SOMATIC CELL TRANSFER TO MODEL MEDULLOBLASTOMA IN MICE

体细胞转移至小鼠髓母细胞瘤模型

• 批准号: 7243512
• 负责人: DANIEL WEBSTER FULTS
• 金额: $ 21.33万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7243512
• 关键词: 1-Phosphatidylinositol 3-Kinase; Abbreviations; Antineoplastic Agents; Apoptosis; Automobile Driving; Avian Leukosis Virus; Biology; Brain; Cell Line; Cell Proliferation; Cell Surface Receptors; Cells; Cerebellum; Child; Clinical; Cultured Cells; Cytoplasmic Granules; Development; Drosophila genus; Erinaceidae; Gene Combinations; Gene Expression; Gene Transfer; Genes; Glial Differentiation; Growth; Homologous Gene; IGF1 gene; IGF2 gene; In Vitro; Individual; Insulin-Like Growth Factor I; Insulin-Like Growth Factor II; Integral Membrane Protein; Intermediate Filament Proteins; Malignant neoplasm of brain; Mediating; Modeling; Molecular; Mus; Neurons; Newborn Infant; Notch Signaling Pathway; Oncogene Proteins; Oncogenes; Pathway interactions; Phosphatidylinositols; Phosphoinositide-3-Kinase, Catalytic, Gamma Polypeptide; Phosphorylation; Preclinical Testing; Primitive Neuroectodermal Tumor; Proteins; Relative (related person); Research; Research Personnel; Retroviral Vector; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Somatic Cell; Somatomedins; Stem cells; Subgroup; System; Testing; Transgenes; Transgenic Mice; cell growth; chemotherapeutic agent; established cell line; human SMO protein; immunoreactivity; in vivo; inhibitor/antagonist; insight; insulin receptor substrate 1 protein; medulloblastoma; morphogens; mouse model; neoplastic cell; nerve stem cell; nestin protein; neurodevelopment; notch protein; progenitor; programs; promoter; receptor; relating to nervous system; therapeutic target; tissue culture; tumor; tumor growth; vector

DESCRIPTION (provided by applicant): The clinical focus of this research is medulloblastoma (MB), a malignant brain tumor that arises in the cerebellum in children. The main limitation to current therapy is collateral damage to the developing brain by anti-cancer agents. Insights into new treatments that interfere with tumor cell growth without impeding neural development will come from an increased understanding of the basic biology of MB. The overall objective of this research is to use a genetically defined, mouse model of MB, which we created using the RCAS/tv-a system, to identify key signaling proteins in MB formation. This system utilizes a retroviral vector (RCAS), derived from avian leukosis virus, subgroup A (ALV-A), and a transgenic mouse line (Ntv-a) that expresses TVA (the receptor for ALV-A) under control of the nestin gene promoter. The nestin gene encodes an intermediate filament protein expressed by neural progenitor cells prior to their commitment to neuronal or glial differentiation. This makes it possible to target expression of exogenous genes specifically to nestin-expressing neural progenitors in the cerebellum - the cells-of-origin for MB. Specific aim 1 is to generate in vivo mouse models of MB by using the RCAS/tv-a system to transfer combinations of genes that stimulate proliferation, block differentiation, and inhibit apoptosis in neural progenitor cells. Specific aim 2 is to establish cell lines from MBs generated by RCAS-mediated gene transfer. Using these cell lines, we will pharmacologically block the individual signaling pathways activated by retroviral gene transfer in order to assess the relative contribution of proliferation, differentiation, and apoptosis to tumor growth. The results of this study will provide insights into signal transduction pathways that promote MB growth. This information can be used to identify molecular therapeutic targets and facilitate development of preclinical testing models for chemotherapeutic agents.

描述（由申请人提供）：本研究的临床重点是髓母细胞瘤（MB），这是一种发生在儿童小脑的恶性脑肿瘤。目前治疗的主要限制是抗癌药物对发育中的大脑造成附带损害。对干扰肿瘤细胞生长而不阻碍神经发育的新疗法的见解将来自于对 MB 基础生物学的加深了解。本研究的总体目标是使用基因定义的 MB 小鼠模型（我们使用 RCAS/tv-a 系统创建该模型）来识别 MB 形成中的关键信号蛋白。该系统利用源自禽白血病病毒 A 亚型 (ALV-A) 的逆转录病毒载体 (RCAS) 和在巢蛋白基因启动子控制下表达 TVA（ALV-A 受体）的转基因小鼠系 (Ntv-a)。巢蛋白基因编码神经祖细胞在神经元或神经胶质分化之前表达的中间丝蛋白。这使得将外源基因的表达特异性靶向小脑中表达巢蛋白的神经祖细胞（MB 的起源细胞）成为可能。具体目标 1 是通过使用 RCAS/tv-a 系统转移刺激神经祖细胞增殖、阻止分化和抑制凋亡的基因组合来生成 MB 体内小鼠模型。具体目标 2 是利用 RCAS 介导的基因转移产生的 MB 建立细胞系。使用这些细胞系，我们将在药理学上阻断逆转录病毒基因转移激活的单个信号通路，以评估增殖、分化和凋亡对肿瘤生长的相对贡献。这项研究的结果将为促进 MB 生长的信号转导途径提供见解。该信息可用于识别分子治疗靶点并促进化疗药物临床前测试模型的开发。