BRIGE: In Vitro Cellular Model of Amyloid Plaque Formation Using Combinatorial Libraries of Micro-nano-hybrid Topographies with Tunable Elasticity

主要研究者：杨勇提案编号：1227766该项目的目的是开发具有可调弹性的高度定义的微纳米混合形貌的组合库，以建立从神经细胞形成淀粉样蛋白斑块的体外模型，用于理解和减轻阿尔茨海默病（AD）。总体目标是描绘转基因神经细胞在仿生微环境中产生淀粉样斑块的分子机制，并发现抗AD的药物靶点。智力优势：淀粉样斑块的形成是阿尔茨海默病（AD）的神经病理学标志，AD是老年痴呆症的最主要原因。它是由聚集的淀粉样β（A-β）肽在脑中的细胞外沉积引起的。体内研究表明，淀粉样蛋白斑块的形成改变了局部微环境，随后导致神经元改变，最终导致神经退行性变。抑制淀粉样蛋白斑块形成代表了针对AD的可行治疗靶点。目前缺乏神经细胞形成淀粉样斑块的体外模型，这极大地限制了寻找抑制淀粉样斑块形成的策略和确定治疗靶点的进展。因此，迫切需要以仿生方式开发细胞培养技术，其可以可重复地应用于调节神经细胞行为以促进淀粉样蛋白斑块的形成，用于AD研究和治疗开发。 由于神经细胞以及A-β聚集体对基质的表面性质高度敏感，因此假设可以通过在具有可调弹性的高度限定的微纳米混合形貌上培养神经细胞来重现淀粉样蛋白斑块的形成。为了验证这一假设，PI提出了以下具体任务：1）产生高度限定的微纳米混合形貌：2）精确调节纳米级水凝胶的弹性水平; 3）设计具有可调弹性的微纳米混合形貌的组合库，以建立淀粉样蛋白斑块形成的体外细胞模型。PI期望已经开发出必要的技术来产生良好定义的微纳米混合形貌，并在纳米尺度上微调衬底弹性。利用这些技术，构建了弹性可调的微/纳混合形貌的组合库，有助于建立淀粉样斑块形成的体外细胞模型，这不仅对阐明淀粉样斑块形成的基础至关重要，而且有助于理解和缓解AD。更广泛的影响：作为这项建议不可或缺的一部分，已计划开展广泛的教育和外联活动，以扩大代表性不足群体的个人的参与。研究生和本科生，特别是妇女和少数民族学生将参与多学科研究的各个方面。研究生将在WVU的工程和健康科学校区工作，将教授聚合物纳米科学和纳米工程，并欣赏分子和细胞水平的细胞培养和表征技能。本科生将获得实践研究经验，加深他们对科学原理的理解，并将课堂知识与他们在真实的世界中观察到的现象联系起来。此外，将开发包括教学模块和实验室演示的项目，并将其整合到西弗吉尼亚大学现有的外展活动中，特别是为期一周的“明日工程师”夏令营，旨在鼓励阿巴拉契亚地区G9-12学生，特别是年轻女性和少数族裔，参与科学或工程领域。纳米生物技术将在西弗吉尼亚州儿童发现博物馆的纳米日进行宣传，以激发儿童的兴趣，提高公众对纳米生物技术的认识。该项目将进一步与健康科学研究人员进行合作研究，并促进与当地产业的合作。

PI: Yang, YongProposal Number: 1227766The objective of this project is to develop combinatorial libraries of highly defined micro-nanohybrid topographies with tunable elasticity to build an in vitro model of amyloid plaque formation from neural cells for understanding and alleviating Alzheimer's disease (AD). The overall goal is to delineate the molecular mechanism by which transgenic neural cells generate amyloid plaques in a biomimetic microenvironment and discover pharmaceutical targets against AD. Intellectual Merit: Amyloid plaque formation is a neuropathological hallmark of Alzheimer's disease (AD) which is the most prominent cause of dementia in the elderly. It is caused by the extracellular deposition of aggregated amyloid-beta (A-beta) peptides in the brain. In vivo studies reveal that the amyloid plaque's formation changes the local microenvironment, subsequently leading to neuronal alterations, eventually neurodegeneration. Inhibition of amyloid plaque formation represents a feasible therapeutic target against AD. The current lack of in vitro models of amyloid plaques formed from neural cells greatly limits the progress in seeking strategies to inhibit amyloid plaque formationand identifying therapeutic targets. There is, therefore, a critical need for the development of cell culture technologies in a biomimetic manner that can be reproducibly applied to regulate neural cell behavior to facilitate formation of amyloid plaques for AD research and therapeutic development. Because neural cells as well as A-beta aggregation are highly sensitive to surface properties of the substrate, it is hypothesized that the amyloid plaque formation can be recapitulated by culturing neural cells on highly defined micro-nano-hybrid topographies with tunable elasticity. To test this hypothesis, the PI following specific tasks are proposed: 1) Produce highly defined micro-nano-hybrid topography; 2) Precisely tune the level of elasticity of hydrogels at nanoscale; 3) Engineer combinatorial libraries of micro-nano-hybrid topographies with tunable elasticity to build an in vitro cellular model of amyloid plaque formation.At the completion of this project, the PI expects to have developed the technologies necessary to produce well-defined micro-nano-hybrid topography and to fine tune substrate elasticity at the nanoscale. With these enabling technologies, combinatorial libraries of micro-/nano hybrid topographies with tunable elasticity is engineered to facilitate the establishment of in vitro cellular model of amyloid plaque formation, which is not only crucial to the elucidation of the fundamental of amyloid plaque formation, but will also contribute to understand and alleviate AD. Broader Impacts: As an indispensable part of this proposal, extensive Education and Outreach activities have been planned to broaden the participation of individuals from underrepresented groups. Graduate and undergraduate students, especially women and minority students will be involved into every aspect of the multidisciplinary research. Graduate students will work on bothEngineering and Health Science campuses at WVU, will be taught polymer nanoscience and nanoengineering, and appreciate cell culture and characterization skills at the molecular and cellular levels. Undergraduate students will obtain hands-on research experience to deepen their understanding of scientific principles and to relate classroom knowledge to phenomena they observe in the real world. Moreover, projects comprising teaching modules and lab demonstrations will be developed and integrated into existing outreach events at WVU, especially the weeklong Engineers of Tomorrow summer camp designed to encourage Appalachian area G9-12 students, in particular young women and minorities, to participate in a science or engineering field. Nanobiotechnology will be publicized on Nano Days in Children's Discovery Museum of West Virginia to excite the children and increase public awareness of nanobiotechnology. This project will further collaborative research with health science researchers and foster collaborations with local industries.