Multi-omic Brain Cell Atlas of Alzheimer's Disease Progression

项目总结/摘要 阿尔茨海默病（AD）是老年人中最常见的痴呆症，影响超过500万人 美国人，以及他们的家人和照顾者。不幸的是，全球人口老龄化只是 AD“流行病”恶化，因为预计到2050年发病率将增加两倍。尽管进行了深入的研究， 目前还无法治愈这种毁灭性的神经退行性疾病。因此，了解内在分子 驱动AD病理和进展的机制对于设计有效的治疗至关重要。死后 对人类大脑的检查显示，AD相关的神经病理学，如神经纤维缠结， （NFT）和神经变性，通常以保守的时空模式出现，影响经内嗅神经系统。 区域，然后延伸到边缘和等皮质区域。分子和神经化学基础 这种选择性神经元易损性（SNV）长期以来一直受到关注，因为它们是疾病进展的基础， 可能是理解神经退行性变的分子基础的关键，但到目前为止， 机制仍然难以捉摸。在这里，尖端的单细胞技术将用于产生一个 在不同阶段的AD易感脑区域内细胞类型的全面，多组学图谱 疾病该假设是，易感人群中受AD病理影响最严重的特定细胞类型 大脑区域的特征在于不同的分子通路（转录因子，信号级联，基因 网络）驱动SNV。此外，这些途径是在一个连续的时空模式执行， 通过染色质结构和基因调控元件的改变。追踪分子变化 通过这些神经元细胞群在AD病理连续体中的作用将更好地定义AD的发作和进展， 并可能指示新的治疗靶点。死后的大脑样本将从健康的 对照，或在死亡时表现出不同阶段的AD病理学的患者，即早期（Braak III/IV）或晚期（Braak III/IV）AD病理学的患者。 （Braak V/VI）。将分析在AD进展的不同阶段发生的变化以鉴定细胞类型。 以及对AD相关病理的起始和扩散最关键的分子途径。分析的区域将 海马（CA 1/sub）、下颞叶皮层（BA 20）、额叶皮层（BA 9）和视觉皮层（AD-1）。 抗性区）。在目标1中，将对对照受试者的样本进行单细胞分析，以表征 甲基化组和染色质结构联合（sn-m3 C-seq），以及染色质可及性， 转录组（Paired-seq），AD易感脑区域内的细胞类型。这些数据集的整合将 创建相关细胞类型的多组学图谱，作为了解AD发病的基础， 进展在目标2中，这些分析将扩展至AD患者。比较不同大脑区域的数据集 和疾病阶段将揭示AD中受影响最大的特定细胞类型，以及分子途径 （基于甲基化模式、染色质结构等的变化）在AD（Aim 3）中驱动SNV。

PROJECT SUMMARY/ABSTRACT Alzheimer's disease (AD) is the most common form of dementia in the elderly, affecting more than 5 million Americans, as well as their families and caregivers. Unfortunately, aging of the global population is only worsening the AD “epidemic”, as incidence is projected to triple by 2050. Despite intense research, there is currently no cure for this devastating neurodegenerative disorder. Thus, understanding the intrinsic molecular mechanisms that drive AD pathology and progression is critical to devising effective treatments. Postmortem examination of human brains has revealed that AD-associated neuropathologies, such as neurofibrillary tangles (NFTs) and neurodegeneration, generally arise in a conserved spatio-temporal pattern, affecting transentorhinal regions first, and later extending to limbic and isocortical areas. The molecular and neurochemical bases for such selective neuronal vulnerability (SNV) have long been pursued, as they underlie disease progression and may hold the key to understanding the molecular underpinnings of neurodegeneration, but to date these mechanisms remain elusive. Here, cutting-edge, single-cell technologies will be used to generate a comprehensive, multi-omic atlas of cell types within AD-vulnerable brain regions across different stages of disease. The hypothesis is that specific cell types most dramatically affected by AD pathology within susceptible brain regions are characterized by distinct molecular pathways (transcription factors, signaling cascades, gene networks) that drive SNV. Moreover, that these pathways are executed in a sequential spatio-temporal pattern by changes in chromatin architecture and gene regulatory elements. Tracking the molecular changes exhibited by these neuronal cell populations in the continuum of AD pathology will better define AD onset and progression, and potentially indicate new therapeutic targets. Postmortem brain samples will be obtained from healthy controls, or patients who at death exhibited different stages of AD pathology, namely early (Braak III/IV), or late (Braak V/VI). Changes occurring at different stages of AD progression will be analyzed to identify the cell types and molecular pathways most critical for the initiation and spread of AD-related pathology. Regions analyzed will be hippocampus (CA1/sub), inferior temporal cortex (BA20), frontal cortex (BA9), and visual cortex (an AD- resistant region). In Aim 1, samples from control subject will be subjected to single cell analyses to characterize the methylome and chromatin architecture jointly (sn-m3C-seq), as well as chromatin accessibility together with transcriptome (Paired-seq), of cell types within AD-vulnerable brain regions. Integration of these datasets will create a multi-omic atlas of relevant cell types that will serve as the foundation for understanding AD onset and progression. In Aim 2, these analyses will be extended to AD patients. Comparing data sets across brain regions and disease stages will reveal the specific cell types most affected in AD, as well as the molecular pathways (based on changes in methylation patterns, chromatin architecture, etc.) that drive SNV in AD (Aim3).