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Sonic Hedghog Signaling in Cerebellum Development and Medulloblastoma

小脑发育和髓母细胞瘤中的 Sonic Hedghog 信号传导

基本信息

批准号：
7555050
负责人：
Susana Parathath
金额：
$ 5.72万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-12-24 至 2009-12-23
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7555050
关键词：
Ablation Adverse effects Behavioral Biological Models Birth Brain Breast Cancer Cell Bromodeoxyuridine Cell Cycle Progression Cells Cerebellum Childhood Brain Neoplasm Core Facility Cytoplasmic Granules Data Defect Development Equilibrium Erinaceidae Failure Fellowship Flow Cytometry Genetic Genetic Transcription Goals Human IGF1 gene Impaired cognition In Vitro Insulin-Like Growth Factor Receptor Knockout Mice Lead Life Long-Term Survivors Maintenance Mass Spectrum Analysis Mediating Memorial Sloan-Kettering Cancer Center Mentors Messenger RNA Modeling Mus Mutant Strains Mice N-myc Proto-Oncogenes Neuroblastoma Neurologic Neurons Operative Surgical Procedures Pathway interactions Patients Pattern Phase Play Polyribosomes Production Proliferating Proliferation Marker Proteins Radiation Regulation Retroviridae Role Signal Pathway Signal Transduction Somatomedins Source Structure Syndrome Techniques Testing Therapeutic Transgenic Mice Translations Up-Regulation Work cancer therapy cell type chemotherapy cyclin D2 in vivo inhibitor/antagonist insight insulin receptor substrate 1 protein interest medulloblastoma migration neoplastic cell novel postnatal precursor cell programs promoter protein degradation recombinase retroviral-mediated small hairpin RNA small molecule transcription factor tumor

项目摘要

DESCRIPTION (provided by applicant): The cerebellum is a brain structure responsible for integrating and coordinating neurological inputs from several sources in order to maintain equilibrium and balance. It is an interesting brain structure in that it develops after birth. One of the cell types within the cerebellum that is critical for proper postnatal development are cerebellar granule neuron precursors (CGNP). During the first two weeks of life in mice these cells undergo a rapid expansion phase. Failure of CGNPs to exit this high proliferate program has been suggested to lead to medulloblastoma formation, the most common form of pediatric brain tumor. Medulloblastoma is relatively responsive to traditional cancer treatments, including surgery, chemotherapy and radiation. However long-term survivors often suffer from life-long developmental, behavioral, and cognitive disturbances. These devastating side effects underscore the need to understand the basic mechanisms underlying medulloblastoma, so novel treatments can be developed to specifically target tumor cells without damaging the developing brain. The long-term objectives of this project is to identify nodes of cross-talk between Sonic hedgehog and IGF signaling pathways both of which have been implicated in medulloblastoma. Greater understanding of these pathways may lead to the development of small molecule treatments reducing the need for radiation and chemotherapy in these young patients. In order to accomplish this goal we have developed an interdisciplinary proposal incorporating various techniques and models. Our preliminary data indicate that Shh signaling increases IRS1 protein levels, a scafold directly downstream of the IGF receptor, but not IRS1 mRNA in cultured CGNPs. In aim 1 we will test the hypothesis that Shh proliferative signaling in CGNPs positively regulates IGF pathway activity through up-regulation of IRS1 protein. We will look at IRS1 protein turnover as well as polysome analysis including microarray and mass spectrometry analysis. In aim 2 we will use retroviral mediated over-expression of IRS1 and lentivirally-delivered shRNAto determine if IRS1 is necessary/sufficient for Shh mediated CGNP proliferation in culture. In aim 3 we will determine the role of IRS1 in vivo by analyzing CGNP proliferation and cerebella development in mice lacking IRS1 either constitutively or conditionally in the cerebellum. In conjunction with MSKCC Mouse Genetics core facility we will develop mice conditionally lacking IRS1 in the cerebellum. We will analyze the role of CGNP proliferation using in vivo as well as in vitro by culturing CGNPs and treating with Cre to remove IRS1.

描述（由申请人提供）：小脑是一种大脑结构，负责整合和协调来自多个来源的神经输入，以维持平衡和平衡。这是一个有趣的大脑结构，因为它是在出生后发育的。小脑内对出生后正常发育至关重要的细胞类型之一是小脑颗粒神经元前体 (CGNP)。在小鼠生命的前两周，这些细胞经历快速扩增阶段。有人认为，CGNP 未能退出这种高增殖程序会导致髓母细胞瘤形成，这是最常见的儿科脑肿瘤形式。髓母细胞瘤对传统癌症治疗（包括手术、化疗和放疗）相对敏感。然而，长期幸存者往往会遭受终生的发育、行为和认知障碍。这些毁灭性的副作用强调需要了解髓母细胞瘤的基本机制，因此可以开发新的治疗方法来专门针对肿瘤细胞而不损害发育中的大脑。该项目的长期目标是确定 Sonic Hedgehog 和 IGF 信号通路之间的串扰节点，这两种信号通路均与髓母细胞瘤有关。对这些途径的更多了解可能会导致小分子治疗的发展，从而减少这些年轻患者对放射和化疗的需求。为了实现这一目标，我们制定了一项包含各种技术和模型的跨学科提案。我们的初步数据表明，Shh 信号传导会增加 IRS1 蛋白水平（IGF 受体的直接下游支架），但不会增加培养的 CGNP 中的 IRS1 mRNA。在目标 1 中，我们将检验以下假设：CGNP 中的 Shh 增殖信号通过上调 IRS1 蛋白来正向调节 IGF 通路活性。我们将研究 IRS1 蛋白质周转以及多核糖体分析，包括微阵列和质谱分析。在目标 2 中，我们将使用逆转录病毒介导的 IRS1 过表达和慢病毒递送的 shRNA 来确定 IRS1 对于培养中 Shh 介导的 CGNP 增殖是否是必要/充分的。在目标 3 中，我们将通过分析小脑中组成性或条件性缺乏 IRS1 的小鼠的 CGNP 增殖和小脑发育来确定 IRS1 在体内的作用。与 MSKCC 小鼠遗传学核心设施相结合，我们将开发小脑有条件地缺乏 IRS1 的小鼠。我们将通过培养 CGNP 并用 Cre 处理去除 IRS1，在体内和体外分析 CGNP 增殖的作用。