Elucidating transcriptomic and functional heterogeneity of microglia in low-grade glioma and glioma-associated epilepsy.

阐明低级别神经胶质瘤和神经胶质瘤相关癫痫中小胶质细胞的转录组和功能异质性。

• 批准号: 10394135
• 负责人: John Tuddenham
• 金额: $ 5.18万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10394135
• 关键词: Affect; Animal Model; Anti-Inflammatory Agents; Antigen Presentation; Automobile Driving; Autopsy; Avidity; Biological Assay; Brain Neoplasms; Cell Nucleus; Cells; Characteristics; Clinical; Data; Data Set; Dendrites; Dendritic Spines; Diffuse; Disease; Elements; Epilepsy; Excision; Exhibits; Gene Expression Profile; Genes; Glioblastoma; Glioma; Goals; Growth; Heterogeneity; Homeostasis; Human; Image; Immunohistochemistry; In Situ; Incidence; Link; Macrophage Colony-Stimulating Factor Receptor; Maps; Measurement; Mediating; Microglia; Minocycline; Modeling; Modernization; Myelogenous; Neurons; Optics; Outcome; Pathologic; Patients; Phagocytes; Phagocytosis; Phenotype; Play; Population; Prevalence; Primary Brain Neoplasms; Prognosis; Recording of previous events; Recurrence; Resources; Role; Sampling; Seizures; Signal Transduction; Slice; Spatial Distribution; Structure; Suggestion; Synapses; Synaptosomes; Syndrome; Temporal Lobe Epilepsy; Testing; The Cancer Genome Atlas; Tissues; Tumor Bank; Up-Regulation; Work; base; brain parenchyma; cell type; clinically relevant; cohort; curative treatments; cytokine; gene induction; genetic signature; immune activation; immunoregulation; inhibitor; macrophage; nervous system disorder; new technology; new therapeutic target; novel; response; single-cell RNA sequencing; standard of care; therapeutic target; transcriptome; transcriptome sequencing; transcriptomics; trend; tumor; tumor growth; tumorigenic

PROJECT SUMMARY/ABSTRACT Low-grade glioma (LGG) is a disease that, despite the modern standard of care, frequently recurs or progresses to high- grade glioma, such as glioblastoma (GBM). Most LGG patients suffer from seizures on presentation and with recurrence. A growing body of work investigating how epileptiform activity affects the glioma microenvironment has suggested that this glioma-associated epilepsy (GAE) drives increased glioma proliferation and invasion by way of direct neuron-glioma signaling. However, other mechanisms by which GAE may drive glioma remain unclear. In contrast to prior work in the field, this proposal focuses on investigating the correlation between GAE and transcriptomic and functional changes in microglia, a cell type known to be critical in driving GBM progression. Microglia are well known to exhibit transcriptomic changes and phagocytic overactivity targeting synapses in temporal lobe epilepsy (TLE). However, despite the well- validated roles this cell type plays in both GBM and TLE, they have remained incompletely explored in both LGG and GAE. Intriguingly, it has been shown that there is dendritic loss in human epileptic peritumoral cortex, and dendritic loss has been shown to be microglia-mediated in many diseases, including epilepsy. In turn, it has also been shown that phagocytosis of neuronal elements drives anti-inflammatory signaling, inducing a set of cytokines that parallel microglial signatures shown to predict poorer prognosis in GBM. This proposal will test the hypothesis that GAE is associated with enrichment of microglial populations exhibiting overactive phagocytosis of synapses. Moreover, it will determine whether phagocytosis of synapses correlates with upregulation of microglial signatures suggestive of pro-tumorigenic function. In aim 1, I will use single-nucleus sequencing and immunohistochemistry or RNAscope to identify microglial subtypes differentially enriched in GAE and LGG and validate my findings in situ. I will then examine correlation of GAE and LGG-associated microglial signatures with clinical outcomes in the TCGA dataset. In aim 2, I will use Single Cell Optical Phenotyping and Expression sequencing (SCOPE-seq) to pair imaging-based measurements of single microglial phagocytic capacity with single-cell RNA-sequencing data from the same cell. I will examine whether GAE is associated with aberrantly elevated microglial synaptic phagocytosis, will identify microglial subtypes that exhibit greater synaptic phagocytic capacity and study whether phagocytosis of synapses drives upregulation of microglial signatures that suggest pro-tumorigenic function or parallel those found to predict worse clinical outcomes in GBM. This project will elucidate how microglial phagocytosis and the microglial transcriptome are differentially perturbed in LGG and GAE and may reveal novel microglial therapeutic targets that may be modulated to slow LGG progression.

项目摘要/摘要 低级神经胶质瘤（LGG）是一种疾病，尽管现代护理标准，但经常出现或发展为高 等级神经胶质瘤，例如胶质母细胞瘤（GBM）。大多数LGG患者出现癫痫发作和复发。 越来越多的工作调查了癫痫样活动如何影响神经胶质瘤微环境的工作表明， 这种与神经胶质瘤相关的癫痫（GAE）通过直接神经元神经胶质瘤驱动神经胶质瘤增殖和侵袭 信号。但是，GAE可能驱动神经胶质瘤的其他机制尚不清楚。与先前的工作相反 该领域，该提案的重点是研究GAE与转录组和功能变化之间的相关性 小胶质细胞，一种已知在驱动GBM进展至关重要的细胞类型。小胶质细胞众所周知，可以表现出转录组 颞叶癫痫（TLE）中靶向突触的变化和吞噬细胞过度活动。但是，尽管有很好的 经过验证的角色，这种细胞类型在GBM和TLE中都扮演的角色，在LGG和 GAE。有趣的是，已经表明，人癫痫周围皮层和树突状损失有树突状损失 已显示在包括癫痫在内的许多疾病中是小胶质细胞介导的。反过来，也已经表明 神经元元素的吞噬作用驱动抗炎信号传导，诱导一组平行小胶质细胞因子 签名预测GBM的预后较差。该提案将检验GAE与 富集着表现出突触过度活跃吞噬作用的小胶质细胞。而且，它将确定 突触的吞噬作用是否与小胶质细胞的上调有关 功能。在AIM 1中，我将使用单核测序和免疫组织化学或RNASCOPE来识别小胶质细胞 亚型在GAE和LGG中差异化，并在原位验证我的发现。然后，我将检查GAE的相关性 与LGG相关的小胶质细胞特征在TCGA数据集中具有临床结果。在AIM 2中，我将使用单个单元格 光学表型和表达测序（范围seq），以对单个小胶质细胞的基于成像的测量 带有来自同一细胞的单细胞RNA测序数据的吞噬能力。我将检查GAE是否相关 小胶质细胞突触吞噬作用异常升高，将鉴定出表现出更大突触的小胶质细胞类型 吞噬能力并研究突触的吞噬作用是否会驱动小oglial特征的上调 提示促肿瘤功能或与发现可以预测GBM中临床结果较差的相关功能。这个项目将 阐明如何在LGG和GAE中差异化小胶质细胞增多症和小胶质细胞转录组。 可能会揭示新型的小胶质细胞治疗靶标，这些靶标可能会调节以减慢LGG进展。