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: 8464013
• 负责人: DANIEL WEBSTER FULTS
• 金额: $ 23.05万
• 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 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8464013
• 关键词: 1-Phosphatidylinositol 3-Kinase; Abbreviations; Achievement; Apoptosis; Automobile Driving; Avian Leukosis Virus; BCL2 gene; Biological Models; Brain Injuries; Cell Proliferation; Cell Surface Receptors; Cell Survival; Cells; Cerebellum; Cerebrospinal Fluid; Child; Cytoplasmic Granules; Development; Ectopic Expression; Erinaceidae; Gene Transfer; Genes; Genetically Engineered Mouse; Growth; Growth Factor; Health; Hematoxylin and Eosin Staining Method; Hepatocyte Growth Factor; Human; Insulin-Like Growth Factor II; Long Terminal Repeats; Malignant neoplasm of brain; Modeling; Molecular Target; Monoclonal Antibodies; Monoclonal Antibody Therapy; Mus; Neoplasm Metastasis; Neurofilament Proteins; Neurological outcome; Neurons; Oncogene Proteins; Operative Surgical Procedures; Pathogenesis; Pathway interactions; Patients; Phenotype; Play; Preclinical Testing; Prognostic Factor; Protein Tyrosine Kinase; Proteins; Proto-Oncogene Protein c-met; RNA Splicing; Radiation; Research; Retroviral Vector; Retroviridae; Ribosomal Protein S6; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Site; Sleeping Beauty; Somatic Cell; Spinal; Stimulus; Subgroup; Survival Rate; System; Transgenes; Transgenic Mice; Translating; Treatment Protocols; Vertebral column; chemotherapy; density; enhancing factor; human disease; improved; intraperitoneal; medulloblastoma; meetings; mouse model; neoplastic cell; nerve stem cell; nestin protein; neurotoxicity; partial response; postnatal; programs; promoter; receptor; smoothened signaling pathway; subcutaneous; therapeutic target; treatment response; tumor; tumor growth; vector

DESCRIPTION (provided by applicant): Medulloblastomas (MBs) are malignant brain tumors that arise by transformation of neural progenitor cells in the cerebellum in children. Aggressive treatment approaches combining surgery, craniospinal radiation, and chemotherapy result in 5-year survival rates exceeding 70%. Treatment- related neurotoxicity has created a critical need to identify signaling molecules that can be targeted therapeutically to maximize tumor growth suppression and minimize collateral brain damage. The overall objective of this research is to use a mouse model of MB, which we developed using the RCAS/tv-a gene transfer system, as a preclinical testing platform for molecular targeted MB therapy. This experimental system uses a retroviral vector (RCAS) derived from avian leukosis virus and a transgenic mouse line that expresses the retrovirus receptor under control of the Nestin gene promoter, which is active in neural progenitor cells during normal cerebellar development. Using this system, we showed that ectopic expression of Sonic Hedgehog (Shh) in the postnatal cerebellum induces MBs in mice. Furthermore, we identified proteins belonging to different functional classes that cooperate with Shh to enhance MB formation. These enhancing factors are (a) Myc oncoproteins, which stimulate proliferation of neural progenitors during normal development, (b) Bcl-2, which potently inhibits apoptosis, (c) insulin-like growth factor-II, which concomitantly stimulates proliferation and blocks apoptosis by activating the phosphatidylinositol 3-kinase (PI3K) signal transduction pathway, and (d) hepatocyte growth factor (HGF), a growth factor with pleiotropic effects on tumor growth. The fact that all of these proteins are highly expressed in human MBs indicates that their tumor-promoting activity in mice accurately reflects the pathogenesis of the human disease. Moreover, these proteins and their downstream signaling molecules can be considered therapeutic targets. Specific aim 1 is to determine whether inhibiting Shh signaling cooperates with blockade of HGF signaling to enhance treatment response in mice bearing Shh?induced MBs. Specific aim 2 is to determine whether MB growth suppression by HGF monoclonal antibody therapy can be enhanced by pharmacologic inhibition of the HGF receptor c- Met or downstream PI3K signaling. Specific aim 3 is to use the RCAS/tv-a system to identify genes that cause Shh-induced MBs to metastasize to the spine. Spinal metastasis is a highly unfavorable prognostic factor for patients with MBs. Achievement of these aims will make it possible to translate the mechanistic discoveries to molecular targeted therapies for children with MB.

