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: 8583509
• 负责人: XANDRA OWENS BREAKEFIELD
• 金额: $ 166.21万
• 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/8583509
• 关键词: Biogenesis; Brain; Cells; Communications Media; DNA Transposable Elements; Dependence; Development; Emerging Technologies; Environment; Epidermal Growth Factor Receptor; Evaluation; Event; Functional RNA; Genetic; Genetic Translation; Genome; Genomics; Genotype; Glioblastoma; Human; Image; Label; Malignant Neoplasms; Mediating; Methylation; MicroRNAs; Modality; Modeling; Molecular; Monitor; Mus; Normal Cell; Oncogenes; Pharmaceutical Preparations; Phenotype; Process; RNA; Radiation; Reagent; Reporter; Signal Pathway; Transfer RNA; Vesicle; Visual; brain cell; extracellular; in vivo; intravital imaging; mouse model; neoplastic cell; novel therapeutic intervention; public health relevance; 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）：我们的广义术语目标是两个方面：了解细胞外RNA（EXRNA）如何调节肿瘤环境中正常细胞的表型，并产生新的实验方式，以阐明细胞之间这种通信形式的机制。具体而言，我们将专注于了解从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。我们的总体具体目的将包括：目标1-使用现有和新兴技术来操纵这些过程。 AIM 2-人类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含量的全面表征，以及在GBM小鼠脑模型中评估EXRNA从GBM细胞到培养物中脑细胞的功能转移。 AIM 3-评估脑细胞中EXRNA的转移和命运，包括可视化电动汽车中的RNA转移，监测mRNA翻译和miRNA功能，确定可转移元素/癌基因的基因组整合，以及评估非编码EXRNAS对基因组甲基化状态的影响。 AIM 4-描述EXRNA货物组成，形成和释放动力学的依赖性是GBM基因型的函数，包括激活EGFR和PDGFRA信号通路，这是人类GBM肿瘤中最常见的两个遗传事件，以及GBM ExRNA在响应辐射和药物治疗中的GBM ExRNA的变化。 AIM 5-通过调整荧光和其他视觉标签，矢量，小鼠模型和试剂，用于监测EXRNA释放，培养和体内培养中的功能，开发ExRNA释放和吸收的调节器和记者。这些目标将由共享成像核心支持，该成像核心带有我们对细胞外囊泡的内部成像及其与大脑内源细胞的相互作用。