Moloney Christina M, Rutledge Matthew H, Labuzan Sydney A, Peng Zhongwei, Tranovich Jessica F, Wood Ashley C, Rothberg Darren M, Duara Ranjan, Lachner Christian, Graff-Radford Neill R, Dickson Dennis W, Kanaan Nicholas M, Carter Rickey E, Murray Melissa E
Department of Neuroscience, Mayo Clinic, Jacksonville, FL, 32224, USA.
Department of Quantitative Health Sciences, Mayo Clinic, Jacksonville, FL, 32224, USA.
bioRxiv. 2025 Jun 6:2025.06.02.657435. doi: 10.1101/2025.06.02.657435.
Neurofibrillary tangles are dynamic neuropathologic hallmarks of Alzheimer's disease with a hypothesized lifespan morphologically-defined by three maturity levels: pretangles, mature tangles, and ghost tangles. To better understand the progression of tangle pathophysiology, we characterized tangle maturity level predilection of 15 tau antibodies recognizing a broad range of linear, phosphorylation, conformational, and truncation epitopes in the hippocampus of 24 postmortem brains. We developed the tangle maturity scoring system to semi-quantitatively evaluate each tangle maturity level. Based on proportions of tangle maturity levels, we classified antibodies as "early" (mostly pretangles and mature tangles), "middling" (mature tangles with pretangles and ghost tangles), and "advanced" (mostly ghost tangles and mature tangles) tangle maturity markers. To summarize tangle maturity predilection, we developed the tangle maturity scale to integrate individual tangle maturity scores. Correlations showed stronger relationships between tangle maturity scale and subsector thickness for more advanced tangle maturity markers in CA1 and subiculum, whereas Braak tangle stage remained consistently correlated throughout markers of the tangle lifespan. To aid in scoring hippocampi, we used machine learning to recognize tangle maturity levels, which performed comparably to a domain expert and showed similar relationships by Spearman correlation. Pattern recognition software was used to assess tangle and neuritic tau burden separately, which generally correlated with Braak stage and neuronal counts. However, tangle-derived tau burden more consistently correlated with hippocampal subsector thickness. In conclusion, we developed manual and automated scoring systems to evaluate tangle maturity levels, demonstrating early 4R, phosphorylated, and oligomeric tau accumulation preceding more advanced 3R and truncated tau. Our study provides supportive evidence of disease-relevant ordering of tau posttranslational modifications in the brain, which may have implications for theragnostic development. These findings underscore the promise of computerized quantitative analyses (i.e., pathomics) for high-throughput feature extraction from whole-slide images to enhance our understanding of microscopically observed morphologic changes.
神经原纤维缠结是阿尔茨海默病的动态神经病理学特征，其假定寿命在形态学上由三个成熟水平定义：前缠结、成熟缠结和幽灵缠结。为了更好地理解缠结病理生理学的进展，我们对15种tau抗体的缠结成熟水平偏好进行了表征，这些抗体识别24个死后大脑海马体中广泛的线性、磷酸化、构象和截短表位。我们开发了缠结成熟度评分系统，以半定量评估每个缠结成熟水平。基于缠结成熟水平的比例，我们将抗体分类为“早期”（主要是前缠结和成熟缠结）、“中期”（成熟缠结与前缠结和幽灵缠结）和“晚期”（主要是幽灵缠结和成熟缠结）缠结成熟度标志物。为了总结缠结成熟度偏好，我们开发了缠结成熟度量表，以整合个体缠结成熟度评分。相关性分析表明，对于CA1和海马下托中更晚期的缠结成熟度标志物，缠结成熟度量表与亚区厚度之间的关系更强，而Braak缠结阶段在缠结寿命的所有标志物中始终保持相关性。为了辅助海马体评分，我们使用机器学习来识别缠结成熟水平，其表现与领域专家相当，并且通过Spearman相关性显示出相似的关系。模式识别软件用于分别评估缠结和神经突tau负荷，其通常与Braak阶段和神经元计数相关。然而，源自缠结的tau负荷与海马亚区厚度的相关性更一致。总之，我们开发了手动和自动评分系统来评估缠结成熟水平，证明在更晚期的3R和截短tau之前，早期4R、磷酸化和寡聚tau会积累。我们的研究为大脑中tau翻译后修饰的疾病相关顺序提供了支持性证据，这可能对治疗诊断学的发展具有启示意义。这些发现强调了计算机化定量分析（即病理组学）从全切片图像中进行高通量特征提取以增强我们对微观观察到的形态学变化理解的前景。
