New technologies to identify molecular regulators of the human hippocampus neurogenic niche in healthy aging and Alzheimer's Disease

新技术识别健康老龄化和阿尔茨海默病中人类海马神经源性生态位的分子调节剂

• 批准号: 10620321
• 负责人: Maura Boldrini
• 金额: $ 80.58万
• 依托单位: NEW YORK STATE PSYCHIATRIC INSTITUTE DBA RESEARCH FOUNDATION FOR MENTAL HYGIENE, INC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-15 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10620321
• 关键词: Abeta synthesis; Adolescence; Adult; Affect; Age; Age Years; Aging; Alzheimer' s Disease; Amyloid beta-Protein; Anatomy; Animal Model; Animals; Anti-Inflammatory Agents; Autopsy; BETA2 protein; Bar Codes; Biological Assay; Birth; Brain; Brain region; Cell Differentiation process; Cell Nucleus; Cell Proliferation; Cell model; Cells; Cessation of life; Chromatin; Chromosome Mapping; Clinical; Collection; Custom; Data; Elderly; Electron Transport; Embryo; Enzyme-Linked Immunosorbent Assay; Epigenetic Process; Exclusion Criteria; Exons; Failure; Female; Gene Expression; Genes; Genetic Transcription; Genomics; Glial Fibrillary Acidic Protein; Hippocampus; Human; Immunofluorescence Immunologic; Impaired cognition; Impairment; Individual; Inflammation; Knowledge; Letters; Life; Location; Longevity; Maintenance; Mammals; Maps; Mass Spectrum Analysis; Messenger RNA; Metabolism; Methods; Mitochondria; Molecular; Molecular Target; Multipotent Stem Cells; Mus; NCAM1 gene; Neuroglia; Neuronal Plasticity; Neurons; Ontology; Population; Production; Proliferating; Proteins; Proteomics; RNA; RNA Splicing; Regional Blood Flow; Regulation; Reporting; Repression; Research; Resolution; Sampling; Slide; Small Nuclear RNA; Sudden Death; Synapses; Synaptic plasticity; Technology; Temporal Lobe; Testing; Time; Tissues; Toxicology; Transposase; Uniport; Vascular blood supply; Visualization; WNT Signaling Pathway; Western Blotting; adult neurogenesis; age related; angiogenesis; axon guidance; brain tissue; cell type; cognitive function; dentate gyrus; differential expression; emotional functioning; gliogenesis; healthy aging; indexing; juvenile animal; mRNA Expression; male; nerve stem cell; nestin protein; neurogenesis; neuropathology; new technology; normal aging; novel; polysialyl neural cell adhesion molecule; preservation; progenitor; protein expression; sex; single nucleus RNA-sequencing; stem; stem cell fate; stem cell proliferation; stem cells; synaptogenesis; tau Proteins; transcriptomics

The hippocampus neurogenic niche (HNN) generates new neurons in mammals, but it is unclear if this is happening in humans. Adult hippocampal neurogenesis is necessary to maintain intact cognitive and emotional functions regulated by the hippocampus. Markers of immaturity have been detected in neuronal cells of the HNN but it is still unclear if they represent adult-born neurons, or neuronal cells that have maintained their immaturity since birth. We found that the number of neural progenitor cells (NPCs) and immature neurons was stable throughout the eighth decade of life in normal aging (NA) subjects, but angiogenesis and neuroplasticity were decreased in older people. Other groups have supported our findings, while some could not detect immature neuronal cells in human hippocampus. Moreover, in aging mice, more NPCs differentiate into glia rather than neurons, compared to younger animals, but we do not know if this happens in humans. Adult neurogenesis is lower in Alzheimer’s Disease (AD) and it is unknown if this is because more NPCs differentiate into glia or through other mechanisms. These gaps in knowledge warrant the use of new technologies to investigate cellular lineages in the human HNN, and molecular regulators of NPCs proliferation, cell fate, differentiation, maturation and survival. This project aims to identify differentially expressed proteins (DEPs) and genes (DEGs) in the human HNN, at the regional and single cell level, comparing NA and AD. We will apply our pipeline using high resolution mass spectrometry for proteomics analysis, and single nuclei (sn) RNA and ATAC (Assay for Transposase- Accessible Chromatin) sequencing (seq), to identify gene expression and epigenetic changes. In slide-mounted hippocampus tissue, we will apply Visium (10X Genomics) and our custom-made spatial transcriptomic technology for anatomical co-mapping of cell-type specific mRNAs and proteins (DBiT-seq). Novel computational approaches will identify neurogenesis regulators in the human HNN that can be tested in cellular or animal models. Findings obtained with these “OMICS” approaches will be validated using HighPlex RNAscope® (ADCBio) and immunofluorescence, and qPCR, Western blots, and ELISA assays, to visualize and quantify DEP and DEG expression at the single cell and regional level. Our rigorous brain collection methods assure tissue quality, uniform processing, use of toxicology and neuropathology, and strict clinical inclusion/exclusion criteria. Groups include: NA subjects (N=100), Braak stage 0-1, age 14-99 yrs., 40 of which (60 years of age and older) are matched (by age, sex and postmortem interval between death and brain collection) with 40 AD cases (from the Columbia Taub Institute collection), Braak stage 1 through 4. Aims: 1. Identify HNN DEPs associated with NA and AD. 2. Identify DEGs in immature and mature neuronal and glial cell populations of the DG in NA and AD subjects, using sn-RNA and sn-ATAC-seq (10X Genomics). 3. Determine the anatomical localization of cell expressing DEGs and DEPs associated with NA and AD, using Visium and DBiT-seq. 4. Test correlations between DEPs and DEGs, and numbers of NPCs and immature neurons and glia in NA and AD.

