Project 1 Extracellular Vesicles (EVs) Role in GBM Pathobiology

项目 1 细胞外囊泡 (EV) 在 GBM 病理学中的作用

• 批准号: 10213217
• 负责人: XANDRA OWENS BREAKEFIELD
• 金额: $ 28.44万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10213217
• 关键词: Address; Biological Markers; Biology; Blood specimen; Brain; Brain Glioblastoma; Brain Neoplasms; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell membrane; Cells; Cerebrospinal Fluid; Clinical Trials; Collaborations; Communication; Cytotoxic T-Lymphocytes; Development; Drug resistance; Environment; Epidermal Growth Factor Receptor; Epithelial; Equation; Gene Expression Profile; Genes; Genetic; Genotype; Glioblastoma; Glioma; Goals; Heterogeneity; Human; Immune; Immune response; Immunosuppression; Immunotherapy; Infusion procedures; Label; Language; Letters; Ligands; Luciferases; Malignant - descriptor; Malignant Neoplasms; Mediating; Mesenchymal; Messenger RNA; MicroRNAs; Microglia; Modeling; Modification; Monitor; Mus; Normal Cell; Nucleic Acids; Patients; Pharmacotherapy; Phenotype; Plasma; Population; Process; Property; Protein Analysis; Proteins; RNA; Regulatory T-Lymphocyte; Reporting; Reproducibility; Resistance; Role; Solid Neoplasm; T-Lymphocyte; Therapeutic; Transfer RNA; Tumor Tissue; Tumor-associated macrophages; United States National Institutes of Health; biomaterial compatibility; capsule; cell type; checkpoint inhibition; cytotoxic; driving force; epithelial to mesenchymal transition; extracellular vesicles; immunological status; implantation; improved; macrophage; mouse model; neoplastic cell; patient derived xenograft model; preclinical evaluation; programmed cell death ligand 1; response; small molecule; stem; stem-like cell; success; therapy resistant; transcription factor; transcriptome; transcriptome sequencing; tumor; tumor growth; tumor microenvironment; tumor progression; uptake; vector; vesicular release

Glioblastomas (GBMs) are one of the most difficult cancers to treat. Their success in defying therapy relies in large part on their dynamic cellular heterogeneity and on their ability to subvert their environment, in particular to create an immune suppressive milieu. This project will address the role of the newly discovered communication vehicles in cancer, extracellular vesicles (EVs) which can carry RNA and protein as informational molecules. Our goal is to evaluate to what extent EVs produced by these tumors contribute to an immune suppressive environment through their interaction with tumor-associated macrophages and microglia (TAMs), and to what extent EVs produced by different genetic subtypes of gliomas, and the stem-like cells within these tumors, underlie to their ability to re-define themselves and escape therapy. In Aim 1 we will assess EV-mediated information transfer between glioma tumors and microglia/macrophages in the tumor microenvironment and its effect on immune suppression. In Aim 2 we will evaluate the role of EVs in modulating GBM stem-like cell (GSC) phenotypes and therapeutic resistance. Studies will use syngeneic mouse glioma models as well as human GBM stem-like neurosphere cultures and patient-derived xenograft in mouse models developed with Core C. We will generate an array of vectors to label cells with multiple luciferases and fluorescent proteins, as well as create capsules to contain tumor cells while allowing release of EVs and/or small molecules within the brain. FACS analysis and RNAseq will be used to define the transcriptome of different cell populations in the brain. Biofluids from mice, as well as from GBM patients (Core B), will be sent to Project 2 for protein analysis. Correlative analysis of mRNA/ protein contents and GBM genotypes will be assessed, as well as how EV contents reflect changes in GBM subtype and resistance to therapy. Our findings will be integrated with those of Project 3 in the context of preclinical evaluation of Gene- Mediated Cytotoxic Immunotherapy (GMIC) combined with immune checkpoint inhibition. Organizational oversight of rigor and reproducibility will be provided by Core A. Studies in this project will serve to inform the development of the projected clinical trial by elucidating how EVs influence and report on GBM status, and how they can be manipulated to improve this type of immunotherapy in the context of the GBM brain tumors.

胶质母细胞瘤(GBM)是最难治疗的癌症之一。他们违抗治疗的成功依赖于 很大程度上取决于它们动态的细胞异质性，特别是它们颠覆环境的能力 以创造一种免疫抑制的环境。这个项目将解决新发现的 癌症中的通讯载体，细胞外小泡(EVS)可以携带RNA和蛋白质作为 信息分子。我们的目标是评估这些肿瘤产生的EV在多大程度上有助于 与肿瘤相关巨噬细胞和小胶质细胞相互作用的免疫抑制环境 (TAMS)，以及不同遗传亚型的胶质瘤和干细胞在多大程度上产生EVS 在这些肿瘤中，基础是他们重新定义自己和逃避治疗的能力。在目标1中，我们将 评估EV介导的肿瘤与肿瘤内小胶质/巨噬细胞之间的信息传递 微环境及其对免疫抑制的影响。在目标2中，我们将评估电动汽车在 调节基底膜干细胞(GSC)表型和治疗耐药性。研究将使用同基因 小鼠胶质瘤模型以及人GBM干样神经球培养和患者来源的异种移植 用CoreC开发的小鼠模型。我们将生成一个载体数组来标记细胞 荧光素酶和荧光蛋白，以及创建胶囊以包含肿瘤细胞，同时允许释放 EVS和/或大脑中的小分子。将使用FACS分析和RNAseq来定义 大脑中不同细胞群的转录组。来自小鼠和GBM患者的生物液(Core B)，将被送到项目2进行蛋白质分析。MRNA/蛋白质含量与基底膜的相关性分析 将评估基因类型，以及EV含量如何反映GBM亚型的变化和对 心理治疗。我们的发现将与项目3的结果结合起来，在基因-临床前评估的背景下- 介导性细胞毒免疫治疗(GMIC)与免疫检查点抑制相结合。组织 严谨性和重复性的监督将由核心A提供。本项目的研究将有助于向 通过阐明EVS如何影响和报告GBM状态以及如何进行计划的临床试验的发展 在GBM脑瘤的情况下，可以操纵它们来改进这种类型的免疫治疗。