop the procedures for preparing tissue chips from precision-cut tissue slices. In our earlier attempts to use tissue chips, we "stamped" the tissue chips from our precision-cut tissue slices. We plan to further explore this method or examine other techniques. Aim 2. Develop incubation procedures for maintaining the viability of tissue chips for incubation periods required for toxicity testing. We have incubated smaller organ sections on a rotating shaker using multiwell plates with a bead at the bottom to increase the mixing. This approach can be readily adapted to the tissue chips. Aim 3. Adapt toxicity assays to the small biomass associated with the tissue chip. We have previously adapted assays for cytotoxicity (necrosis), apoptosis, altered mitochondrial function, etc for our tissue slices from different organs. Since the chemotherapeutic agents have different mechanisms of toxicity, we will need to have a battery of analyses to characterize the toxicity. These need to be scaled down or new, more sensitive assays applied. Development of rapid analyses to expedite profiling of the toxicity will be emphasized. Aim 4. Validate the toxicity of existing chemotherapeutic agents with the tissue chips. These studies will demonstrated the tissue specificity of the chemotherapeutic agents and validate the selectivity of the tissue chips. Aim 5. Profile the toxicity of chemotherapeutic agents under development. We will coordinate with our cancer chemotherapeutic drug discovery group to obtain lead series of compounds and will profile their toxicities with the tissue chips. Our results will be compared to independent toxicity profiles obtained from contract laboratories also examining these compounds. These studies should validate if we have developed an innovative approach to rapidly profile the toxicity of potential chemotherapeutic agents.