Genetic Mouse Models of Glioma

神经胶质瘤的遗传小鼠模型

• 批准号: 8010613
• 负责人: Luis Fernando Parada
• 金额: $ 39.03万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8010613
• 关键词: Ablation; Address; Adult; Alleles; Anaplastic astrocytoma; Appearance; Astrocytoma; Attention; Brain Neoplasms; Brain region; Cell surface; Cells; Central Nervous System Neoplasms; Complement; Coupled; Data; Development; Diagnostic Neoplasm Staging; Differentiation and Growth; Drug Delivery Systems; Event; Gene Expression; Genes; Genetic; Genomics; Genotype; Glioblastoma; Glioma; Growth; Health; Histologic; Human; Investigation; Knock-out; Laboratories; Loss of Heterozygosity; Malignant Neoplasms; Methods; MicroRNAs; Microarray Analysis; Modeling; Molecular; Molecular Profiling; Mouse Strains; Mus; Mutant Strains Mice; Mutate; Mutation; NF1 gene; NF1 tumor suppressor; Nature; Neoplasm Metastasis; Nervous system structure; Neural Crest; Neurofibromatoses; Neuroglia; Nude Mice; Oncogenic; PTEN gene; Pathway interactions; Patients; Penetrance; Penetrance analysis; Peripheral Nervous System Neoplasms; Physiological; Platelet-Derived Growth Factor Receptor; Process; Property; Protein p53; RNA Interference; Radial; Reporting; Research; Research Design; Risk; Role; Signal Pathway; Stem cells; Tamoxifen; Techniques; Testing; The Cancer Genome Atlas; Therapeutic; Thymidine Kinase; Tissues; Transgenic Mice; Transplantation; Tumor Suppressor Genes; Tumor Suppressor Proteins; Tumor Tissue; Tumor stage; Work; adult stem cell; base; cancer stem cell; cell type; clinically relevant; design; epidermal growth factor receptor VIII; follow-up; insight; interest; mouse model; mutant; neonate; neoplastic cell; nerve stem cell; nestin protein; neurodevelopment; neurofibroma; neuropathology; novel; outcome forecast; overexpression; progenitor; promoter; relating to nervous system; research study; stem; stem cell niche; stem cell population; subventricular zone; therapy development; tool; tumor; tumor initiation; tumor progression; tumorigenesis; tumorigenic

DESCRIPTION (provided by applicant): Malignant astrocytomas are brain tumors that are locally infiltrative and incurable, with poor prognosis for the patient. Despite profound therapeutic implications, the identity of the cell(s) of origin of these tumors has not been rigorously determined. In addition, glioblastoma multiforme, the most prevalent and deadly form of brain tumor, can either progress from lower grade astrocytic gliomas or arise de novo, the mechanisms by which are not well understood. We previously reported mouse models based on conditional inactivation of human astrocytoma-relevant tumor suppressors p53, Nf1, and Pten, wherein through somatic loss of heterozygosity, mutant mice develop tumors that histologically and molecularly resemble human astrocytomas with 100% penetrance. In the present application, we propose experiments designed to investigate the cell(s) of tumor origin and to extend our analyses of tumorigenesis in these mouse models. To this end, Specific Aim 1 rigorously tests our hypothesis that gliomas originate in stem/progenitor cells. Our research design utilizes genetic and stereotactic methods to mutate the tumor suppressor genes specifically in the stem cell niche. We will also use a genetic method to ablate these neural stem cells in our tumor models and then analyze the effect on tumor formation. Specific Aim 2 will exploit our ability to culture fresh tumor tissue as self-renewable neurospheres to further characterize our Nf1;p53;Pten "de novo" glioma mouse model. We will evaluate their growth and differentiation properties, as well as their tumorigenic potential via transplantation techniques. Additionally, using a stem cell-specific GFP transgenic mouse, we will analyze the expression of candidate cell surface markers with the aim of identifying "signature markers" that will allow us to prospectively isolate the cancer stem cells. Specific Aim 3 will employ microarray analyses to identify the gene expression profiles that correlate with de novo vs. progressive glioma, using neurospheres derived from Nf1;p53;Pten and Nf1;p53 tumors, respectively. We also propose to analyze tissue from pre-symptomatic mice in order to gain insight into the early molecular events of tumor initiation. Potentially interesting genes will be functionally pursued using RNAi and overexpression techniques. As microRNAs have recently been implicated in glioma, we will also use microarray analysis to identify microRNAs that are differentially regulated in our tumor models and also investigate the role of candidate microRNAs in glioma. Our fully penetrant glioma mouse models are clinically relevant and powerful tools for identifying and functionally characterizing novel genes and pathways that may be therapeutically tractable in human glioma. PUBLIC HEALTH RELEVANCE: Malignant astrocytomas are the most common type of brain tumors that occur in adults and due to their infiltrative and aggressive nature, are virtually incurable. The focus of our research is to use mouse brain tumor models to identify and analyze genes that are involved in brain tumor initiation and progression. The hope is that by identifying these genes, we might begin to find suitable drug targets and develop therapies that would eliminate or inhibit the growth of these devastating tumors.