描述（由申请人提供）：髓母细胞瘤（MB）是由儿童小脑神经祖细胞转化引起的恶性脑肿瘤。结合手术、颅脊髓放疗和化疗的积极治疗方法可使 5 年生存率超过 70%。与治疗相关的神经毒性迫切需要识别可靶向治疗的信号分子，以最大限度地抑制肿瘤生长并最大限度地减少附带的脑损伤。本研究的总体目标是利用我们利用 RCAS/tv-a 基因转移系统开发的 MB 小鼠模型作为分子靶向 MB 治疗的临床前测试平台。该实验系统使用源自禽白血病病毒的逆转录病毒载体（RCAS）和转基因小鼠系，该转基因小鼠系在巢蛋白基因启动子的控制下表达逆转录病毒受体，该受体在正常小脑发育过程中的神经祖细胞中活跃。使用该系统，我们发现 Sonic Hedgehog (Shh) 在出生后小脑中的异位表达可诱导小鼠产生 MB。此外，我们还鉴定了属于不同功能类别的蛋白质，它们与 Shh 合作增强 MB 形成。这些增强因子是 (a) Myc 癌蛋白，在正常发育过程中刺激神经祖细胞增殖；(b) Bcl-2，有效抑制细胞凋亡；(c) 胰岛素样生长因子-II，通过激活磷脂酰肌醇 3 激酶 (PI3K) 信号转导途径，同时刺激增殖并阻止细胞凋亡；(d) 肝细胞生长因子（HGF），一种对肿瘤生长具有多效作用的生长因子。所有这些蛋白质在人类 MB 中高表达的事实表明它们在小鼠中的促肿瘤活性准确地反映了人类疾病的发病机制。此外，这些蛋白质及其下游信号分子可以被视为治疗靶点。具体目标1是确定抑制Shh信号传导是否与阻断HGF信号传导协同作用，以增强携带Shh？诱导的MB的小鼠的治疗反应。具体目标 2 是确定 HGF 单克隆抗体治疗对 MB 生长的抑制是否可以通过 HGF 受体 c-Met 或下游 PI3K 信号传导的药理学抑制来增强。具体目标 3 是使用 RCAS/tv-a 系统来识别导致 Shh 诱导的 MB 转移至脊柱的基因。脊柱转移对于MB患者来说是一个非常不利的预后因素。这些目标的实现将使将机制发现转化为儿童 MB 的分子靶向治疗成为可能。

项目成果

Clonal selection drives genetic divergence of metastatic medulloblastoma.

克隆选择驱动转移性髓母细胞瘤的遗传差异。

• DOI: 10.1038/nature10825
• 发表时间: 2012-02-15
• 期刊: NATURE
• 影响因子: 64.8
• 作者: Wu, Xiaochong;Northcott, Paul A.;Dubuc, Adrian;Dupuy, Adam J.;Shih, David J. H.;Witt, Hendrik;Croul, Sidney;Bouffet, Eric;Fults, Daniel W.;Eberhart, Charles G.;Garzia, Livia;Van Meter, Timothy;Zagzag, David;Jabado, Nada;Schwartzentruber, Jeremy;Majewski, Jacek;Scheetz, Todd E.;Pfister, Stefan M.;Korshunov, Andrey;Li, Xiao-Nan;Scherer, Stephen W.;Cho, Yoon-Jae;Akagi, Keiko;MacDonald, Tobey J.;Koster, Jan;McCabe, Martin G.;Sarver, Aaron L.;Collins, V. Peter;Weiss, William A.;Largaespada, David A.;Collier, Lara S.;Taylor, Michael D.
• 通讯作者: Taylor, Michael D.