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.

项目概要/摘要 低级别胶质瘤 (LGG) 是一种疾病，尽管有现代护理标准，但仍经常复发或进展为高级别胶质瘤。 级神经胶质瘤，例如胶质母细胞瘤(GBM)。大多数 LGG 患者在就诊时都会出现癫痫发作，并且会复发。 越来越多的研究癫痫样活动如何影响神经胶质瘤微环境的研究表明， 这种神经胶质瘤相关癫痫（GAE）通过直接神经元-神经胶质瘤的方式促进神经胶质瘤增殖和侵袭 发信号。然而，GAE 驱动神经胶质瘤的其他机制仍不清楚。与之前的工作相比 领域，本提案重点研究 GAE 与转录组和功能变化之间的相关性 小胶质细胞是一种已知对驱动 GBM 进展至关重要的细胞类型。众所周知，小胶质细胞表现出转录组学特征 颞叶癫痫（TLE）中针对突触的变化和吞噬细胞过度活动。然而，尽管情况良好 虽然这种细胞类型在 GBM 和 TLE 中发挥的作用得到了验证，但它们在 LGG 和 TLE 中的探索仍未完全完成。 盖伊。有趣的是，研究表明人类癫痫瘤周皮层存在树突缺失，并且树突缺失 已被证明在包括癫痫在内的许多疾病中是由小胶质细胞介导的。反过来，也表明 神经元元件的吞噬作用驱动抗炎信号传导，诱导一组与小胶质细胞平行的细胞因子 显示预测 GBM 预后较差的特征。该提案将检验 GAE 相关的假设 表现出过度活跃的突触吞噬作用的小胶质细胞群的富集。此外，它将决定 突触的吞噬作用是否与提示促肿瘤发生的小胶质细胞特征的上调相关 功能。在目标 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 中差异丰富，并现场验证了我的发现。然后我将检查 GAE 的相关性 TCGA 数据集中 LGG 相关的小胶质细胞特征与临床结果。在目标 2 中，我将使用 Single Cell 光学表型和表达测序 (SCOPE-seq) 对单个小胶质细胞进行基于成像的测量 利用来自同一细胞的单细胞 RNA 测序数据来评估吞噬能力。我会检查GAE是否相关 随着小胶质细胞突触吞噬作用异常升高，将识别出表现出更大突触的小胶质细胞亚型 吞噬能力并研究突触的吞噬作用是否会驱动小胶质细胞特征的上调 表明促肿瘤功能或与预测 GBM 较差临床结果的相似功能。该项目将 阐明小胶质细胞吞噬作用和小胶质细胞转录组在 LGG 和 GAE 中如何受到不同程度的干扰， 可能揭示新的小胶质细胞治疗靶点，这些靶点可能被调节以减缓 LGG 进展。