Characterizing the glioma cell of origin in vivo using MADM, a mouse genetic mosa

使用 MADM（一种小鼠遗传莫萨）表征体内神经胶质瘤细胞的起源

基本信息

批准号：
8252212
负责人：
Hui Zong
金额：
$ 27.45万
依托单位：
UNIVERSITY OF OREGON
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2012-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8252212
关键词：
Address Animal Model Cell Lineage Cell division Cells Characteristics Development Developmental Process Disease Genes Genetic Glioma Gliomagenesis Grant Green Fluorescent Proteins Human Label Laboratories Lead Malignant - descriptor Malignant Neoplasms Malignant neoplasm of brain Medicine Modeling Mus Mutate Mutation Neurofibromatosis 1 Pathway interactions Pharmaceutical Preparations Population Premalignant Property Recruitment Activity Reporting Research Resolution Specificity Staging Stem cells Suppressor Mutations System Testing Therapeutic Therapeutic Intervention Tissues Transgenic Mice Transplantation Tumor Stem Cells Tumor Suppressor Genes Tumor Suppressor Proteins Work base cancer stem cell cell type design effective therapy in vivo mouse model mutant neoplastic cell nerve stem cell novel oligodendrocyte precursor prevent progenitor public health relevance stemness therapeutic target treatment strategy tumor tumor initiation tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Glioma is the most common type of malignant brain tumor. Despite widespread advances in cancer medicine, this devastating disease remains incurable. Recently, it was reported that gliomas contain a population of tumor stem cells that can form self renewable tumor spheres in culture and re- initiate gliomas after transplantation into immuno-suppressed mice. Targeting these tumor stem cells is an exciting prospect towards a glioma cure. However, critical information required to develop such therapeutic strategies, including the developmental origin of the glioma stem cells, remains unknown. To address the tumor cell of origin problem, it requires the use of glioma animal models that allow the analysis of early-stage pre-malignant tumor cells. Unfortunately, current mouse models cannot provide adequate in vivo resolution for such studies. Our laboratory has developed a new mouse glioma model based on a novel genetic mosaic system termed MADM (Mosaic Analysis with Double Markers, Zong et al Cell 2005). Using MADM, we can generate rare, green fluorescent protein (GFP)-labeled neural stem cells (NSCs) that are double null for two key tumor suppressor genes, p53 and Neurofibromatosis Type 1 (NF1), within an otherwise normal mouse. This approach allows us to analyze the entire course of gliomagenesis with single-cell resolution in vivo. Our preliminary findings show that, although the mutations are generated specifically in NSCs, resulting glioma cells manifest many cellular features of oligodendrocyte precursors (OPCs). Prior to malignancy, OPCs are the only cell lineage that drastically over-expands in the MADM mice. In the glioma tumor mass, OPCs are also the predominant cell type that maintains active cell divisions. When we purify these OPC-like glioma cells they manifest salient glioma stem cell features, including forming renewable tumor spheres, differentiating into multiple cell lineages, and reinitiating gliomas after being transplanted into immuno-suppressed mice. Based on these preliminary results, we will test the following hypothesis: 1) OPCs are the key cell type that initiates and renews gliomagenesis; 2) mutant OPCs can de-differentiate to acquire stem cell properties; and 3) targeting OPCs or their stem-cell characteristics will be effective treatment strategies for gliomas. Our proposed studies will lead to valuable basic understanding of the developmental process of gliomas. The identification of tumor-initiating cells should provide a basis for designing rationale treatment strategies for the cure. Conceptually, our proposed work explores the uncharted territory of tumor initiation, and provides critical groundwork for the refinement of mouse models for mechanistic understanding of human cancers. PUBLIC HEALTH RELEVANCE: Studies proposed in this grant aim at the identification of the origin of cancer stem cells for glioma, a currently incurable disease. A thorough understanding of these cells should enable the design of rationale therapeutic strategies by targeting specific cells or genes to treat gliomas. As a consequence, the enhanced specificity should lead to more effective and less toxic drugs, which could eventually provide a cure for gliomas.

描述（由申请人提供）：神经胶质瘤是最常见的恶性脑肿瘤类型。尽管癌症医学方面取得了广泛的进步，但这种毁灭性疾病仍然无法治愈。最近，据报道，神经胶质瘤含有肿瘤干细胞的群体，可以在培养物中形成可再生的肿瘤球，并在移植到免疫抑制的小鼠中后重新构成神经胶质瘤。靶向这些肿瘤干细胞是对神经胶质瘤治疗的令人兴奋的前景。但是，开发此类治疗策略所需的关键信息，包括神经胶质瘤干细胞的发育起源，仍然未知。为了解决起源问题的肿瘤细胞，它需要使用胶质瘤动物模型，以分析早期恶性肿瘤细胞。不幸的是，当前的小鼠模型无法为此类研究提供足够的体内分辨率。我们的实验室基于一种称为MADM的新型遗传镶嵌系统开发了一种新的小鼠胶质瘤模型（带有双标记的镶嵌分析，Zong等人Cell 2005）。使用MADM，我们可以生成稀有的，绿色的荧光蛋白（GFP）标记的神经干细胞（NSC），对于两种抑制的两个关键肿瘤抑制基因，p53和神经纤维瘤病1型（NF1），它们在原本正常的小鼠中为双重null。这种方法使我们能够通过体内单细胞分辨率分析整个神经胶质作用的过程。我们的初步发现表明，尽管突变是在NSC中特别生成的，但胶质瘤细胞表现出少突胶质细胞前体（OPC）的许多细胞特征。在恶性肿瘤之前，OPC是唯一在MADM小鼠中急剧过度膨胀的细胞谱系。在神经胶质瘤肿瘤质量中，OPC也是维持活性细胞分裂的主要细胞类型。当我们纯化这些类似OPC的神经胶质瘤细胞时，它们表现出显着的神经胶质瘤干细胞特征，包括形成可再生肿瘤球，分化为多个细胞谱系，并在被移植到免疫抑制的小鼠中后重新启动神经胶质瘤。基于这些初步结果，我们将测试以下假设：1）OPC是启动和更新神经胶质作用的关键细胞类型； 2）突变型OPC可以取消分化以获取干细胞特性； 3）针对OPC或其干细胞特征将是神经胶质瘤的有效治疗策略。我们提出的研究将导致对神经胶质瘤发展过程的宝贵基本理解。识别肿瘤发射细胞应为设计治疗的理由治疗策略提供基础。从概念上讲，我们提出的工作探讨了肿瘤启动的未知领域，并为小鼠模型的完善提供了关键的基础，以便对人类癌症的机械理解。公共卫生相关性：该赠款中提出的研究旨在鉴定癌症干细胞的起源（一种目前无法治愈的疾病）。对这些细胞的透彻理解应通过靶向特定细胞或基因来治疗神经胶质瘤来实现理由治疗策略的设计。结果，增强的特异性应导致更有效和毒性更少的药物，这最终可以治愈神经胶质瘤。