High Resolution Profiling of Senescent Cells in ALS Brain and Spinal Cord

项目摘要/摘要 无论家族史、发病部位、症状和进展顺序如何，所有肌萎缩侧索硬化症 硬化症(ALS)患者失去肌肉质量，因为运动神经元连接到四肢肌肉进行控制 运动停止工作，最终消亡。然而，在最初的时间里，似乎有很大的延迟 神经元功能障碍和身体退化。复杂的神经系统之所以得以维持，部分原因是细胞 对压力的恢复能力，以维持功能并抵抗激活细胞死亡过程。在这方面的实验 提案将研究一种新的假设：ALS中的神经元作为复杂的 应激反应，避免主动退化和细胞死亡。然而，尽管这种保护机制可能 保护细胞的物理存在，衰老的细胞功能障碍和潜在的有毒分泌体促进 随着时间的推移，慢性组织退化，促进疾病的进展。蛋白质激活衰老 在其他神经退行性疾病中积聚。在这里我们将确定TDP43沉积 通过评估死后人脑和脊髓对细胞衰老应激反应的不同影响 来自资深捐赠者的脐带，通过退伍军人ALS生物仓库脑库访问，TDP43阳性 或阴性神经病理学。为了全面研究神经元及其环境，我们将利用 GeoMx数字空间剖面图技术。这种高分辨率、高含量的无损分析方法 在保持空间分辨率的同时，能够在单细胞水平上研究100种蛋白质。这 提供将数据映射回组织以将蛋白质表达差异与细胞相关联的能力 形态、在组织中的位置、细胞外环境以及与血管和/或其他病理的接近程度。 我们的研究提案产生的数据将为信息的深度提供前所未有的严谨性 肌萎缩侧索硬化症脊髓和脑的细胞应激。通过这些实验生成的丰富数据集将打开 许多研究ALS的新途径，并可能揭示细胞衰老作为一种新的治疗方法 目标。这是一个迫切需要发展的领域，因为大多数ALS患者平均死亡2-4人 在确诊数年后。

Project Summary/Abstract Regardless of family history, site of onset, and sequence of symptoms and progression, all Amyotrophic Lateral Sclerosis (ALS) patients lose muscle mass as the motor neurons that connect to muscle in limbs to control movement stop working and eventually die. However, there appears to be an extensive delay between initial neuronal dysfunction and physical degeneration. The complex nervous system is maintained, in part, due to cell resiliency to stress in order to maintain function and resist activating cell death processes. Experiments in this proposal will investigate a novel hypothesis: neurons in ALS enter cellular senescence as part of a complicated stress response to avoid active degeneration and cell death. Nonetheless, while this protective mechanism may preserve the physical presence of the cell, senescent cell dysfunction and potentially toxic secretome promotes chronic tissue degeneration over time that facilitates disease progression. Senescence is activated by protein accumulation in other neurodegenerative diseases. Here we will determine whether TDP43 deposition differentially affects the cellular senescence stress response by evaluating postmortem human brain and spinal cord from veteran donors, accessed through the VA ALS Biorepository Brain Bank, with either TDP43 positive or negative neuropathology. To comprehensively investigate neurons and their environment we will utilize GeoMx digital spatial profiling technology. This high resolution, high-content non-destructive analytical method enables the investigation of 100s of proteins at the single cell level while maintaining spatial resolution. This affords the ability to map the data back to the tissue to correlate protein expression differences with cell morphology, location in the tissue, extracellular environment and proximity to vascular and / or other pathologies. The data generated by our research proposal will provided an unprecedented rigor for depth of information on cellular stress in ALS spinal cord and brain. The rich dataset generated through these experiments will open many novel avenues of investigation in ALS and potentially unveil cellular senescence as a new therapeutic target. This is an area desperately in need of development as the majority of ALS patients die, on average, 2-4 years after diagnosis.