Defining immune cell heterogeneity in human ALS and mouse model of the disease

定义人类 ALS 和该疾病小鼠模型中的免疫细胞异质性

基本信息

批准号：
10034251
负责人：
SERGE E PRZEDBORSKI
金额：
$ 64.14万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-15 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10034251
关键词：
ALS patients Adult Amyotrophic Lateral Sclerosis Anatomy Anti-Inflammatory Agents Autopsy Axon Biological Markers Cells Characteristics Chronic Confocal Microscopy Cost of Illness Data Data Set Databases Disease Disease model Environment Exhibits Future Goals Heterogeneity Hippocampus (Brain)Human Immune Immune response Immune system Immunofluorescence Microscopy Immunosuppressive Agents In Situ Inflammatory Response Investigation Knowledge Maps Microglia Modeling Molecular Motor Motor Neuron Disease Motor Neurons Mus Myeloid Cells Nerve Degeneration Neuraxis Neurodegenerative Disorders Neuromuscular Junction Paralysed Pathogenesis Pathogenicity Pathology Patients Peripheral Nerves Peripheral Nervous System Phenotype Play Process Research Resistance Resolution Role Sampling Signal Transduction Spinal Cord Surface Technology Testing Therapeutic Therapeutic Intervention Time Tissues Transgenic Organisms Treatment Efficacy Tumor-infiltrating immune cells axon injury axonal degeneration base biomarker development biomarker identification genomic signature immunological diversity innovation macrophage monocyte motor neuron degeneration mouse model mutant nervous system disorder neuroinflammation novel response sciatic nerve single-cell RNA sequencing superoxide dismutase 1 transcriptome

项目摘要

Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron (MN) disease that is associated with features of neuroinflammation. Although mounting evidence supports the notion that neuroinflammation may play an active role in ALS pathogenesis, anti-inflammatory therapies have to provide either no or minimal disease-modifying effect in ALS. Herein, we hypothesize that since immune cells exhibit a phenotypic heterogeneity, effective immune response-modifying therapy for ALS requires the targeting of specific components of neuroinflammation rather than broadly inhibiting its signaling. The rationale for this research is that, once the genomic signatures of immune cells in the central nervous system (CNS) and the peripheral nervous system (PNS) of ALS are known, meaningful biomarkers can be identified and innovative therapeutic strategies can be devised. Thus, the following three aims are proposed. To define the heterogeneity of the immune cell response within the CNS, in AIM 1, we will perform single-cell RNA-sequencing (scRNASeq) using freshly extracted immune cells from spinal cord (ALS susceptible region) and hippocampus (ALS resistant region) of patients with ALS as well as of the extensively used and validated transgenic (Tg) mutant SOD1 (mutSOD1) mouse model of ALS, from pre- symptomatic to end-stage paralysis. We will then utilize this large-scale multivariable dataset computationally to construct an integrated CNS immune cell response signature associated with MN degeneration. Since motor axon degeneration is a critical feature of ALS pathology and takes place outside of the CNS, in AIM 2, we will perform a phenotypical analysis of peripheral nerve infiltrating adaptive and innate immune cells by scRNASeq in sciatic nerves from both ALS patients and Tg mutSOD1 mice using the same analytic pipeline as in AIM 1. We will then computationally construct a bi-compartmental model that integrates the CNS and PNS immune cell information to generate a neuroinflammatory signature of ALS. Lastly, since subregions of the spinal cord degenerate unevenly in ALS, we will use the integrated, neuroinflammatory signatures generates in AIM 1 & 2 to: (i) interrogate an existing, comprehensive spatial transcriptome database from both ALS patients and Tg mutSOD1 mice, and by immunofluorescence and confocal microscopy (ii) localize the different spinal cord immune cell subpopulations in the respective tissue. Successful completion of the proposed investigations will establish heterogeneity of the immune cell phenotype in ALS in both the PNS and CNS in response to neurodegeneration. These findings will have an important positive impact in that they will provide opportunities for novel pathogenic hypothesis, for identification of biomarkers and for therapeutic interventions in ALS and related disorders.

肌萎缩侧索硬化症 (ALS) 是一种致命的运动神经元 (MN) 疾病，与以下特征相关：神经炎症。尽管越来越多的证据支持神经炎症可能发挥积极作用的观点由于抗炎治疗在 ALS 发病机制中的作用，因此必须不提供或最小程度地改善疾病对 ALS 的影响。在此，我们假设，由于免疫细胞表现出表型异质性，有效的 ALS 的免疫反应调节疗法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。这项研究的基本原理是，一旦基因组特征 ALS 的中枢神经系统 (CNS) 和周围神经系统 (PNS) 中的免疫细胞可以识别已知的、有意义的生物标志物，并可以设计出创新的治疗策略。因此，提出以下三个目标。为了定义中枢神经系统内免疫细胞反应的异质性，在 AIM 1 中，我们将使用新鲜提取的免疫细胞进行单细胞 RNA 测序 (scRNASeq) ALS 患者的脊髓（ALS 易感区）和海马（ALS 抵抗区）以及广泛使用和验证的转基因 (Tg) 突变体 SOD1 (mutSOD1) 小鼠 ALS 模型有症状直至终末期瘫痪。然后我们将在计算上利用这个大规模多变量数据集构建与 MN 变性相关的集成 CNS 免疫细胞反应特征。自电机轴突变性是 ALS 病理学的一个关键特征，发生在 CNS 之外，在 AIM 2 中，我们将通过 scRNASeq 对周围神经浸润适应性和先天免疫细胞进行表型分析使用与 AIM 1 相同的分析流程，对 ALS 患者和 Tg mutSOD1 小鼠的坐骨神经进行分析。然后，我们将通过计算构建一个整合 CNS 和 PNS 免疫细胞的双室模型生成 ALS 神经炎症特征的信息。最后，由于脊髓的亚区域 ALS 中退化不均匀，我们将使用 AIM 1 和 2 中生成的集成神经炎症特征 (i) 查询来自 ALS 患者和 Tg 的现有综合空间转录组数据库 mutSOD1 小鼠，并通过免疫荧光和共聚焦显微镜 (ii) 定位不同的脊髓相应组织中的免疫细胞亚群。成功完成拟议的调查将建立 ALS 中 PNS 和 CNS 免疫细胞表型的异质性，以响应神经变性。这些发现将产生重要的积极影响，因为它们将提供机会用于新的致病假说、生物标志物的鉴定以及 ALS 和 ALS 的治疗干预相关疾病。