Single Cell Analyses of Neuroimmune Dysfunctions in the Thalamocortical Circuit in FTLD

FTLD 丘脑皮质回路神经免疫功能障碍的单细胞分析

基本信息

批准号：
10207374
负责人：
Eric J Huang
金额：
$ 64.6万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-15 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10207374
关键词：
Age Aging Alternative Complement Pathway Astrocytes Cell Cycle Progression Cell Death Cell Nucleus Cells Complement Complement 4b Dementia Disease Disease model Epigenetic Process Exhibits Fluorescent in Situ Hybridization Frontotemporal Lobar Degenerations Functional disorder GRN gene Gene Expression Genes Gliosis Human Image Image Analysis Knock-out Lead Link Mediating Microglia Microscopy Modeling Molecular Molecular Profiling Mus Mutation Natural Immunity Nerve Degeneration Neurodegenerative Disorders Neuroglia Neuroimmune Neurons PGRN gene Pathology Pathway Analysis Patients Phenotype Physiological Process Property Protein Deficiency Proteins RNA-Binding Proteins Reporter Resolution Role Sampling Small Nuclear RNA Surveys Synapses System Technology Testing Thalamic structure Up-Regulation Work age related aging brain base behavioral phenotyping brain tissue cell type cellular imaging cohort excitatory neuron frontal lobe glial activation inhibitory neuron innovation insight neural circuit neuron loss progenitor protein TDP-43 reconstruction response single cell analysis single molecule spatiotemporal synaptic pruning transcriptome transcriptome sequencing transcriptomics

项目摘要

PROJECT SUMMARY Aberrant glial activation is a prominent feature in neurodegenerative diseases. But, what triggers glial activation in the aging brain and how it contributes to neuronal degeneration remains unclear. The scientific premise of this proposal is based on previous studies that dominant mutations in human Progranulin gene (GRN [gene], PGRN [protein]) cause a drastic reduction in PGRN levels in CSF and brain tissues in patients with frontotemporal lobar degeneration (FTLD), leading to profound gliosis, aggregation of RNA binding protein TDP-43, and neurodegeneration. In support of this idea, our recent studies show that Grn knockout (Grn-/-) mice is a valid model that captures several key disease features in FTLD caused by GRN mutations (FTLD- GRN), including microglial activation, microglia-mediated synaptic pruning and dysfunction in the thalamocortical circuit. Our ongoing work further revealed that Grn-/- mice and FTLD-GRN patients also shows a robust astroglial activation that positively correlates with microglial activation. Similar to Grn-/- microglia, Grn-/- astrocytes exhibit an age-dependent up-regulation of innate immunity genes, including complements C3 and C4b, which together with C1qa from Grn-/- microglia, activate both classical and alternative complement pathways to promote neurodegeneration. Taken together, these results support the hypothesis that PGRN deficiency is a feasible disease model to uncover the intricate neuroimmune interactions and how perturbation to these interactions leads to neuronal degeneration. To test this hypothesis, we propose a comprehensive single cell transcriptomic approach to survey the dynamic changes of glial and neuronal cell types in the thalamocortical circuit that is most severely impacted by PGRN deficiency. This approach will provide critical insights into the intrinsic mechanism of glial activation, neuronal degeneration and neural circuit dysfunction in Grn-/- mice and in FTLD-GRN patients. This innovative strategy involves high throughput profiling of transcriptomic and physiological properties of glia and neurons using droplet-based capture technology, microscopy and dynamic imaging of cell intrinsic physiological responses. These results will provide an unprecedented resolution to directly test the hypothesis that disruptions to the dynamic neuroimmune interactions between microglia, astrocytes and neurons in the thalamocortical circuit lead to neurodegeneration in FTLD caused by PGRN deficiency.

项目摘要异常的神经胶质激活是神经退行性疾病的重要特征。但是，触发神经胶质的原因衰老大脑的激活及其如何对神经元变性有效。科学该提案的前提是基于以前的研究，即人类尿素基因中的显性突变（GRN [GENE]，PGRN [蛋白质]）患者CSF和脑组织的PGRN水平急剧降低与额颞叶变性（FTLD），导致深度神经胶质变性，RNA结合蛋白的聚集 TDP-43和神经变性。为了支持这一想法，我们最近的研究表明GRN敲除（GRN - / - ）小鼠是一个有效的模型，它捕获了由GRN突变引起的FTLD中的几个关键疾病特征（FTLD- GRN），包括小胶质细胞激活，小胶质细胞介导的突触修剪和功能障碍丘脑皮层电路。我们正在进行的工作进一步表明，GRN - / - 小鼠和FTLD-GRN患者也表明与小胶质激活正相关的强大星形胶质细胞激活。类似于GRN - / - 小胶质细胞，GRN - / - 星形胶质细胞表现出年龄依赖于先天免疫基因的上调，包括补充C3和 C4B与GRN - / - 小胶质细胞的C1QA一起激活经典和替代补体促进神经变性的途径。综上所述，这些结果支持了PGRN的假设缺乏症是一种可行的疾病模型，可揭示复杂的神经免疫相互作用以及如何扰动这些相互作用导致神经元变性。为了检验这一假设，我们提出了一个全面的单细胞转录组方法来调查神经胶质和神经元细胞类型的动态变化丘脑皮层电路受PGRN缺乏症的影响最严重。这种方法将提供关键洞悉神经胶质激活，神经元变性和神经回路功能障碍的内在机制 GRN - / - 小鼠和FTLD-GRN患者。这种创新策略涉及高吞吐量分析使用基于液滴的捕获技术的神经胶质和神经元的转录组和生理特性，细胞内在生理反应的显微镜和动态成像。这些结果将提供前所未有的决议，直接检验了破坏动态神经免疫的假设丘脑皮质回路中的小胶质细胞，星形胶质细胞和神经元之间的相互作用导致神经变性由PGRN缺乏引起的FTLD。