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

项目摘要 阿尔茨海默病是一种神经退行性疾病和衰老，影响数百万美国人， 在没有进一步重大突破的情况下，预计将影响数百万人。 阿尔茨海默氏症的发展仍然不清楚，特别是考虑到许多不同的疾病之间的复杂相互作用， 与表型结果相关的分子、细胞和环境线索。一个新的焦点 在阿尔茨海默氏症的研究中，血管系统在阿尔茨海默氏症的发生、发展和恶化中的作用是最重要的。 2 -4血脑屏障的破坏，血脑屏障严格控制着大脑与大脑之间的任何交换。 体循环和脑组织，表现在死后和体内研究的后期阶段 阿尔茨海默病作为微出血、葡萄糖转运功能障碍和毒素外排受损;5额外 动物研究已经表明，某些血管功能障碍先于神经元变性， 因此，为了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，我们建议使用一个组织工程模型的血脑屏障与高时空 在与阿尔茨海默氏症相关的三个关键扰动类别下， 疾病这些扰动将跨越与阿尔茨海默病相关的细胞内在突变（APP（Swe）和 PSEN（M146 V）），氧化应激的外在线索（过氧化氢暴露），以及 老化血液成分（暴露于老化与年轻人血清）。我们假设这种组合 这种方法将最好地概括阿尔茨海默氏症中的人类BBB表型，并允许对每个表型进行模块化研究。 贡献者。这些扰动将比较转录组，蛋白质组，和功能。 将通过批量RNA测序和基因集富集分析研究转录组学变化， 强调与已发表的人类数据集的相似性，确定受阿尔茨海默病影响的标志性途径 提示，并激发功能测定设计，用于在组织工程模型中进一步验证。蛋白组 功能评估包括屏障功能的变化、细胞特性和途径的验证 与转录组学分析有关这项研究将提供一个更深入的了解血液的作用- 脑屏障在阿尔茨海默病的进展，并强调候选目标，为未来的干预。 这个项目和相关的研究培训将在约翰大学彼得·瑟森博士的指导下进行 霍普金斯大学和纳米生物技术研究所。技能包括功能分析设计、干细胞 分化，微加工和RNA测序分析将得到教育资源的支持 在机构内可用。研究金培训期的其他目标将包括专业人员、 未来职业成功的发展，重点是沟通，指导和领导技能。

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.