In vivo Drug Testing of Pediatric CNS Tumors Using Patient Derived Orthotopic Xenograft Models

使用患者来源的原位异种移植模型对儿童中枢神经系统肿瘤进行体内药物测试

• 批准号: 10300370
• 负责人: Xiaonan Li
• 金额: $ 63.44万
• 依托单位: LURIE CHILDREN'S HOSPITAL OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-14 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10300370
• 关键词: Animal Model; Animals; Autopsy; Biological Assay; Biological Models; Brain; Brain Neoplasms; Breeding; Cancer Etiology; Cancer Patient; Cells; Central Nervous System Neoplasms; Childhood; Childhood Brain Neoplasm; Childhood Central Nervous System Neoplasm; Childhood Glioblastoma; Clinic; Clinical; Clinical Drug Development; Clinical Trials; Cryopreservation; DNA; DNA methylation profiling; Databases; Diagnosis; Diffuse intrinsic pontine glioma; Dose; Drug Screening; Ensure; Ependymoma; Faculty; Funding Opportunities; Gene Expression; Glioblastoma; Glioma; Goals; Grant; Histopathology; Human; Implant; In Vitro; Injections; Institution; Investigation; Investigational Drugs; Laboratories; Location; Modeling; Molecular; Molecular Abnormality; Molecular Target; Mus; NOD/SCID mouse; New Agents; Operative Surgical Procedures; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Positioning Attribute; Preclinical Testing; Primary Neoplasm; RNA; Recurrent tumor; Relapse; Resistance; SCID Mice; Series; Serum-Free Culture Media; Specimen; System; Techniques; Testing; Time; Transplantation; Treatment Efficacy; Universities; Ursidae Family; Xenograft Model; Xenograft procedure; animal facility; base; cancer therapy; cell bank; cell killing; childhood cancer mortality; clinically relevant; cohort; cost effective; data mining; drug candidate; drug testing; efficacy evaluation; experience; implantation; in vivo; in vivo evaluation; medical schools; medulloblastoma; member; molecular modeling; molecular subtypes; mouse model; neoplastic cell; new therapeutic target; novel; novel anticancer drug; novel therapeutics; pre-clinical; programs; protein biomarkers; radioresistant; response; screening; success; therapeutic target; tumor; tumor xenograft; tumorigenic

This application is prepared in response to the funding opportunity: NCI Pediatric In Vivo Testing Program (U01), RFA-CA-20-034 to renew our existing PPTC UO1 grant. Specifically, we propose to continue the in vivo testing program for central nervous system (CNS) tumors using our panel of patient derived orthotopic xenograft (PDOX) models. Brain tumor is the leading cause of cancer-related death in children. One of the challenges in clinical drug development is how to effectively prioritize drug candidates to ensure clinical success in cancer patients. However, efforts in identifying new anti-cancer agents for that are most likely to be effective in the clinic have been blocked for many years due to the lack of clinically relevant and molecularly accurate model system. Fortunately, we have established a panel of 150 PDOX models of pediatric brain tumors through direct injection of patient tumor specimens into the brains of SCID mice. These PDOX models are shown to have replicated the histopathology and major genetic abnormalities of the original patient tumors even during serial sub-transplantations in vivo in mouse brains. They not only represent different clinical stage (i.e., at diagnosis, relapse and terminal/autopsy) but also replicate a broad spectrum of the newly identified molecular subtypes of nearly all types of pediatric brain tumors. The xenograft tumor cells can also be cryopreserved for sustained and on-demand supply of tumorigenic PDOX cells. This capacity combined with our optimized surgical procedure, with which we can implant up to 260 mice per day, makes it possible for us to test multiple (e.g., 6-10) drugs per year for every tumor type. Our objective is therefore to make use of this unique panel of PDOX models to examine therapeutic efficacy of new agents and to analyze mechanisms of action and therapy resistance in high grade glioma, medulloblastoma, ependymoma, DIPG and ATRT. Our hypothesis is that these patient-specific PDOX tumors will respond to anti-cancer therapies similarly to the corresponding human primary tumors, and the effective agents identified through this system would have better chances of clinical success. To test this hypothesis, we will perform a series of in vitro and in vivo assays to achieve the following aims: 1) to identify genetically accurate candidate PDOX models that bear the therapeutic target(s) of new investigational drugs through data mining of our mouse model molecular characterization databases; 2) to select the most responsive models through functional in vitro screening to determine time- and dose-responses; 3) to demonstrate therapeutic efficacy of new investigational drugs in multiple target-bearing PDOX models; and 4) to perform detailed analysis of cellular and molecular mechanisms of cell killing as well as the causes of therapy resistance both in vitro and in vivo. Our novel panel of PDOX mouse models represents a broad spectrum of genetic abnormalities of pediatric CNS tumors. All the assays are well established and routinely performed in our laboratory; we are uniquely positioned to accomplish the proposed drug studies in vivo. Our findings should provide strong preclinical evidence to support the initiation of clinical trials.

此申请是为响应资助机会而准备的：NCI儿科活体测试计划 (U01)，RFA-CA-20-034，以续订我们现有的PPTC UO1赠款。具体地说，我们建议继续体内的 我们的患者来源异种原位移植用于中枢神经系统肿瘤的检测程序 (PDOX)型号。脑瘤是儿童癌症相关死亡的主要原因。其中一个挑战是 在临床药物开发中，如何有效地对候选药物进行优先排序，以确保临床成功 癌症患者。然而，为此寻找新的抗癌药物的努力最有可能是有效的 由于缺乏具有临床相关性和分子水平的基因，临床应用已被封锁多年 精确的模型系统。幸运的是，我们已经建立了一个由150个儿童大脑PDOX模型组成的小组 通过将患者肿瘤标本直接注射到SCID小鼠的脑内来治疗肿瘤。这些PDOX型号 被证明复制了原始患者的组织病理学和主要基因异常 甚至在小鼠脑内进行连续亚移植期间也是如此。它们不仅代表着不同的 临床阶段(即在诊断、复发和晚期/尸检时)，但也复制了广泛的新的 确定了几乎所有类型的儿童脑肿瘤的分子亚型。异种移植瘤细胞也可以 为持续和按需供应致瘤的PDOX细胞而进行冷冻保存。此容量加在一起 通过我们优化的手术程序，我们每天可以植入多达260只小鼠，使 美国每年为每种肿瘤类型测试多种(例如，6-10种)药物。因此，我们的目标是利用 这一独特的PDOX模型小组用于检查新药物的治疗效果并分析机制 在高级别胶质瘤、髓母细胞瘤、室管膜瘤、DIPG和ATRT中的作用和治疗耐药。我们的 假设这些患者特异性的PDOX肿瘤对抗癌治疗的反应类似于 相应的人类原发肿瘤，通过这个系统确定的有效药物将具有 临床成功的机会更大。为了验证这一假设，我们将在体外和体内进行一系列 检测以实现以下目标：1)确定具有遗传准确性的候选PDOX模型 小鼠模型分子数据挖掘的新药治疗靶点(S) 特征数据库；2)通过功能体外筛选选择反应最灵敏的模型 确定时间和剂量反应；3)展示新研究药物的治疗效果 在多个靶向PDOX模型中；以及4)对细胞和分子进行详细分析 细胞杀伤的机制以及体内和体外治疗耐药的原因。我们的小说 PDOX小鼠模型小组代表了儿童中枢神经系统广泛的遗传异常 肿瘤。所有的化验都是在我们的实验室建立的，并且是常规的；我们是独一无二的 能够在体内完成拟议的药物研究。我们的发现应该会为临床前提供强有力的 支持启动临床试验的证据。