A transcriptomic atlas of immune cells in a model of synucleinopathy

突触核蛋白病模型中免疫细胞的转录组图谱

• 批准号: 9808086
• 负责人: SERGE E PRZEDBORSKI
• 金额: $ 12.15万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9808086
• 关键词: Adult; Alleles; Alzheimer' s Disease; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Area; Atlases; Biological Markers; Blood; Brain; Brain region; Cell Count; Cells; Characteristics; Chronic; Computational Technique; Cost of Illness; Data; Dementia; Development; Disease; Environment; Exhibits; Genes; Goals; Heterogeneity; Immune; Immune response; Immune system; Immunophenotyping; Immunosuppressive Agents; In Situ; Inflammatory; Investigation; Knowledge; Lewy Body Dementia; Light; Link; Lymphocyte; Mediating; Microglia; Modeling; Molecular; Morphology; Mus; Myeloid Cells; Nature; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Outcome; Parkinson Disease; Parkinson' s Dementia; Pathogenesis; Pathologic; Pathology; Patients; Peripheral; Phenotype; Plant Roots; Play; Procedures; Proteins; Regulation; Research; Resolution; Risk; Role; Signal Transduction; Site; Spleen; Synapses; Syndrome; T-Lymphocyte; TREM2 gene; Technology; Testing; Therapeutic; Therapeutic Intervention; Time; Transgenic Mice; Transgenic Organisms; Treatment Efficacy; alpha synuclein; base; biomarker development; biomarker identification; cell type; cytokine; genetic variant; genomic signature; innovation; insight; lymph nodes; monocyte; mouse model; nervous system disorder; neurodegenerative dementia; neuroinflammation; neuropathology; novel; preclinical study; receptor; response; single-cell RNA sequencing; synucleinopathy; transcriptomics

Neurodegenerative disorders such as Alzheimer's disease (AD), Parkinson's disease with dementia (PDD) and dementia with Lewy bodies (DLB) have no cure, but all three share signs of α-synuclein (α-syn) pathology and of neuroinflammation. Although mounting evidence supports the notion that the immune cell response to neurodegeneration plays an active role in the pathogenesis of the above disorders, anti-inflammatory therapies have failed, thus far, to provide any beneficial effects in neurodegenerative diseases. We hypothesize that since immune cells exhibit a phenotypic heterogeneity, effective immune response-modifying therapy for neurodegenerative diseases requires the targeting of specific components of neuroinflammation rather than the broad inhibition of its signaling. The rationale for this project is that, once the regional and temporal genomic signatures of immune cells both inside and outside of the central nervous system (CNS) of animal models of synucleinopathies are known, meaningful biomarkers can be identified and innovative therapeutic strategies can be devised for PDD, DLB and even perhaps AD. Thus, the following two aims are proposed. To define the heterogeneity of the immune cell response to α-syn pathology within the CNS, in AIM 1, we will define brain immune cell subpopulations in transgenic (Tg) mThy1-α-syn mouse line 61 (a recognized model of synucleinopathies) by droplet-based single-cell RNA-sequencing (scRNASeq) using freshly extracted immune cells from different brain regions at time points ranging from 2 (no/minimal neuroinflammation) to 14 months (overt neuroinflammation); non-Tg littermates will be used as controls. Using computational techniques adapted to low-depth single-cell RNA-seq data, we will identify transcriptomic subtypes of immune cells from the different mouse brain regions; this includes estimates of cluster robustness, characterization of cell type-specific genes, and identification of marker genes. In AIM1, we will also validate the single-cell data by probing for sets of newly- defined and existing marker genes in situ. Since α-syn pathology also takes place outside of the CNS, in AIM 2, we will define the phenotypic diversity of the systemic immune cell response to α-syn pathology in blood, spleen and lymph nodes from Tg Line 61 (and non-Tg littermates) at the same selected time points as in AIM 1, and will be subjected to the same scRNA-seq procedure and cell type analyses. We will then develop novel bi- compartmental models to relate changes in CNS and peripheral immune system cells, with the goal of identifying putative interactions between these two sets of cells in synucleinopathies. Successful completion of the proposed investigations will establish an atlas of immune cell phenotype heterogeneity in synucleinopathies and shed light into the cross talk between immune cells localized inside and outside of the CNS in response to α-syn pathology. These findings will have an important positive impact on this area of research in that they will provide opportunities for biomarker identification and for therapeutic interventions and will advance our mechanistic understanding of PDD and DLB as well as possibly AD.

神经退行性疾病，例如阿尔茨海默病 (AD)、帕金森病伴痴呆 (PDD) 和 路易体痴呆 (DLB) 无法治愈，但这三种疾病都有 α-突触核蛋白 (α-syn) 病理学迹象，并且 尽管越来越多的证据支持免疫细胞对神经炎症做出反应的观点。 神经退行性变在上述疾病的发病机制中发挥着积极作用，抗炎治疗 迄今为止，尚未对神经退行性疾病产生任何有益效果。 免疫细胞表现出表型异质性，有效的免疫反应修改疗法 神经退行性疾病需要针对神经炎症的特定成分，而不是 该项目的基本原理是，一旦区域和时间基因组被广泛抑制。 动物模型中枢神经系统（CNS）内部和外部免疫细胞的特征 突触核蛋白病是已知的，可以识别有意义的生物标志物，并且可以采用创新的治疗策略 为 PDD、DLB 甚至 AD 设计。因此，提出以下两个目标来定义。 中枢神经系统内免疫细胞对 α-syn 病理学反应的异质性，在 AIM 1 中，我们将定义大脑 转基因 (Tg) mThy1-α-syn 小鼠 61 系（公认的免疫细胞模型）中的免疫细胞亚群 使用新鲜提取的免疫进行基于液滴的单细胞 RNA 测序 (scRNASeq) 来自不同大脑区域的细胞，时间点范围从 2 个月（无/轻微神经炎症）到 14 个月 (明显的神经炎症)；非Tg同窝动物将使用经过调整的计算技术作为对照。 根据低深度的单细胞 RNA-seq 数据，我们将识别来自不同来源的免疫细胞的转录组亚型 小鼠大脑区域；该估计包括簇稳健性、细胞类型特异性基因的表征， 在 AIM1 中，我们还将通过探测新的组来验证单细胞数据。 由于 α-syn 病理学也发生在 CNS 之外，在 AIM 2 中， 我们将定义全身免疫细胞对血液、脾脏中 α-syn 病理学反应的表型多样性 和来自 Tg 系 61（和非 Tg 同窝小鼠）的淋巴结，在与 AIM 1 相同的选定时间点，并将 然后我们将开发新的双-seq程序和细胞类型分析。 区室模型将中枢神经系统和外周免疫系统细胞的变化联系起来，目的是识别 突触核蛋白病中这两组细胞之间的假定相互作用成功完成。 拟议的研究将建立突触核蛋白病中免疫细胞表型异质性的图谱和 揭示了中枢神经系统内外免疫细胞之间对 α-syn 的反应 这些发现将对这一研究领域产生重要的积极影响，因为它们将提供 生物标志物识别和治疗干预的机会，并将推进我们的机制 了解 PDD 和 DLB 以及可能的 AD。