Elucidating microvascular contributions to cognitive impairment at single-cell resolution

项目摘要/摘要 全球有超过5500万人患有痴呆症。人口老龄化，没有有效的治疗方法 预计到2050年，这一数字将增加近两倍。脑部小血管疾病(CSVD)可导致 占所有痴呆症的45%，约占中风的20%，出现在大多数阿尔茨海默病患者中。CSVD 源于大脑大约400英里的氧气和营养输送的小动脉，小动脉， 小静脉和毛细血管；被认为是导致认知障碍的最重要的血管因素 和痴呆症(VCID)。CSVD的病理，如动脉硬化和微梗塞，潜伏地发展为 扼杀脑血流，破坏血脑屏障，引发神经炎症。近期 单核测序技术的发展已经开始阐明基本的分子和 人类神经系统疾病背后的细胞过程。他们发现了有选择性的脆弱细胞 人群，跨疾病阶段的转录功能障碍，以及遗传风险变异对 告知新的研究领域和治疗目标。然而，单核研究大多失去了人脑。 血管细胞原因不明，使得我们对CSVD的认识相对滞后。为了应对这一挑战，我们 发明血管分离和核提取测序(VINE-SEQ)以有效捕获人脑 冰冻死后脑的血管细胞类型用于单核分析。在这里，我们将结合Vine-seq 用多模式单核RNA和ATAC测序生成了第一份~190万份血管图谱 早期、中期和晚期CSVD与年龄匹配的认知正常对照。样本来自一家 经特殊组织的191个死后额中回分水岭组织的CSVD集合，通过MRI和 神经束成像，来自宗教秩序研究和记忆与衰老项目(ROSMAP)。样本 没有表现出令人困惑的非血管神经退行性病变，并有丰富的人口统计学注释， 基因组、病理学和纵向认知数据，将分子读数与表型联系起来。因此，我们将 将Vine-seq与多模式单核RNA和ATAC测序(SnRNA/ATAC-seq)相结合 制作第一份CSVD血管图谱(A1)；揭示CSVD进展的分子特征(A2)；以及 确定CSVD遗传风险的原因机制(A3)。由于血管风险因素是可修改的，洞察力 这里揭示的可能是了解和最终治疗VCID和混合病理性痴呆的关键。

Project Summary/Abstract Dementia afflicts over 55 million people worldwide. With an aging population and no effective treatments available, this number is projected to nearly triple by 2050. Cerebral small vessel disease (CSVD) causes up to 45% of all dementia, accounts for ~20% of strokes, and appears in most Alzheimer’s disease patients. CSVD arises from pathologies of the brain’s ~400 miles of oxygen and nutrient-delivering small arteries, arterioles, venules, and capillaries; and is recognized as the most important vascular contributor to cognitive impairment and dementia (VCID). CSVD pathologies, such as arteriolosclerosis and microinfarcts, develop insidiously to strangulate cerebral blood flow, compromise the blood-brain barrier, and trigger neuroinflammation. Recent developments in single-nucleus sequencing technologies have begun elucidating fundamental molecular and cellular processes underlying human neurological disease. They have revealed selectively vulnerable cell populations, transcriptional dysfunction across disease stages, and the mechanisms of genetic risk variants to inform new research areas and therapeutic targets. Yet, single-nucleus studies have mostly lost human brain vascular cells unknown reasons, leaving our understanding of CSVD lagging. To address this challenge, we invented Vessel Isolation and Nuclei Extraction for Sequencing (VINE-seq) to efficiently capture human brain vascular cell types from frozen postmortem brains for single-nucleus profiling. Here, we will combine VINE-seq with multimodal single-nucleus RNA and ATAC sequencing to generate a first ~1.9 million vascular atlas of early, mid-, and late-stage CSVD alongside age-matched cognitively normal controls. Samples come from a specially organized CSVD set of 191 postmortem midfrontal gyrus watershed tissue, prioritized via MRI and nerve tractography, from the Religious Order Study and Memory and Aging Project (ROSMAP). Samples exhibit no confounding non-vascular neurodegenerative pathology and are richly annotated with demographic, genomic, pathologic, and longitudinal cognitive data to link molecular readouts to phenotypes. Thus, we will combine VINE-seq with multimodal single-nucleus RNA- and ATAC- sequencing (snRNA/ATAC-seq) to generate a first vascular atlas of CSVD (A1); reveal the molecular signatures of CSVD progression (A2); and determine the causal mechanisms of CSVD genetic risk (A3). As vascular risk factors are modifiable, insights revealed here may be critical to understanding and ultimately treating VCID and mixed pathology dementias.