描述（申请人提供）：恶性星形细胞瘤是一种局部浸润性脑肿瘤，无法治愈，患者预后较差。尽管具有深远的治疗意义，但这些肿瘤起源的细胞的身份尚未得到严格确定。此外，多形性胶质母细胞瘤是最常见和致命的脑肿瘤形式，它可以从低级别星形细胞胶质瘤进展，也可以从头出现，其机制尚不清楚。我们之前报道过基于人星形细胞瘤相关肿瘤抑制因子 p53、Nf1 和 Pten 的条件失活的小鼠模型，其中通过杂合性的体细胞丧失，突变小鼠产生在组织学和分子上类似于人星形细胞瘤的肿瘤，外显率为 100%。在本申请中，我们提出了旨在研究肿瘤起源细胞并扩展我们对这些小鼠模型中肿瘤发生的分析的实验。为此，特定目标 1 严格检验了我们的假设，即神经胶质瘤起源于干/祖细胞。我们的研究设计利用遗传和立体定向方法来突变干细胞生态位中的肿瘤抑制基因。我们还将使用遗传方法消除肿瘤模型中的这些神经干细胞，然后分析对肿瘤形成的影响。具体目标 2 将利用我们培养新鲜肿瘤组织作为自我更新神经球的能力，以进一步表征我们的 Nf1;p53;Pten“从头”神经胶质瘤小鼠模型。我们将评估它们的生长和分化特性，以及通过移植技术的致瘤潜力。此外，使用干细胞特异性 GFP 转基因小鼠，我们将分析候选细胞表面标记的表达，目的是识别“特征标记”，使我们能够前瞻性地分离癌症干细胞。具体目标 3 将分别使用源自 Nf1;p53;Pten 和 Nf1;p53 肿瘤的神经球，采用微阵列分析来识别与新生和进展性神经胶质瘤相关的基因表达谱。我们还建议分析症状前小鼠的组织，以深入了解肿瘤发生的早期分子事件。将使用 RNAi 和过表达技术对潜在感兴趣的基因进行功能性研究。由于 microRNA 最近与神经胶质瘤有关，我们还将使用微阵列分析来识别在我们的肿瘤模型中受到差异调节的 microRNA，并研究候选 microRNA 在神经胶质瘤中的作用。我们的完全渗透性神经胶质瘤小鼠模型是临床相关且强大的工具，用于识别和功能表征人类神经胶质瘤中可能易于治疗的新基因和通路。公共卫生相关性：恶性星形细胞瘤是成人中最常见的脑肿瘤类型，由于其浸润性和侵袭性，实际上是无法治愈的。我们研究的重点是使用小鼠脑肿瘤模型来识别和分析参与脑肿瘤发生和进展的基因。希望通过识别这些基因，我们可以开始寻找合适的药物靶点并开发能够消除或抑制这些破坏性肿瘤生长的疗法。