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: 10645000
• 负责人: John Tuddenham
• 金额: $ 5.27万
• 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/10645000
• 关键词: Ablation; Affect; Animal Model; Anti-Inflammatory Agents; Antigen Presentation; Automobile Driving; Autopsy; Avidity; Biological Assay; Brain Neoplasms; Categories; 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; Infiltration; Invaded; Link; Macrophage; Macrophage Colony-Stimulating Factor Receptor; Maps; Measurement; Mediating; Microglia; Minocycline; Modeling; Modernization; Myelogenous; Neuroglia; Neurons; Optics; Outcome; Pathologic; Patients; Phagocytes; Phagocytosis; Phenotype; Play; Population; Prevalence; Primary Brain Neoplasms; Prognosis; Proliferating; Recording of previous events; Recurrence; Resources; Role; Sampling; Seizures; Signal Transduction; Slice; Spatial Distribution; Structure; Synapses; Synaptosomes; Syndrome; Temporal Lobe Epilepsy; Testing; The Cancer Genome Atlas; Tissues; Tumor Bank; Up-Regulation; Work; brain parenchyma; cell type; clinically relevant; cohort; curative treatments; cytokine; epileptiform; gene induction; genetic signature; immune activation; immunoregulation; inhibitor; nervous system disorder; new technology; new therapeutic target; novel; response; single nucleus RNA-sequencing; single-cell RNA sequencing; standard of care; therapeutic target; transcriptome; 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.

项目总结/摘要 低级别胶质瘤（LGG）是一种疾病，尽管现代标准的护理，经常复发或进展到高级别， 级别胶质瘤，如胶质母细胞瘤（GBM）。大多数LGG患者在就诊时癫痫发作并复发。 越来越多的研究癫痫样活动如何影响胶质瘤微环境的工作表明， 这种胶质瘤相关性癫痫（GAE）通过直接神经元胶质瘤 发信号。然而，GAE可能驱动胶质瘤的其他机制仍不清楚。与先前的工作相比， 该建议的重点是研究GAE与转录组学和功能变化之间的相关性， 小胶质细胞，一种已知在驱动GBM进展中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）中靶向突触的变化和吞噬细胞过度活跃。然而，尽管好- 虽然这种细胞类型在GBM和TLE中发挥的作用已得到证实，但它们在LGG和TLE中的研究仍不完全。 GAE有趣的是，已经表明，在人类癫痫瘤周皮层中存在树突缺失， 在包括癫痫在内的许多疾病中，都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，诱导一系列与小胶质细胞平行的细胞因子 显示的特征预测GBM的预后较差。该提案将检验GAE与以下假设有关： 小胶质细胞群的富集表现出突触的过度活跃的吞噬作用。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信号的上调相关 功能目的一：利用单核细胞测序和免疫组化或RNA显微镜技术鉴定小胶质细胞 在GAE和LGG中差异富集的亚型，并验证了我的原位研究结果。然后，我将检查GAE的相关性 和LGG相关的小胶质细胞特征与TCGA数据集中的临床结果。在目标2中，我将使用单细胞 光学表型和表达测序（SCOPE-seq），用于配对单个小胶质细胞的基于成像的测量 吞噬能力与来自相同细胞的单细胞RNA测序数据。我会检查GAE是否与 与异常升高的小胶质细胞突触吞噬作用，将确定小胶质细胞亚型，表现出更大的突触吞噬作用， 吞噬能力，并研究突触的吞噬作用是否驱动小胶质细胞信号的上调， 提示促肿瘤发生功能或发现预测GBM中更差临床结果的平行功能。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中的差异干扰， 可能揭示了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。