DISCO-MS: Proteomics of spatially identified tissues in whole organs and organisms

Spatial molecular profiling of complex tissues is essential to investigate cellular function in physiological and pathological states. However, methods for molecular analysis of biological specimens imaged in 3D as a whole are lacking. Here, we present DISCO-MS, a technology combining whole-organ/ism imaging, deep learning-based image analysis, robotic tissue extraction and ultra-high sensitivity mass spectrometry. DISCO-MS yielded qualitative and quantitative proteomics data indistinguishable from uncleared samples in both rodent and human tissues. Using DISCO-MS, we investigated microglia activation along axonal tracts after brain injury and characterized early and late-stage individual amyloid-beta plaques in Alzheimer's disease mouse model. Furthermore, aided by DISCO-bot robotic extraction we studied regional proteomics heterogeneity of immune cells in intact mouse bodies and aortic plaques in whole human heart. Overall, DISCO-MS enables unbiased proteome analysis of pre-clinical and clinical tissues after unbiased imaging of entire specimens in 3D, providing new diagnostic and therapeutic opportunities for complex diseases.

空间分子图谱技术在探究生理及病理状态下细胞功能方面至关重要。然而，对于三维整体成像的生物标本的分子分析方法尚显不足。在此，我们提出了一种名为DISCO-MS的技术，该技术融合了整器官/体成像、基于深度学习的图像分析、机器人组织提取以及超灵敏质谱分析。DISCO-MS技术产生的蛋白质组学数据在老鼠和人类组织中与未经处理的样本无显著差异，实现了定性和定量分析。利用DISCO-MS，我们研究了脑损伤后轴突通路上的小胶质细胞激活，并在阿尔茨海默病小鼠模型中描述了早期和晚期单个淀粉样斑块。此外，借助DISCO-bot机器人提取技术，我们还研究了完整小鼠体内的免疫细胞区域蛋白质组异质性和整个人心主动脉斑块。总之，DISCO-MS技术使得在三维无偏见成像整个标本之后，能够对临床前和临床组织进行无偏见的蛋白质组分析，为复杂疾病提供了新的诊断和治疗机会。