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

基本信息

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
项目类别：
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

项目摘要

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.

该申请是根据资金机会来准备的：NCI小儿体内测试计划（U01），RFA-CA-20-034，以更新我们现有的PPTC UO1赠款。具体来说，我们建议继续体内中枢神经系统（CNS）肿瘤的测试程序使用我们的患者衍生的原位异种移植（PDOX）模型。脑肿瘤是儿童与癌症相关死亡的主要原因。挑战之一在临床药物开发中，如何有效优先考虑候选药物以确保临床成功癌症患者。但是，确定新的反癌代理商的努力很可能是有效的由于缺乏临床相关和分子，在诊所中已被阻塞多年准确的模型系统。幸运的是，我们已经建立了一个150个PDOX小儿大脑模型的面板通过直接注射患者肿瘤标本进入SCID小鼠的大脑中的肿瘤。这些PDOX模型被证明已经复制了原始患者的组织病理学和主要遗传异常即使在小鼠大脑体内串行亚移植期间，肿瘤也是如此。他们不仅代表不同的临床阶段（即在诊断，复发和末端/尸检时），但也复制了新的鉴定出几乎所有类型的小儿脑肿瘤的分子亚型。异种移植肿瘤细胞也可以冷冻保存，以持续和按需供应肿瘤性PDOX细胞的供应。这种能力总和通过我们优化的外科手术，我们每天最多可以植入260只鼠标，使其成为可能我们每年对每年的多种药物（例如6-10）进行测试。因此，我们的目标是利用这种独特的PDOX模型，以检查新药物的治疗功效并分析机制高级胶质瘤，髓母细胞瘤，室心瘤，DIPG和ATRT的作用和治疗耐药性。我们的假设是这些患者特异性的PDOX肿瘤将对抗癌疗法有反应相应的人类原发性肿瘤以及通过该系统鉴定的有效药物将具有更好的临床成功机会。为了检验这一假设，我们将执行一系列体外和体内实现以下目的的测定：1）确定遗传准确的候选PDOX模型通过我们的小鼠模型分子的数据挖掘的新研究药物的治疗靶标表征数据库； 2）通过在体外筛选中选择最响应的模型确定时间和剂量回答； 3）证明新研究药物的治疗功效在多个携带目标的PDOX模型中； 4）进行细胞和分子的详细分析细胞杀伤的机制以及体外和体内耐药性的原因。我们的小说 PDOX小鼠模型的面板代表了小儿中枢神经系统的一系列遗传异常肿瘤。所有测定法在我们的实验室中都得到了很好的确定和常规性；我们是独特的定位在体内完成拟议的药物研究。我们的发现应该提供强大的临床前支持临床试验开始的证据。