Microglia contribution to disease pathogenesis in C9orf72 ALS/FTD

小胶质细胞对 C9orf72 ALS/FTD 疾病发病机制的贡献

基本信息

批准号：
10477246
负责人：
Rita Sattler
金额：
$ 83.14万
依托单位：
ST. JOSEPH'S HOSPITAL AND MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10477246
关键词：
ALS patients Affect Alzheimer&apos s Disease Alzheimer&apos s disease related dementia Amyotrophic Lateral Sclerosis Area Astrocytes Autopsy Brain Brain region C9ORF72 Candidate Disease Gene Cell Differentiation process Cell Line Cells Clinical Coculture Techniques Communication Data Data Set Dementia Dementia With Amyotrophic Lateral Sclerosis Department of Defense Development Dipeptides Disease Drug Targeting Etiology Executive Dysfunction Expression Profiling Frontotemporal Dementia Functional disorder Funding Future Gene Expression Gene Expression Profile Gene Expression Profiling Gene Proteins Genes Goals Hot Spot Human Immune Immunohistochemistry Impaired cognition In Vitro Investigation Knowledge Label Laboratories Lysosomes Maintenance Measures Microglia Modeling Molecular Morphology Motor Neurons Mutation Nerve Degeneration Neurodegenerative Disorders Neurofibrillary Tangles Neuronal Dysfunction Neurons Outcome PGRN gene Pathogenesis Pathologic Pathology Patients Phagocytes Phenotype Play Prefrontal Cortex Process Property Prosencephalon Proteins RNA Regulation Reporting Role Small Nuclear RNA Specificity Symptoms Synapses System Time Tissue-Specific Gene Expression Tissues Validation Variant cell type chemokine clinical diagnosis cytokine density disease phenotype drug discovery executive function frontotemporal lobar dementia-amyotrophic lateral sclerosis genetic profiling induced pluripotent stem cell molecular phenotype neuron loss neuronal excitability neuronal survival novel novel therapeutics patient population patient subsets protein TDP-43 protein expression relating to nervous system single-cell RNA sequencing stem cell differentiation transcriptome transcriptome sequencing

项目摘要

Project Abstract The role of microglia in the C9orf72 (C9) amyotrophic lateral sclerosis (ALS)/ frontotemporal dementia (FTD) disease spectrum remains poorly understood. Early investigations found that microglia activation was significantly higher in ALS with dementia and impaired executive function, suggesting that microglia activation correlates with FTD-like symptoms in ALS. More recent neuropathologic examinations of microglia in FTLD patient autopsy brains with mutations in progranulin versus C9orf72 concluded that the observed microglia dysfunction was different between the two genetically different patient subgroups suggesting specificity of microglia dysfunction depending on the etiology of the patient population. One interesting aspect of microglia- neuron communication is the role of microglia in the maintenance and refinement of synaptic networks through the selective pruning of synapses, which occurs predominantly during development but has been shown to also be triggered in Alzheimer's disease (AD) and related dementias, including FTD. The degree of synapse loss in AD strongly correlates with cognitive decline, even more than the amount of plaque, tangles or neuronal loss, and a recent study of ALS postmortem tissue confirmed increased synapse loss in the prefrontal cortex of patients with reported cognitive impairments. Our laboratory has preliminary data supporting the hypothesis that there is an altered neural-immune interaction in the cortical forebrain regions of C9orf72 patients with confirmed FTD in which microglia and neurons modify each other's function. Using patient-derived hiPSC microglia and cortical neurons, we are able to show that C9 patient-derived hiPSC microglia mono-cultures do have intrinsic phenotypes, including altered gene profiles, phagocytic activities and lysosomal function. Most interestingly, preliminary data suggests that C9 microglia do regulate neuronal excitability and survival of C9 iPSC neurons. To further investigate the role and contribution of microglia in C9 cortical degeneration, we propose to thoroughly investigate the intrinsic properties of C9 hiPSC-microglia (from all patient subgroups: FTD, FTD/ALS, ALS; Aim1). For the first time, we will then co-culture these microglia with C9 and healthy control hiPSC cortical neurons to better understand the co-regulation between these two cell types (Aim 2). Finally, in the third aim, we will study microglia activation and pathology in C9 patient postmortem autopsy tissue. This will include cell-type specific genetic profiling from existing snRNA seq data sets, immunohistochemistry of microgliosis and multi-label immunostaining for microglial-specific candidate genes/proteins in conjunction with C9 neuronal disease pathology markers (TDP-43 and C9 DPRs) to gain novel knowledge on whether microglia are preferentially altered in close vicinity to neuronal pathologies.

项目摘要小胶质细胞在 C9orf72 (C9) 肌萎缩侧索硬化症 (ALS)/额颞叶痴呆 (FTD) 中的作用疾病谱仍然知之甚少。早期研究发现小胶质细胞的激活伴有痴呆和执行功能受损的 ALS 患者显着较高，表明小胶质细胞激活与 ALS 中的 FTD 样症状相关。 FTLD 中小胶质细胞的最新神经病理学检查对具有颗粒体蛋白前体与 C9orf72 突变的患者尸检大脑得出的结论是，观察到的小胶质细胞两个基因不同的患者亚组之间的功能障碍有所不同，这表明功能障碍的特异性小胶质细胞功能障碍取决于患者群体的病因。小胶质细胞的一个有趣的方面是—— 神经元通讯是小胶质细胞通过以下方式维持和细化突触网络的作用突触的选择性修剪，主要发生在发育过程中，但也已被证明会引发阿尔茨海默病 (AD) 和相关痴呆症，包括 FTD。突触损失程度 AD 与认知能力下降密切相关，甚至超过斑块、缠结或神经元损失的数量，最近对 ALS 死后组织的研究证实，前额叶皮层的突触损失增加报告有认知障碍的患者。我们的实验室有初步数据支持以下假设： C9orf72 患者的皮质前脑区域的神经免疫相互作用发生了改变证实了 FTD，其中小胶质细胞和神经元互相修改了彼此的功能。使用患者来源的 hiPSC 小胶质细胞和皮质神经元，我们能够证明 C9 患者来源的 hiPSC 小胶质细胞单一培养物确实具有内在的表型，包括改变的基因谱、吞噬活性和溶酶体功能。最多有趣的是，初步数据表明 C9 小胶质细胞确实调节神经元兴奋性和 C9 的存活 iPSC 神经元。为了进一步研究小胶质细胞在 C9 皮质变性中的作用和贡献，我们提议彻底研究 C9 hiPSC-小胶质细胞的内在特性（来自所有患者亚组：FTD、 FTD/ALS、ALS；目标1）。然后，我们将首次将这些小胶质细胞与 C9 和健康对照共培养 hiPSC 皮质神经元，以更好地了解这两种细胞类型之间的共同调节（目标 2）。最后，在第三个目标，我们将研究 C9 患者死后尸检组织中的小胶质细胞激活和病理学。这将包括来自现有 snRNA seq 数据集的细胞类型特异性遗传分析、免疫组织化学小胶质细胞增生和小胶质细胞特异性候选基因/蛋白质的多标记免疫染色 C9 神经元疾病病理学标记物（TDP-43 和 C9 DPR）以获得有关小胶质细胞是否存在的新知识优先在靠近神经元病理的地方改变。