Development of in vivo quantitative stable isotope probing to quantify microbiome dynamics in Alzheimers Disease

开发体内定量稳定同位素探测以量化阿尔茨海默病微生物组动态

• 批准号: 10301904
• 负责人: Emily K Cope
• 金额: $ 22.8万
• 依托单位: NORTHERN ARIZONA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10301904
• 关键词: 16S ribosomal RNA sequencing; 3xTg-AD mouse; AD transgenic mice; Address; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease risk; Amyloid; Amyloid beta-Protein; Amyloidosis; Bacteria; Blood - brain barrier anatomy; Brain; Cardiovascular system; Cessation of life; Communities; DNA; DNA sequencing; Data; Death Rate; Dementia; Density Gradient Centrifugation; Deposition; Development; Disease Progression; Ecology; Ecosystem; Elderly; Enteric Nervous System; Etiology; Exhibits; Gastrointestinal tract structure; Goals; Growth; Health; Human; Human Microbiome; Human body; Immune; Immune system; Individual; Inflammatory; Isotope Labeling; Isotopes; Label; Longitudinal Studies; Measures; Methods; Microbe; Mouse Strains; Mus; Nature; Nerve Degeneration; Neurofibrillary Tangles; Neurologic; Neurotransmitters; Nucleic Acids; Pathogenesis; Pathologic; Pathology; Patients; Phase III Clinical Trials; Population; Population Dynamics; Process; Production; Proteins; Quality of life; RNA; Research; Research Personnel; Role; Senile Plaques; Severities; Shotguns; Soil; Structure; Tauopathies; Taxon; Techniques; Technology; Time; Transgenic Organisms; Vagus nerve structure; Volatile Fatty Acids; Water; Wild Type Mouse; abeta accumulation; aging population; base; beta pleated sheet; blood-brain barrier crossing; cognitive function; density; disorder prevention; disorder risk; gut microbiome; gut microbiota; gut-brain axis; host microbiome; host-associated microbial communities; human disease; human microbiota; in vivo; innovation; insight; interest; member; metagenomic sequencing; microbial; microbial community; microbial host; microbiome; microbiome composition; microbiome research; microbiome sequencing; microbiota; microorganism; mouse model; neuroinflammation; neuropathology; new technology; novel; pathogenic microbe; stable isotope; tool; vagus nerve stimulation; β-amyloid burden

ABSTRACT Alzheimer’s Disease (AD) is the leading cause of dementia in the aging population and severely impacts patient’s and caretaker’s quality of life. Despite decades of research, the underlying etiology of AD is incompletely understood. The prevailing hypothesis for AD pathology is the amyloid cascade hypothesis, which posits that the causative agent of AD is the accumulation of amyloid-β (Aβ, the main component of plaques) in the brain. However, recent Phase III clinical trials that target Aβ have not led to significant improvements in cognitive function, despite reducing overall brain Aβ burden. There has been a growing interest in understanding additional pathological features of AD that may impact disease progression. Recent studies in humans and in mice have suggested a role for pathogenic microbes or altered microbiota in neuroinflammation and AD. Microbiota in the GI tract can influence neurological health through microbial production of neurotransmitter precursors, immune-modulatory metabolites (e.g. short chain fatty acids), or interaction with the vagus nerve or enteric nervous system. Gut microbiota can also produce amyloids (aggregated, insoluble proteins exhibiting β-pleated sheet structures) that may cross the blood-brain barrier. Despite growing efforts to understand the gut microbiome-brain axis, current technologies using DNA or RNA amplicon sequencing are unable to address fundamental ecological questions, such as quantifying taxon-specific growth rates of host- associated microbiota. Understanding microbial ecosystem dynamics has large implications for human disease; quantifying growth or turnover of species in terms of absolute abundance change over time in an ecosystem can enable predictions of interspecies competition or trajectories of microbial succession in early and late life, which are associated with host health or disease risk. In this study, we will adapt an innovative technique widely used in soil microbial ecology to study host-microbiome dynamics in AD. We will use quantitative stable isotope probing (qSIP), a technique that uses an isotopically enriched substrate (e.g. 18O-water) to measure gut microbiome dynamics in triple transgenic (3xTg-AD) and wild- type mice. Since H2O is a universal substrate, labeled 18O will incorporate into microbial and host biomolecules, including nucleic acids; this feature allows a researcher to separate extracted DNA by density to quantify taxon-specific growth rates. Completion of this study will lead to the development of a novel tool that can be widely used in the AD community to better understand the contribution of host- associated microbiota to AD progression and neuroinflammation.

抽象的 阿尔茨海默氏病（AD）是衰老人口痴呆的主要原因，严重 影响患者和看守人的生活质量。尽管进行了数十年的研究，但 广告不完全理解。 AD病理学的主要假设是淀粉样蛋白级联 假设认为AD的病因是淀粉样蛋白β的积累（Aβ，主要 大脑中斑块的成分。但是，靶向Aβ的最近的III期临床试验尚未引导 为了显着改善认知功能，目的地减轻了整体大脑Aβ负担。有 人们对理解AD的其他病理特征的兴趣越来越大，可能会影响 疾病进展。在人类和小鼠中的最新研究提出了致​​病性的作用 神经炎症和AD中的微生物或微生物群改变。胃肠道中的微生物群会影响 通过微生物产生神经递质前体，免疫调节的神经健康健康 代谢产物（例如短链脂肪酸）或与迷走神经或肠神经系统的相互作用。 肠道菌群还可以产生淀粉样蛋白（骨料，不溶性的蛋白 结构）可能越过血脑屏障。尽管越来越多地了解肠道 微生物组 - 脑轴，使用DNA或RNA扩增子测序的当前技术无法 解决了基本的生态问题，例如量化宿主的分类单元特异性增长率 - 相关的微生物群。了解微生物生态系统动力学对人具有很大的影响 疾病;根据绝对抽象随着时间的推移而量化物种的生长或营业额 生态系统可以启用种间竞争的预测或微生物成功的轨迹 在早期和晚期，与宿主健康或疾病风险有关。在这项研究中，我们将适应 一种在土壤微生物生态学中广泛用于研究AD中宿主微生物组动力学的创新技术。 我们将使用定量稳定的同位素探测（QSIP），该技术使用同位素富集 底物（例如18o水）测量三重转基因（3xtg-ad）和野生的肠道微生物组动力学 类型的鼠标。由于H2O是通用的底物，因此标记的18O将纳入微生物和宿主 生物分子，包括核酸；此功能使研究人员可以通过 量化分类群特异性增长率的密度。这项研究的完成将导致 可以在广告社区中广泛使用的新颖工具，以更好地了解宿主的贡献 - 与AD进展和神经炎症相关的菌群。