exRNA released by glioblastoma alters brain microenvironment

胶质母细胞瘤释放的 exRNA 改变大脑微环境

• 批准号: 9252083
• 负责人: XANDRA OWENS BREAKEFIELD
• 金额: $ 33.8万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9252083
• 关键词: Biogenesis; Brain; Cells; Communications Media; DNA Transposable Elements; Dependence; Development; Emerging Technologies; Environment; Epidermal Growth Factor Receptor; Evaluation; Event; Genetic; Genetic Translation; Genome; Genomics; Genotype; Glioblastoma; Health; Human; Image; Label; Malignant Neoplasms; Mediating; Methylation; MicroRNAs; Modality; Modeling; Molecular; Monitor; Mus; Normal Cell; Oncogenes; Pharmacotherapy; Phenotype; Process; RNA; Radiation therapy; Reagent; Reporter; Signal Pathway; Transfer RNA; Untranslated RNA; Visual; brain cell; extracellular; extracellular vesicles; in vivo; intravital imaging; mouse model; neoplastic cell; novel therapeutic intervention; radiation response; tumor; tumor growth; uptake; vector

DESCRIPTION (provided by applicant): Our broad term objective is two-fold: to understand how extracellular RNA (exRNA) modulates the phenotype of normal cells in the tumor environment and to generate new experimental modalities that can elucidate mechanisms underlying this form of communication among cells. Specifically we will focus on understanding how exRNA released from glioblastoma cells modulates the phenotype of normal cells in the vicinity of the tumor. Our overall specific aims will encompass: Aim 1 - Elucidation of basic molecular and cellular mechanisms of exRNA biogenesis by GBM cells, and uptake and function in normal brain cells, using existing and emerging technologies to manipulate these processes. Aim 2 - Full characterization of the exRNA content and intracellular RNA content of human GBM cells, as well as the intracellular RNA content of normal brain cells, and evaluation of functional transfer of exRNAs from GBM cells to brain cells in culture and in GBM mouse brain models. Aim 3 - Evaluation of transfer and fate of exRNA in brain cells, including visualizing RNA transfer in EVs, monitoring mRNA translation and miRNA functions, determining possible genomic integration of transposable elements/oncogenes, and evaluation of effects of non-coding exRNAs on status of genome methylation. Aim 4 - Description of the dependence of exRNA cargo composition, formation and release dynamics as a function of GBM genotype, including activation of EGFR and PDGFRa signaling pathways, the two most common genetic events in human GBM tumors, as well as changes in GBM exRNA in response to radiation and drug treatment. Aim 5 - Development of regulators and reporters of exRNA release and uptake by tailoring fluorescent and other visual labels, vectors, mouse models and reagents for broad applications in monitoring exRNA release, uptake and function in culture and in vivo. These aims will be supported by a shared imaging core carrying our intravital imaging of extracellular vesicles and their interaction with endogenous cells in the brain.

描述（由申请人提供）：我们的广义目标有两个：了解细胞外 RNA (exRNA) 如何调节肿瘤环境中正常细胞的表型，并产生新的实验模式，以阐明这种形式的通信背后的机制。细胞。具体来说，我们将重点了解胶质母细胞瘤细胞释放的 exRNA 如何调节肿瘤附近正常细胞的表型。我们的总体具体目标将包括： 目标 1 - 阐明 GBM 细胞 exRNA 生物发生的基本分子和细胞机制，以及正常脑细胞的摄取和功能，使用现有和新兴技术来操纵这些过程。目标 2 - 全面表征人 GBM 细胞的 exRNA 含量和细胞内 RNA 含量，以及正常脑细胞的细胞内 RNA 含量，并评估 exRNA 从 GBM 细胞到培养物和 GBM 小鼠脑中脑细胞的功能转移模型。目标 3 - 评估脑细胞中 exRNA 的转移和命运，包括可视化 EV 中的 RNA 转移、监测 mRNA 翻译和 miRNA 功能、确定转座元件/癌基因可能的基因组整合，以及评估非编码 exRNA 对脑细胞状态的影响基因组甲基化。目标 4 - 描述 exRNA 货物组成、形成和释放动力学与 GBM 基因型的关系，包括 EGFR 和 PDGFRa 信号通路的激活（人类 GBM 肿瘤中两个最常见的遗传事件）以及 GBM exRNA 的变化对放射和药物治疗的反应。目标 5 - 通过定制荧光和其他视觉标记、载体、小鼠模型和试剂来开发 exRNA 释放和摄取的调节剂和报告基因，以广泛应用于监测培养物和体内的 exRNA 释放、摄取和功能。这些目标将得到一个共享成像核心的支持，该核心承载着细胞外囊泡的活体成像及其与大脑内源性细胞的相互作用。