The combinatorial effects of familial mutations, oxidative stress, and aging on BBB dysfunction in Alzheimer's disease

家族突变、氧化应激和衰老对阿尔茨海默病 BBB 功能障碍的综合影响

• 批准号: 10464625
• 负责人: Tracy D. Chung
• 金额: $ 4.68万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 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 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10464625
• 关键词: Address; Affect; Age; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease risk; American; Amyloid; Amyloid beta-Protein; Animals; Autopsy; Biological; Biological Assay; Blood; Blood - brain barrier anatomy; Blood Circulation; Blood Vessels; Categories; Cells; Cerebral Amyloid Angiopathy; Cerebrovascular system; Communication; Complex; Cues; Data; Data Set; Development; Disease; Disease Progression; Endothelial Cells; Etiology; Exposure to; Fellowship; Functional disorder; Future; Gene Expression; Gene set enrichment analysis; Goals; Human; Hydrogen Peroxide; Image; Immunohistochemistry; Impaired cognition; Impairment; Individual; Induced Mutation; Institutes; Institution; Intervention; Leadership; Libraries; Memory Loss; Mentorship; Microfabrication; Modeling; Molecular; Mutation; Nerve Degeneration; Neurofibrillary Tangles; Onset of illness; Outcome; Oxidative Stress; Pathologic; Pathology; Pathway interactions; Perfusion; Permeability; Pharmacology Study; Phenotype; Physiological; Prevention; Proteomics; Publishing; Research Training; Resolution; Role; Senile Plaques; Serum; Structural defect; Symptoms; Therapeutic; Tissue Engineering; Tissues; Toxin; Training; Transgenic Mice; Universities; Validation; Vascular Diseases; abeta deposition; age related; aged; base; blood-brain barrier disruption; blood-brain barrier function; brain cell; brain endothelial cell; brain tissue; career; combinatorial; design; disability; education resources; epidemiology study; experimental study; extracellular; familial Alzheimer disease; genetic signature; glucose transport; human old age (65+); imaging study; in vitro Model; in vivo; nanobiotechnology; occludin; presenilin-1; response; skills; solute; spatiotemporal; stem cell differentiation; stem cells; success; transcriptome sequencing; transcriptomics

Project Abstract Alzheimer’s disease is a disease of neurodegeneration and aging that affects millions of Americans, and is expected to impact millions more without further significant breakthroughs.1 However, much of the etiology and progression of Alzheimer’s is still unclear, especially given the complex interactions of many different molecular, cellular, and environmental cues that are correlated with phenotypic outcomes. An emerging focus in Alzheimer’s study is the role of the cerebrovasculature in the initiation, progression, and exacerbation of symptomatic disease.2-4 Disruption of the blood-brain barrier, which tightly controls any exchange between systemic circulation and brain tissue, has manifested in post-mortem and in vivo studies of late-stage Alzheimer’s disease as microbleeds, dysfunctional glucose transport, and impaired efflux of toxins;5 additional animal studies have indicated that some vascular dysfunction precedes neuronal degeneration in the progression of the disease.6, 7 Thus, to understand the drivers and progression of Alzheimer’s disease in hopes of identifying therapeutic breakpoints, the role of blood-brain barrier dysfunction must be investigated. To do so, we propose using a tissue-engineered model of the blood-brain barrier with high spatiotemporal resolution to assess its dysfunction under three key categories of perturbation associated with Alzheimer’s disease. These perturbations will span cell-intrinsic mutations associated with Alzheimer’s (APP(Swe) and PSEN(M146V)), extrinsic cues of oxidative stress (hydrogen peroxide exposure), and the systemic influence of aged blood components (exposure to aged vs. young human serum). We hypothesize that this combinatorial approach will best recapitulate human BBB phenotype in Alzheimer’s, and allow for modular study of each contributor. These perturbations will be compared transcriptomically, proteomically, and functionally. Transcriptomic changes will be studied through bulk RNA-sequencing and gene set enrichment analysis to highlight similarities to published human datasets, identify hallmark pathways that are impacted by Alzheimer’s cues, and motivate functional assay design for further validation in the tissue-engineered model. Proteomic and functional assessments include changes to barrier function, cell identity, and validation of pathways implicated by transcriptomic analysis. This study will provide a deeper understanding of the role of the blood- brain barrier in Alzheimer’s progression and emphasize candidate targets for future intervention. This project and related research training will be conducted under the guidance of Dr. Peter Searson at Johns Hopkins University and the Institute for Nanobiotechnology. Skills including functional assay design, stem cell differentiation, microfabrication, and RNA-sequencing analysis will be supported by the educational resources available within the institution. Additional goals of the fellowship training period will incorporate professional development for future career success, with an emphasis on communication, mentorship, and leadership skills.

项目摘要 阿尔茨海默病是一种影响数百万美国人的神经退行性疾病和衰老疾病， 如果没有进一步的重大突破，预计将影响数百万人。1 然而，大部分病因 阿尔茨海默病的进展仍不清楚，特别是考虑到许多不同因素之间复杂的相互作用 与表型结果相关的分子、细胞和环境线索。新兴焦点 阿尔茨海默氏症的研究重点是脑血管系统在阿尔茨海默氏症的发生、进展和恶化中的作用。 有症状的疾病。2-4 血脑屏障被破坏，血脑屏障严格控制着人与人之间的任何交换。 体循环和脑组织，已在后期的尸检和体内研究中得到体现 阿尔茨海默病表现为微出血、葡萄糖转运功能障碍和毒素流出受损；5 另外 动物研究表明，某些血管功能障碍先于神经元变性 6, 7 因此，要了解阿尔茨海默病的驱动因素和进展 为了确定治疗断点，必须研究血脑屏障功能障碍的作用。 为此，我们建议使用具有高时空特性的血脑屏障组织工程模型 决议评估与阿尔茨海默病相关的三个关键扰动类别下的功能障碍 疾病。这些扰动将跨越与阿尔茨海默氏症 (APP(Swe) 和 PSEN(M146V))、氧化应激的外在线索（过氧化氢暴露）以及系统性影响 老化的血液成分（暴露于老化与年轻的人类血清）。我们假设这个组合 该方法将最好地概括阿尔茨海默病中的人类 BBB 表型，并允许对每个 BBB 表型进行模块化研究 贡献者。这些扰动将从转录组、蛋白质组和功能上进行比较。 将通过批量 RNA 测序和基因集富集分析来研究转录组变化 突出与已发布的人类数据集的相似性，确定受阿尔茨海默氏症影响的标志性途径 线索，并激发功能测定设计，以在组织工程模型中进一步验证。蛋白质组学 功能评估包括屏障功能的变化、细胞特性和通路验证 转录组分析表明。这项研究将使人们更深入地了解血液的作用 阿尔茨海默病进展中的脑屏障，并强调未来干预的候选目标。 该项目及相关研究培训将在约翰斯大学Peter Searson博士的指导下进行 霍普金斯大学和纳米生物技术研究所。技能包括功能测定设计、干细胞 分化、微加工和 RNA 测序分析将得到教育资源的支持 机构内可用。奖学金培训期的其他目标将包括专业 未来职业成功的发展，重点是沟通、指导和领导技能。