PRECLINICAL DRUG AND BIOLOGIC TESTING

NANT临床前试验实验室的目标是促进在NANT内产生假设驱动的临床试验，并为NANT临床研究人员在I期试验中优先考虑潜在的新疗法和合理使用联合疗法提供指导。 以下具体目标将有助于实现我们的目标： 1)保持具有良好特性的人类神经母细胞瘤细胞系的主细胞库。 2)保持DIMSCAN仪器的工作状态，以用于涉及化疗药物和IL-6、可溶性IL-6、抗IL-6单抗CNTO 328、STAT3抑制剂他汀类(项目1)、NK细胞、抗IGF1R单抗(项目2)、PI3K抑制剂、Aurora Kinase A I型和II类抑制剂(项目3)以及NANT研究人员建议的药物和药物组合(项目4)的实验。 3)通过将肿瘤细胞与具有代表性的微环境细胞(例如，单核细胞、间充质细胞)共培养来开发“微环境”模型，以通过针对肿瘤细胞-微环境细胞相互作用(例如，抗IL-6单抗；来那度胺)的DIMSCAN疗法进行测试(与项目1和2合作)。 4)使用DIMSCAN进行体外细胞毒试验，为项目研究人员设计实验、指导使用DIMSCAN和数据分析提供帮助。 5)使用免疫受损小鼠的多药耐药人神经母细胞瘤皮下和播散性疾病异种移植模型，通过1)确定选定药物组合的最大耐受剂量(MTD)；2)评估肿瘤移植组织中MTD处药物活性的药效学证据；3)评估最有效的组合对肿瘤生长延迟和小鼠存活的影响，以评估选定组合的抗肿瘤活性。

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.