An Investigation of Radial Glia as the Source of Ependymoma Stem Cells

描述(申请人提供)：室管膜瘤是脑和脊髓的肿瘤。这种疾病的治疗方法和死亡率在过去20年中几乎没有变化，这突显了对新疗法的巨大需求。室管膜瘤在组织学上的相似性导致研究人员将这些肿瘤作为一个单一实体来对待；但我们已经证明，来自中枢神经系统不同区域的室管膜瘤包括不同的临床和分子亚型，这表明它们是不同的疾病。因此，当代治愈所有室管膜瘤患者的努力必须关注于了解这些疾病亚型的生物学基础，并在必要时开发针对亚型的治疗方法。在上一个资助周期中，我们开发了一种跨物种基因组学方法，描述了人类室管膜瘤的基因组亚型，并将这些亚型与小鼠的神经干细胞进行匹配，以产生准确的疾病模型。我们将在这项工作的基础上完成三个具体目标，旨在测试核心假设，即室管膜瘤亚型是由不同的细胞信号驱动的，这些信号可以靶向获得治疗收益。目标1将 使用新的‘病毒库’筛查来测试前90个候选癌基因和40个肿瘤抑制基因在体内的致癌作用，这些基因是我们之前在室管膜瘤中发现的。因此，我们将产生一种概括了人类室管膜瘤亚型全谱的小鼠“临床”。AIM 2将使用高通量药物筛选、全基因组结合和细胞生物学分析来询问在AIM 1中开发的模型，以确定维持每种室管膜瘤亚型的关键细胞信号，从而可能成为治疗靶点。将这些数据与人类和小鼠肿瘤的基因分析相结合，将进一步验证候选药物靶点。以AIM 2中确定的信号为靶点的治疗的未来临床试验 将要求选择具有适当疾病亚型的患者。因此，Aim 3将开发一种强大的Affymetrix定量基因检测方法，使用福尔马林固定的石蜡包埋组织可靠而快速地诊断室管膜瘤亚型。 与公共卫生的相关性：室管膜瘤患者的治疗未能取得进展是由于缺乏临床前模型和药物开发过程，该过程没有考虑到构成这种疾病的离散亚型。我们的计划将开发高精度的室管膜瘤全谱小鼠模型，并使用这些模型来识别强大的药物靶点和诊断工具，用于特定亚型的临床试验。

DESCRIPTION (provided by applicant): Ependymomas are tumors of the brain and spinal cord. Treatment approaches and mortality rates for this disease have changed little over the last twenty years, highlighting the great need for new therapies. Histologic similarities among ependymomas have led investigators to treat these tumors as a single entity; but we have shown that ependymomas from different regions of the central nervous system include discrete clinical and molecular subtypes, suggesting they are different diseases. Thus, contemporary efforts to cure all patients with ependymoma must be concerned with understanding the biological basis of these disease subtypes, and where necessary, developing subtype-specific therapies. During the last funding cycle we developed a cross- species genomics approach that characterized genomic subtypes of human ependymoma and matched these with neural stem cells in the mouse to generate accurate models of the disease. We will build on this work to complete three Specific Aims designed to test the central hypothesis that 'Ependymoma subtypes are driven by distinct cell signals that can be targeted for therapeutic gain.' Aim 1 will employ new 'virus-pool' screens to test the in vivo oncogenic role of the top 90 candidate oncogenes and 40 tumor suppressor genes that we previously identified in ependymoma. We will thereby generate a 'clinic' of mice that recapitulate the full spectrum on human ependymoma subtypes. Aim 2 will interrogate the models developed in Aim 1 using high- throughput drug screens, kinome-wide binding and cell biology assays to pinpoint the key cell signals that maintain each ependymoma subtype that might therefore serve as therapeutic targets. Integration of these data with genetic analyses of human and mouse tumors will further validate candidate drug targets. Future clinical trials of therapies that target signals identified in Aim 2 will demand the selection of patients with the appropriate disease subtype. Therefore, Aim 3 will develop a robust Affymetrix Quantigene assay to reliably and rapidly diagnose ependymoma subtypes using formalin fixed paraffin embedded tissue. PUBLIC HEALTH RELEVANCE: The failure to progress the treatment of patients with ependymoma has resulted from a lack of preclinical models and a drug development process that does not account for the discrete subtypes that comprise the disease. Our proposal will develop highly accurate mouse models of the full spectrum of ependymoma and use these to identify robust drug targets and diagnostic tools for subtype-specific clinical trials.