PRECLINICAL DRUG AND BIOLOGIC TESTING

NANT临床前测试实验室的目标是促进NANT内假设驱动的临床试验的生成，并为NANT临床研究者提供指导，以优先考虑I期试验的潜在新疗法，以及合理使用组合疗法。 以下具体目标将有助于实现我们的目标： 1)维持一个具有良好特征的人神经母细胞瘤细胞系的主细胞库。 2)维持DIMSCAN仪器处于工作状态，用于处理化疗药物和IL-6、可溶性IL-6、抗IL-6 mAb CNTO 328、STAT 3抑制剂stattic（项目1）、NK细胞、抗IGF 1 R mAb（项目2）、PI 3 K抑制剂、Aurora激酶A I型和II型抑制剂（项目3）以及NANT研究者建议的药物和药物组合（项目4）的实验。 3)通过将肿瘤细胞与代表性微环境细胞（例如，单核细胞，间充质细胞）以通过靶向肿瘤细胞-微环境细胞相互作用的DIMSCAN疗法（例如，抗IL-6 mAb;来那度胺）（与项目1和2合作）。 4)使用DIMSCAN进行体外细胞毒性试验，并协助项目研究人员设计实验，指导DIMSCAN的使用和数据分析。 5)使用免疫功能低下小鼠中的多药耐药人神经母细胞瘤的皮下和播散性疾病异种移植模型，通过以下方式评估选定组合的抗肿瘤活性：1）确定选定药物组合的最大耐受剂量（MTD）; 2）评估肿瘤异种移植组织中MTD时药物活性的药效学证据; 3）评估最具活性的组合对肿瘤生长延迟和小鼠存活的影响。

The objective of the NANT Pre-Clinical Testing Lab is to facilitate the generation of hypothesis driven clinical trials within the NANT, and to provide guidance to NANT clinical investigators in prioritizing potential new therapeutics for phase I trials, and rational use of combinatorial therapy. The following Specific Aims will aid in achieving our objective: 1) Maintain a master cell bank of well-characterized human neuroblastoma cell lines. 2) Maintain DIMSCAN instrument in working order for use for experiments dealing with chemotherapeutic agents and IL-6, soluble IL-6, anti-IL-6 mAb CNTO 328, the STAT3 inhibitor stattic (Project 1), NK cells, anti-IGF1R mAb (Project 2), PI3K inhibitors, Aurora Kinase A type I and II inhibitors (Project 3), and drugs and drug combinations suggested by NANT investigators (Project 4). 3) Develop a "microenvironment" model by co-culturing tumor cells with representative microenvironment cells (e.g., monocytes, mesenchymal cells) to test by DIMSCAN therapies targeting tumor cell - microenvironment cell interaction (e.g., anti-IL-6 mAb; lenalidomide) (collaboration with Projects 1 and 2). 4) Conduct in vitro cytotoxicity assays using DIMSCAN and provide assistance to project investigators in designing experiments, instructing in use of DIMSCAN and analyzing of data. 5) Assess anti-tumor activity of selected combinations using subcutaneous and disseminated disease xenograft models of multi-drug-resistant human neuroblastomas in immunocompromised mice by 1) determining maximal tolerated dose (MTD) of selected drug combinations; 2) assessing pharmacodynamic evidence of drug activity at the MTD in tumor xenograft tissue; 3) assessing the effect of the most active combinations on tumor growth delay and mouse survival.