Development of novel genomic approaches for profiling cellular temporal-spatial dynamics of neurogenesis in Aging and Alzheimer's disease

项目总结 成人神经发生正在成为维持大脑动态平衡和正常功能的重要参与者。 神经发生功能障碍与衰老和神经紊乱有关，包括 阿尔茨海默病(AD)。系统地绘制神经发生的分子动力学图的能力 细胞分辨率可以作为系统工作的基础，以便更好地理解 在衰老和疾病中引起细胞状态异常。虽然单细胞基因组学的快速发展是 创造了前所未有的机会来探索哺乳动物大脑中的分子异质性，几乎所有这样的 这些方法被限制在低吞吐量，无法从根本上恢复数据的异构性和动态性 成人神经发生中的罕见细胞状态(例如，不到大脑中细胞总数的0.1%)。在此，我们建议 开发新的方法学，全面了解神经发生的时空动态 在衰老和阿尔茨海默病(AD)期间，人类和小鼠的大脑都会受到影响。具体地说，我们将首先开发一个 一种新的高通量、低成本的单细胞基因组学方法，SciNext1000，用于分析分子 人死后海马区样本中400万个细胞的异质性。这种方法将是强大的 因为我们不仅可以定量地表征人类成年海马区神经发生的频率 单细胞分辨率，但也识别与衰老中受损的神经发生相关的转录组特征 异构体分辨率下的AD。此外，我们还将开发另一种新的单细胞基因组技术-SCI-Div-Dv。 SEQ，以加强对新生神经元的检测，并确定细胞分化轨迹和 幼年和老年小鼠脑中成年神经发生的相关转录特征。生成的数据集 将促进我们对不同神经谱系和神经发生中的基因调控的理解 构成了向全面描述潜在的分子机制迈出的重要一步 衰老过程中的神经发生功能障碍。除了内部分子程序，神经发生过程是 由来自神经干龛的环境信号控制。我们将应用高吞吐量 空间转录策略识别成人参与的细胞相互作用和局部微环境 人类和小鼠大脑中的神经发生。这些多管齐下的方法将开启一种新的范式 了解成人神经发生的全球分子程序和环境调控，从而 提供潜在的治疗靶点，以恢复衰老和大脑疾病中的细胞种群动态平衡。

PROJECT SUMMARY Adult neurogenesis is emerging as an important player in maintaining brain homeostasis and normal functions. The dysfunctions of neurogenesis have been associated with aging and neurological disorders, including Alzheimer’s disease (AD). The ability to systematically map the molecular dynamics of neurogenesis at single- cell resolution could serve as a foundation for a systematic effort to better understand the molecular events that give rise to abnormal cell states in aging and diseases. While the rapid advances in single-cell genomics are creating unprecedented opportunities to explore molecular heterogeneity in mammalian brains, nearly all such methods are restricted to low throughput and fail to recover the heterogeneity and dynamics of the profoundly rare cell states in adult neurogenesis (e.g., less than 0.1% of the cell population in the brain). Herein, we propose to develop novel methodologies that enable a comprehensive view of temporal-spatial dynamics of neurogenesis during aging and Alzheimer's disease (AD) in both human and mouse brains. Specifically, we will first develop a novel high-throughput, low-cost single-cell genomics approach, sciNext1000, to profile the molecular heterogeneity of four million cells from post-mortem human hippocampal samples. This approach will be powerful because we can not only quantitatively characterize the frequency of human adult hippocampal neurogenesis at single-cell resolution, but also identify the transcriptome features associated with impaired neurogenesis in aging and AD at isoform resolution. In addition, we will develop another novel single-cell genomic technique, sci-Div- seq, to enhance the detection of newborn neurons, and identify the cellular differentiation trajectories and associated transcriptomic features of adult neurogenesis in young and aged mouse brains. The resulting dataset will advance our understanding of gene regulation in neurogenesis across different neural lineages and constitute a significant step towards a comprehensive characterization of the molecular mechanism underlying neurogenesis impairment in aging. In addition to the internal molecular programs, the neurogenesis process is controlled by aspects of environmental signals from the neural stem niche. We will apply a high-throughput spatial transcriptomic strategy to identify the cellular interactions and local microenvironment involved in adult neurogenesis in both human and mouse brains. These multi-pronged approaches will open a new paradigm for understanding the global molecular programs and environmental regulation of adult neurogenesis, thereby informing potential therapeutic targets to restore cell population homeostasis in aging and brain disorders.