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：
10378725
负责人：
SERGE E PRZEDBORSKI
金额：
$ 44.59万
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/10378725
关键词：
ALS pathology 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 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 Tumor-infiltrating immune cells amyotrophic lateral sclerosis therapy 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 therapeutically effective 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.

肌萎缩性外侧硬化症（ALS）是一种致命运动神经元（MN）疾病，与神经炎症。尽管越来越多的证据支持神经炎症可能发挥活性的观念在ALS发病机理中的作用，抗炎疗法必须提供NO或最小的疾病修饰在ALS中的效果。本文中，我们假设由于免疫细胞表现出表型异质性，有效 ALS的免疫反应改良治疗需要靶向神经炎症的特定成分而不是广泛抑制其信号传导。这项研究的理由是，一旦基因组签名中枢神经系统中的免疫细胞（CNS）和ALS的周围神经系统（PNS）是可以确定已知的有意义的生物标志物，可以设计创新的治疗策略。因此，提出了以下三个目标。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执行单细胞RNA测序（SCRNASEQ） ALS和ALS患者的脊髓（ALS易感区域）和海马（ALS耐药区）从pre-pre- 有症状到末期瘫痪。然后，我们将在计算上使用此大规模多变量数据集构建与MN变性相关的集成CNS免疫细胞反应签名。自电机以来轴突变性是ALS病理学的关键特征，并发生在CNS外，在AIM 2中，我们将对周围神经浸润适应性和先天免疫细胞的表型分析在来自ALS患者和TG Mutsod1小鼠的坐骨神经中，使用与AIM 1相同的分析管道。然后，我们将在计算上构建一个双室模型，该模型整合了CNS和PNS免疫细胞信息以产生ALS的神经炎症签名。最后，由于脊髓的子区域在ALS中不均匀地退化，我们将使用AIM 1＆2中的集成的神经炎症签名产生至：（i）从ALS患者和TG询问现有的，全面的空间转录组数据库 mutsod1小鼠，通过免疫荧光和共聚焦显微镜（II）定位于不同的脊髓各自组织中的免疫细胞亚群。成功完成拟议的调查将响应于PN和CNS中ALS中免疫细胞表型的异质性神经变性。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病原体假设，用于鉴定生物标志物和用于ALS的治疗干预措施相关疾病。