Endogenous barcoding to determine complex dynamics of adult neurogenesis in aging and Alzheimer's disease

内源条形码确定衰老和阿尔茨海默病中成人神经发生的复杂动态

• 批准号: 10434404
• 负责人: GRIGORI N ENIKOLOPOV
• 金额: $ 80.96万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10434404
• 关键词: Acceleration; Address; Adult; Adverse effects; Aging; Alleles; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Amyloid beta-Protein; Animal Model; Animals; Bar Codes; Birth; Brain; Brain Diseases; Cell Differentiation process; Cells; Clonal Expansion; Clonality; Cognitive; Complement; Complex; DNA; Development; Disease; Drug usage; Excitatory Amino Acid Antagonists; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic Recombination; Goals; Hippocampus (Brain); Human; Impaired cognition; Impairment; Individual; Knowledge; Label; Levetiracetam; Life Cycle Stages; Link; Longevity; Maintenance; Malignant - descriptor; Memantine; Methods; Modality; Modeling; Monoclonal Antibodies; Mus; Neurons; Outcome; Pharmaceutical Preparations; Physical activity; Production; RNA; Regulation; Reporter; Resolution; Running; Scheme; Series; System; Techniques; Tissues; adult neurogenesis; aging brain; cognitive function; cohort; combinatorial; human old age (65+); instrument; mature animal; mouse genome; mouse model; nerve stem cell; neurodevelopment; neurogenesis; normal aging; novel strategies; single cell analysis; stem cell division; stem cells; tool; transcriptome; transcriptomics

ABSTRACT New neurons in the adult human and animal hippocampus have been implicated in several cognitive functions. These functions are profoundly impaired by the loss or insufficient production of new neurons. Neurons are produced after a prolonged series of transitions including the activation, proliferation, elimination, and differentiation of neural stem cells and their progeny. Competing hypotheses have been proposed to describe how these steps are executed. These hypotheses imply divergent, sometimes contradicting, blueprints of the transitions from stem cells to neurons, as well as different outcomes when these blueprints are perturbed by aging or disease. Alzheimer's disease (AD) has a dramatic adverse effect on hippocampal neurogenesis in humans and in animal models, and this decrease is thought to be directly linked to the cognitive dysfunction observed in AD. Identifying the changes in neurogenesis induced by AD and differentiating them from the changes induced by aging may reveal new means of mitigating or even reversing AD-associated cognitive impartment. This goal is challenging not only because of the complexity of the system, but also because of the limitations inherent in currently available approaches for tracing stem cells and their progeny. We have recently developed a novel approach of endogenous barcoding to determine the lineage trajectory and differentiation trajectories of neural stem cells in the adult mouse brain. In addition, we have developed a new technique for combinatorial multitag labeling of subpopulations of dividing stem cells and their progeny. Now we propose applying these new approaches to identify the changes that AD and aging induce in the basic scheme of neural stem cell division and differentiation and to examine how those changes can be mitigated. In our first specific aim, we will introduce the Polylox barcode cassette and related genes into the genome of mouse models of AD, apply recombination-induced endogenous barcoding, and perform barcode analysis integrated with single-cell transcriptome analysis as a novel approach for determining the division, differentiation, and lineages of individual neural stem cells, as well as the changes inflicted by AD and aging. In our second aim, we will apply our new technique of multitag labeling of dividing cells to further determine the dynamics of adult neurogenesis and perturbations introduced by AD and aging. Together, these approaches will provide a new means of dissecting the perturbations in neurogenesis provoked by aging and AD. Finally, in our third aim, we will determine how these changes are modified by drugs used in different therapy modalities, using them here as experimental instruments for further revealing the AD- and aging-induced changes.

抽象的 成年人和动物海马体中的新神经元与多种认知功能有关 功能。这些功能因新神经元的丧失或产生不足而受到严重损害。神经元 经过一系列长期的转变后产生，包括激活、增殖、消除和 神经干细胞及其后代的分化。已经提出了竞争性假设来描述 这些步骤是如何执行的。这些假设意味着不同的、有时甚至是矛盾的蓝图 从干细胞到神经元的转变，以及当这些蓝图受到干扰时的不同结果 衰老或疾病。阿尔茨海默病（AD）对海马神经发生有显着的不利影响 人类和动物模型中，这种下降被认为与认知功能障碍直接相关 在 AD 中观察到。识别 AD 引起的神经发生变化并将其与 AD 区分开来 衰老引起的变化可能揭示减轻甚至逆转 AD 相关认知的新方法 传授。这个目标具有挑战性，不仅因为系统的复杂性，还因为 目前追踪干细胞及其后代的方法存在固有的局限性。 我们最近开发了一种内源条形码的新方法来确定谱系轨迹 以及成年小鼠大脑中神经干细胞的分化轨迹。此外，我们还开发了一个 对分裂干细胞及其后代进行组合多标签标记的新技术。现在 我们建议应用这些新方法来识别 AD 和衰老在基本功能中引起的变化 神经干细胞分裂和分化的方案，并检查如何减轻这些变化。 在我们的第一个具体目标中，我们将把 Polylox 条形码盒和相关基因引入基因组中 AD 小鼠模型，应用重组诱导的内源条形码，并进行条形码分析 与单细胞转录组分析相结合，作为确定分裂、分化、 个体神经干细胞的谱系，以及 AD 和衰老造成的变化。在我们的第二个 目标，我们将应用我们的分裂细胞多标签标记新技术来进一步确定分裂细胞的动态 AD 和衰老引起的成人神经发生和扰动。这些方法共同将提供 剖析衰老和 AD 引起的神经发生扰动的新方法。最后，在我们的第三个目标中， 我们将确定不同治疗方式中使用的药物如何改变这些变化，并使用它们 此处作为进一步揭示 AD 和衰老引起的变化的实验工具。