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: 10302832
• 负责人: Xiaonan Li
• 金额: $ 38.79万
• 依托单位: LURIE CHILDREN'S HOSPITAL OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10302832
• 关键词: Biological Assay; Biological Models; Brain; Brain Neoplasms; Cancer Etiology; Cancer Patient; Cells; Central Nervous System Neoplasms; Childhood; Childhood Brain Neoplasm; Childhood Central Nervous System Neoplasm; Clinic; Clinical; Clinical Drug Development; Clinical Trials; Cryopreservation; DNA copy number; Databases; Diffuse intrinsic pontine glioma; Dose; Ensure; Ependymoma; Funding; Gene Expression; Gene Mutation; Glioblastoma; Glioma; Human; Implant; In Vitro; Injections; Investigation; Investigational Drugs; Laboratories; Modeling; Molecular; Molecular Abnormality; Molecular Target; Mus; New Agents; Operative Surgical Procedures; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Positioning Attribute; Preclinical Testing; Primary Neoplasm; Research; Research Proposals; Resistance; SCID Mice; Series; Serum-Free Culture Media; Specimen; System; Testing; Time; Transplantation; Treatment Efficacy; Ursidae Family; Xenograft Model; Xenograft procedure; base; cancer therapy; cell killing; childhood cancer mortality; clinically relevant; data mining; drug candidate; drug testing; in vivo; in vivo evaluation; medulloblastoma; molecular modeling; mouse model; neoplastic cell; novel; novel therapeutics; pre-clinical; programs; response; screening; success; therapeutic target; tumor; tumor xenograft; tumorigenic

PROJECT SUMMARY/ABSTRACT This application is prepared for an extension of our previously funded research proposal that was in response to RFA-CA-14-018, Pediatric Preclinical Testing Consortium Research Programs (U01), and specifically to Type B: research Program for tumors of central nervous system (CNS) in vivo testing. 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. As drug candidates are more numerous than the small number of patients, it is essential to perform comprehensive pre-clinical testing to identify the investigational agents that are most likely to be effective in the clinic. However, such effort has been blocked for many years due to the lack of clinically relevant and molecularly accurate model systems. Fortunately, we have established a large panel (>130) of patient derived orthotopic xenograft (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 histopathological features, invasive phenotypes and major genetic abnormalities (gene expression, DNA copy number and gene mutations) of the original primary tumors even during serial sub-transplantations in vivo in mouse brains. 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 and ependymoma. 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.

项目概要/摘要 该申请是为了扩展我们之前资助的研究提案而准备的，该提案是为了响应 RFA-CA-14-018，儿科临床前测试联盟研究计划 (U01)，特别是 B型：中枢神经系统（CNS）肿瘤体内测试研究计划。脑肿瘤是 儿童癌症相关死亡的主要原因。临床药物开发的挑战之一是如何 有效地优先考虑候选药物，以确保癌症患者的临床成功。由于候选药物越来越多 由于患者数量多于少数，因此有必要进行全面的临床前测试 确定最有可能在临床中有效的研究药物。然而，这样的努力已经 由于缺乏临床相关和分子精确的模型系统，多年来一直受到阻碍。 幸运的是，我们已经建立了一个大型面板（>130）患者来源的原位异种移植（PDOX）模型 通过将患者肿瘤标本直接注射到 SCID 小鼠的大脑中来治疗小儿脑肿瘤。 这些 PDOX 模型被证明复制了组织病理学特征、侵袭性表型和 原始原代的主要遗传异常（基因表达、DNA 拷贝数和基因突变） 甚至在小鼠大脑体内的连续亚移植过程中也能抑制肿瘤的发生。异种移植肿瘤细胞也可以 冷冻保存以持续按需供应致瘤性 PDOX 细胞。此容量与 我们优化的手术程序每天可以植入多达 260 只小鼠，这使我们成为可能 每年针对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点 独特的 PDOX 模型组，用于检查新药的治疗效果并分析其机制 高级别神经胶质瘤、髓母细胞瘤和室管膜瘤的作用和治疗耐药性。我们的假设是 这些患者特异性 PDOX 肿瘤对抗癌疗法的反应与相应的人类相似 原发性肿瘤，通过该系统识别出的有效药物将有更好的临床机会 成功。为了检验这一假设，我们将进行一系列体外和体内测定，以实现以下目标 目标：1) 鉴定具有新治疗靶点的遗传准确的候选 PDOX 模型 通过对我们的小鼠模型分子表征数据库进行数据挖掘来研究药物； 2）到 通过功能性体外筛选选择反应最灵敏的模型，以确定时间和剂量 回应； 3) 证明新研究药物对多靶点PDOX的治疗效果 模型； 4) 详细分析细胞杀伤的细胞和分子机制以及 体外和体内治疗耐药的原因。我们新颖的 PDOX 小鼠模型组代表了 儿童中枢神经系统肿瘤的广泛遗传异常。所有检测均已建立并 在我们的实验室常规进行；我们处于独特的地位来完成拟议的药物研究 体内。我们的研究结果应提供强有力的临床前证据来支持临床试验的启动。