Genetic Mouse Models of Glioma

神经胶质瘤的遗传小鼠模型

• 批准号: 8839207
• 负责人: Luis Fernando Parada
• 金额: $ 17.92万
• 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 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8839207
• 关键词: Ablation; Accounting; Adult; Brain; Brain Neoplasms; Breeding; CSPG4 gene; Cells; Characteristics; Clinical; Data; Development; Developmental Biology; Diphtheria Toxin; Dorsal; Event; Funding; Ganciclovir; Genes; Genetic; Genetically Engineered Mouse; Glial Fibrillary Acidic Protein; Glioblastoma; Glioma; Gliomagenesis; Grant; Health; Human; Incidence; Induced Mutation; Investigation; Label; Malignant Glioma; Malignant Neoplasms; Malignant neoplasm of brain; Masks; Methods; Modeling; Molecular; Morphologic artifacts; Mus; Mutate; Mutation; NF1 gene; NF1 tumor suppressor; Natural History; Nature; Neoplasm Metastasis; Oligodendroglia; PDGFRB gene; Pathologist; Patients; Penetrance; Physiological; Population; Property; Recurrence; Research; Resistance; Resort; Role; Sampling; Somatic Mutation; Source; Staging; Stem cells; Surveys; System; Tamoxifen; Techniques; Testing; The Cancer Genome Atlas; Therapeutic; Thymidine Kinase; Transgenes; Transgenic Organisms; Transplantation; Tumor Suppressor Genes; Tumor Suppressor Proteins; Up-Regulation; base; brain cell; cancer stem cell; cell suicide; chemotherapy; design and construction; diphtheria toxin receptor; functional genomics; human DICER1 protein; improved; in vivo; insight; medulloblastoma; mouse model; mutant; neoplastic cell; nerve stem cell; nestin protein; novel; novel therapeutics; outcome forecast; progenitor; recombinase; research study; response; stem; stem cell niche; stem cell population; success; temozolomide; tool; transcriptome sequencing; transgene expression; tumor; tumor initiation; tumorigenesis; tumorigenic

DESCRIPTION (provided by applicant): Glioblastoma multiforme (GBM) is an aggressive and incurable cancer. The dire prognosis for this most prevalent form of malignant brain tumor is unchanged over 30 years. The Cancer Genome Atlas (TCGA) project has genomically surveyed several hundred human GBM samples and identified a small spectrum of genes that are frequently somatically mutated and include the tumor suppressors NF1, p53 and Pten. We have generated and studied mouse models that spontaneously form classic GBM with 100% incidence. The models are based on somatic mutation of NF1, p53, and Pten. The full penetrance of these tumors has allowed us to probe early pre-tumorigenic events and to identify the adult neural stem cell (NSC) niche as the prevalent source of these tumors. In the previous funding period, we exploited these models to demonstrate a stem cell origin of these tumors and exclude differentiated brain cells as a source. We probed transcriptional profiles to identify glioma-specific upregulation of HoxA genes and examined their function. We developed tools to directly test and demonstrate in vivo the existence of a hierarchical population of cancer stem cells (CSCs) by direct lineage tracing in spontaneous gliomas without resorting to ex vivo transplantation and/or culturing. We unveiled a novel GBM that arises from a different cell of origin. Finally, we developed a novel model of metastatic medulloblastoma that depends on Dicer mutation and is being developed as a separate research initiative. The present proposal embodies two Specific Aims. Aim 1, entitled: "Cancer stem cells in glioma: identification, isolation and characterization." pursues our recent success in transgenically labeling an endogenous glioma cell population that is responsible for tumor recurrence after chemotherapy. We have devised an advanced NSC- and CSC-specific transgene that will co-express Cre recombinase, eGFP, and human diphtheria toxin receptor. This transgene, when bred into the Nf1;p53;Pten floxed background, will permit more precise identification and study of the relatively quiescent CSC population. It will allow purification for quantitative RNA seq analysis i comparison to the non-CSC tumor population, and to the progenitor NSC population. These studies will provide novel information and insight into these newly discovered CSCs. Aim 2 will examine whether "neural progenitor cells (NPCs) and oligodendrocyte progenitor cells (OPCs) give rise to distinct forms of glioma and GBM". We provide evidence that NPCs and OPCs can, by mutations in NF1, p53 and Pten, give rise to gliomas. Mutant NPCs give rise to tumors very similar to the tumors arising in the stem cell compartments whereas OPCs generate an independent form of GBM with distinct molecular, and developmental characteristics. We propose to study these novel tumors in depth since, to the pathologist, they belong to a single GBM pool. Our detailed studies of these tumors may inform human tumor analysis and serve to separate these OPC GBMs that may have different properties, including response to therapies. Genomic, functional, and CSC studies will be pursued for these novel brain tumors.

描述（由申请人提供）：多形胶质母细胞瘤（GBM）是一种侵略性且无法治愈的癌症。这种最普遍的恶性脑肿瘤的可怕预后在30年内没有改变。癌症基因组图集（TCGA）项目已经对数百个人类GBM样品进行了研究，并确定了一小部分基因，这些基因经常被体形突变，并包括肿瘤抑制剂NF1，p53和PTEN。我们已经生成和研究了小鼠模型，这些模型自发地形成了经典的GBM，其发病率为100％。这些模型基于NF1，p53和PTEN的体细胞突变。这些肿瘤的全部渗透性使我们能够探测早期的肿瘤前事件，并确定成年神经干细胞（NSC）壁ni，成为这些肿瘤的普遍来源。在上一个资金期间，我们利用这些模型来证明这些肿瘤的干细胞起源，并将分化的脑细胞作为来源。我们探测了转录谱以鉴定HOXA基因的胶质瘤特异性上调并检查其功能。我们开发了直接测试和证明体内的工具，通过直接在自发的神经胶质瘤中直接谱系跟踪而无需诉诸于体内移植和/或培养，从而存在癌症干细胞（CSC）的分层群体。我们推出了一种新型的GBM，该GBM由不同的原始细胞产生。最后，我们开发了一种新型的转移性髓母细胞瘤模型，该模型依赖于DICER突变，并正在作为单独的研究计划开发。本提案体现了两个具体目标。 AIM 1，题为：“神经胶质瘤中的癌症干细胞：鉴定，分离和表征”。追求我们最近在转基因标记内源性神经胶质瘤细胞种群方面取得成功，该细胞群是导致化疗后肿瘤复发的。我们设计了一个晚期的NSC和CSC特异性转基因，该转基因将共表达CRE重组酶，EGFP和人二胺毒素受体。当繁殖到NF1; p53; pten的背景中时，这种转基因将允许对相对静止的CSC种群进行更精确的识别和研究。它将允许纯化用于定量RNA SEQ分析I与非CSC肿瘤种群以及祖细胞NSC种群进行比较。这些研究将为这些新发现的CSC提供新颖的信息和洞察力。 AIM 2将检查“神经祖细胞（NPC）和少突胶质细胞祖细胞（OPC）是否会产生不同形式的神经胶质瘤和GBM”。我们提供的证据表明，NPC和OPC可以通过NF1，p53和PTEN中的突变产生神经胶质瘤。突变的NPC产生与干细胞区室中产生的肿瘤非常相似的肿瘤，而OPC产生了一种独立的GBM形式，具有独特的分子和发育特征。我们建议对这些新型肿瘤进行深入研究，因为他们对病理学家属于一个GBM池。我们对这些肿瘤的详细研究可能会为人类肿瘤分析提供信息，并将可能具有不同特性的OPC GBM分开，包括对疗法的反应。这些新型脑肿瘤将进行基因组，功能和CSC研究。