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

基本信息

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
项目类别：
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&apos 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&apos 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

Adult Alleles Alzheimer&apos 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&apos 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

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项目摘要

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，例如阿尔茨海默氏病（AD），帕金森氏病（PDD）和 Lewy身体（DLB）的痴呆症无法治愈，但所有三个共享症状的症状（α-Syn）病理学和神经炎症。尽管安装证据支持了免疫细胞反应的观念神经变性在上述疾病的发病机理中起积极作用，抗炎疗法到目前为止，未能在神经退行性疾病中提供任何有益作用。我们假设这是因为免疫细胞暴露了表型异质性，有效的免疫反应修改治疗治疗神经退行性疾病需要针对神经炎症的特定成分，而不是广泛抑制其信号传导。该项目的理由是，一旦区域和临时基因组免疫细胞的特征在中枢神经系统（CNS）的动物模型内外的特征突触核苷是已知的，可以确定有意义的生物标志物，创新的治疗策略可以设计为PDD，DLB甚至可能是AD。这是提出以下两个目标的。定义 IMNOCEL对CNS内的免疫球反应的异质性，在AIM 1中，我们将定义大脑转基因（TG）MTHY1-α-Syn小鼠系61中的免疫细胞亚群（公认的模型突触核酸）通过基于液滴的单细胞RNA序列（SCRNASEQ）使用新鲜提取的免疫时间点的不同大脑区域的细胞范围从2（无/最小神经炎症）到14个月（明显的神经炎症）；非TG同窝窝将用作控件。使用改编的计算技术对于低深度单细胞RNA-seq数据，我们将鉴定来自不同不同细胞的转录组亚型小鼠脑区域；这包括群集鲁棒性的估计，细胞类型特异性基因的表征，和标记基因的鉴定。在AIM1中，我们还将通过探测新的新数据来验证单细胞数据原位定义和现有标记基因。由于α-syn病理也发生在CNS之外，因此在AIM 2中我们将定义系统性免疫细胞对血液中α-Syn病理的反应的表型多样性，Sleen 与AIM 1相同的选定时间点，来自TG第61行（和非TG同窝式）的淋巴结和进行相同的SCRNA-SEQ程序和细胞类型分析。然后，我们将开发新颖的双隔室模型与中枢神经系统和外周免疫系统细胞的变化相关联，目的是识别这两组细胞在剧明病中的推定相互作用。成功完成拟议的调查将建立一个在剧明病中的免疫细胞表型异质性的地图集将光线放入CNS内部和外部的免疫电池之间的串扰，以响应α-syn 病理。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记识别和治疗干预措施的机会，并将提高我们的机械了解PDD和DLB以及可能的AD。