海马神经源性生态位（HNN）在哺乳动物中产生新的神经元，但尚不清楚这是否是 发生在人类身上的海马神经发生对于维持完整的认知和情感是必要的。 在 HNN 神经元细胞中检测到了受海马体调节的功能。 但目前尚不清楚它们是否代表成年神经元，或保持不成熟的神经元细胞 我们发现自出生以来，神经祖细胞（NPC）和未成熟神经元的数量是稳定的。 在正常衰老（NA）受试者的整个生命的八个十年中，但血管生成和神经可塑性 其他团体也支持我们的发现，但有些团体无法检测出不成熟的人。 此外，在衰老小鼠中，更多的 NPC 分化为神经胶质细胞，而不是分化为神经胶质细胞。 与年轻动物相比，我们不知道这种情况是否会发生在人类身上。 阿尔茨海默氏病 (AD) 的比例较低，尚不清楚这是因为更多的 NPC 分化为神经胶质细胞还是通过 这些知识空白需要使用新技术来研究细胞谱系。 在人类 HNN 中，以及 NPC 增殖、细胞命运、分化、成熟和 该项目旨在识别人类中的差异表达蛋白（DEP）和基因（DEG）。 HNN，在区域和单细胞水平上，比较 ​​NA 和 AD，我们将使用高分辨率应用我们的流程。 用于蛋白质组学分析以及单核 (sn) RNA 和 ATAC（转座酶测定）的质谱分析 可访问染色质）测序 (seq)，用于识别载玻片中的基因表达和表观遗传变化。 海马组织，我们将应用 Visium (10X Genomics) 和我们定制的空间转录组 细胞类型特异性 mRNA 和蛋白质的解剖学联合绘图技术 (DBiT-seq)。 方法将鉴定人类 HNN 中的神经发生调节因子，并可在细胞或动物中进行测试 通过这些“组学”方法获得的模型将使用 HighPlex RNAscope® 进行验证。 (ADCBio) 和免疫荧光、qPCR、蛋白质印迹和 ELISA 测定，以可视化和量化 DEP 我们严格的大脑采集方法可确保组织在单细胞和区域水平上表达和表达 DEG。 质量、统一加工、毒理学和神经病理学的使用以及严格的临床纳入/排除标准。 分组包括：NA 受试者 (N=100)、Braak 0-1 期、年龄 14-99 岁，其中 40 名（60 岁及以上） 与 40 个 AD 病例（来自 哥伦比亚 Taub 研究所收藏），Braak 第 1 阶段到第 4 阶段。 目标： 1. 识别与以下相关的 HNN DEP： NA 和 AD 2. 识别 NA 和 AD 中 DG 的未成熟和成熟神经细胞和神经胶质细胞群中的 DEG。 AD 受试者，使用 sn-RNA 和 sn-ATAC-seq (10X Genomics) 3. 确定细胞的解剖定位。 使用 Visium 和 DBiT-seq 表达与 NA 和 AD 相关的 DEG 和 DEP 4. 测试相关性。 DEP 和 DEG 之间的关系，以及 NA 和 AD 中 NPC 和未成熟神经元和胶质细胞的数量。