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.

项目总结/摘要 此应用程序是为我们以前资助的研究计划的扩展而准备的， RFA-CA-14-018，儿科临床前试验联盟研究项目（U 01），特别是 B类：中枢神经系统（CNS）肿瘤体内试验研究项目。脑瘤是 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。随着候选药物越来越多 由于患者人数众多，因此必须进行全面的临床前检测， 确定最有可能在临床上有效的研究药物。然而，这种努力 由于缺乏临床相关的和分子精确的模型系统，多年来一直被封锁。 幸运的是，我们已经建立了一个大的面板（>130）的患者来源的原位异种移植（PDOX）模型 通过将患者的肿瘤标本直接注射到SCID小鼠的脑中， 这些PDOX模型显示具有复制的组织病理学特征、侵袭性表型和 原始原发灶的主要遗传异常（基因表达、DNA拷贝数和基因突变） 甚至在小鼠脑中的体内连续亚移植期间也是如此。异种移植肿瘤细胞也可以是 冷冻保存用于致瘤PDOX细胞的持续和按需供应。这种能力与 我们优化的手术程序，我们每天可以植入多达260只老鼠，使我们有可能 为了测试多个（例如，6-10）药物每年每种肿瘤类型。因此，我们的目标是利用这一点， 独特的PDOX模型面板，用于检查新药物的治疗效果并分析 在高级别胶质瘤、髓母细胞瘤和室管膜瘤中的作用和耐药性。我们的假设是 这些患者特异性PDOX肿瘤将与相应的人PDOX肿瘤类似地对抗癌疗法作出反应。 原发性肿瘤，通过该系统鉴定的有效药物将有更好的临床应用机会。 成功为了验证这一假设，我们将进行一系列体外和体内试验，以实现以下目标 目的：1）鉴定携带新的治疗靶标的遗传上准确的候选PDOX模型， 通过我们的小鼠模型分子表征数据库的数据挖掘研究药物; 2） 通过功能性体外筛选选择最有效的模型，以确定时间和剂量， 缓解; 3）证明新研究药物在多靶点PDOX中的疗效 模型;和4）进行细胞杀伤的细胞和分子机制的详细分析，以及 在体外和体内的治疗抗性的原因。我们的新型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。所有检测方法均已确立， 常规在我们的实验室进行;我们处于独特的地位，以完成拟议的药物研究， vivo.我们的研究结果应提供强有力的临床前证据，以支持临床试验的启动。