对实验性自身免疫性脑脊髓炎中的中枢神经系统炎症、脱髓鞘和轴突损伤进行评分
February 23rd, 2024
实验性自身免疫性脑脊髓炎 （EAE） 是多发性硬化症的动物模型。本文介绍了一种对 EAE 中脊髓炎症、脱髓鞘和轴突损伤进行评分的方法。此外，还提出了一种量化小鼠血清中可溶性神经丝光水平的方法，有助于评估活小鼠的轴突损伤。
我们有兴趣了解自身免疫和多发性硬化症或多发性硬化症是如何开始的。本研究使用多种方法来研究风险因素如何与多发性硬化症相关。导致自身免疫的发展。实验性自身免疫性脑脊髓炎或 EAE 是 MS 的小鼠模型，可用于检查各种危险因素对自身免疫的影响。
我们最近发现了一种多发性硬化症危险因素肥胖如何导致小鼠自身免疫性炎症加剧的生物学途径。EAE模型的研究对于揭示这些机制至关重要。EAE 疾病严重程度通常按 5 分临床量表评分。
然而，该量表的敏感性有限，无法区分临床缺陷是由炎症还是组织损伤驱动的。脊髓和脑组织学是评估 EAE 炎症和组织损伤的金标准方法。该协议提供了一种简单的方法来测量EAE期间脊髓和大脑中的炎症和组织损伤。
我们的方案可以与临床量表一起使用，以更好地表征 EAE 的严重程度。我们的方案测量 EAE 的关键组织学特征，包括免疫浸润、脱髓鞘、脊髓轴突损伤和脑部炎症。它还包括测量小鼠血清中的神经丝光，这表明中枢神经系统损伤的程度。
与仅使用一种标记相比，该综合方案提供了更详细的EAE表征。
概述
本文介绍了一种用于评分实验性自身免疫性脑脊髓炎（EAE）模型中的脊髓炎症、脱髓鞘和轴突损伤的协议。它还详细说明了一种用于量化小鼠血清中可溶性神经丝蛋白轻链水平的方法，有助于评估轴突损伤。
关键研究组成部分
科学领域
- 神经科学
- 自身免疫
- 多发性硬化症研究
背景
- 实验性自身免疫性脑脊髓炎（EAE）是研究多发性硬化症的广泛使用的动物模型。
- 理解自身免疫的机制对于开发治疗策略至关重要。
- 当前的EAE评分方法在灵敏度和特异性方面存在局限性。
- 组织学分析对于评估炎症和组织损伤的程度至关重要。
研究目的
- 提供一个全面的协议，用于评估EAE中的脊髓和大脑病理。
- 在临床评分之外增强对EAE严重程度的表征。
- 测量神经丝蛋白轻链水平作为中枢神经系统损伤的生物标志物。
使用的方法
- 脊髓和大脑组织的组织学检查。
- 免疫浸润、脱髓鞘和轴突损伤的评分。
- 小鼠血清中神经丝蛋白轻链的定量。
- 组织收集和处理的详细手术技术。
主要结果
- 该协议允许对EAE病理进行详细评估。
- 组织学评分与临床疾病严重程度相关。
- 神经丝蛋白轻链水平提供了对轴突损伤的洞察。
- 在野生型和基因改造小鼠之间观察到了疾病严重程度的差异。
结论
- 该协议改善了对EAE病理的理解。
- 它提供了一种更灵敏的方法来评估疾病进展。
- 研究结果可能为未来的多发性硬化症治疗研究提供信息。
常见问题
Executive Industry Relevance
Quantitative scoring of central nervous system (CNS) inflammation, demyelination, and axonal injury in the EAE model addresses a critical gap in preclinical neuroinflammation research. Integrating histological and serum biomarker readouts enhances predictive confidence for target validation and mechanistic de-risking in multiple sclerosis (MS) drug discovery. This approach supports risk-adjusted portfolio decisions by providing translationally relevant endpoints beyond traditional clinical scoring.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables mechanistic interrogation of immune-mediated CNS injury pathways in MS models.
- Supports functional target validation by correlating genetic or pharmacological interventions with histological outcomes.
- Improves predictive confidence for advancing targets with disease-relevant CNS effects.
Screening & Assay Development
- Establishes validated histological and biomarker assays for quantifying inflammation, demyelination, and axonal injury.
- Facilitates reproducible, quantitative scoring systems for cross-study and cross-model comparisons.
- Enables robust screening of candidate interventions for CNS protection or repair.
Translational & Preclinical Research
- Aligns preclinical readouts with translational biomarkers such as serum neurofilament light (sNF-L).
- Provides continuity from discovery through preclinical validation by linking histological and functional outcomes.
- Supports risk-adjusted advancement of MS candidates based on mechanistic and biomarker evidence.
Pipeline & Workflow Integration
This protocol integrates into the discovery-to-preclinical continuum for neuroinflammatory and demyelinating disease programs.
- Discovery Biology: Enables hypothesis testing on immune cell infiltration, demyelination, and axonal injury mechanisms.
- Screening: Provides standardized, quantitative histological and biomarker assays for candidate evaluation.
- Analytics: Delivers reproducible scoring and quantitative outputs for cross-condition and cross-genotype comparisons.
- Translational Research: Bridges preclinical findings to clinical biomarker strategies using sNF-L measurements.
- Enterprise Reuse: Offers a reusable scoring and biomarker platform for diverse neuroinflammatory models.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence and reduces mechanistic ambiguity in CNS target validation.
- Operational Value: Standardizes histological and biomarker workflows for reproducibility and scalability.
- Strategic Value: Informs go/no-go decisions and reduces late-stage biological risk in MS portfolios.
- Portfolio Impact: Enables risk-adjusted prioritization and advancement of neuroinflammatory disease assets.
Implementation Considerations
- Requires expertise in neuroanatomy, histology, and quantitative image analysis.
- Needs access to tissue processing, immunostaining, and digital imaging infrastructure.
- Demands cross-team standardization of scoring criteria and biomarker assays.
- Adaptable to various rodent EAE models and potentially other neuroinflammatory systems.
- Dependent on rigorous sample handling and blinded analysis to ensure data integrity.
Biopharma FAQs
Chapters in this video
0:00
Introduction
1:48
Harvesting and Fixing Mouse Brain and Spinal Cord Tissues for Histological Scoring of Experimental Autoimmune Encephalomyelitis
3:15
Paraffin Embedding and Histopathological Scoring of Demyelinating Lesions and Perivascular Cuffs in Mouse Brain and Spinal Cord Tissues
