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

BRIGE：使用弹性可调的微纳米混合拓扑结构组合文库的淀粉样斑块形成的体外细胞模型

• 批准号: 1227766
• 负责人: Yong Yang
• 金额: $ 17.5万
• 依托单位: West Virginia University Research Corporation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1227766&HistoricalAwards=false
• 关键词: BRIGE; Vitro; Cellular; Model; Amyloid

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.

PI：Yang, Yong提案编号：1227766该项目的目标是开发具有可调弹性的高度定义的微纳米混合拓扑结构的组合库，以建立神经细胞淀粉样斑块形成的体外模型，以了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，并发现针对 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，而阿尔茨海默病是老年人痴呆的最主要原因。它是由大脑中聚集的淀粉样β (A-β) 肽的细胞外沉积引起的。体内研究表明，淀粉样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的淀粉样斑块的体外模型，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，可重复地应用于调节神经细胞行为，以促进淀粉样斑块的形成，用于 AD 研究和治疗开发。 由于神经细胞以及 A-β 聚集对基质的表面特性高度敏感，因此推测淀粉样斑块的形成可以通过在弹性可调的高度限定的微纳米混合拓扑上培养神经细胞来重现。为了检验这一假设，提出了以下具体任务的PI：1）产生高度定义的微纳米混合形貌； 2）在纳米尺度上精确调节水凝胶的弹性水平； 3) 设计具有可调节弹性的微纳米混合拓扑结构组合库，以建立淀粉样斑块形成的体外细胞模型。在该项目完成时，PI预计将开发出生产明确的微纳米所需的技术。纳米混合形貌并在纳米尺度上微调基底弹性。借助这些使能技术，可以设计具有可调弹性的微/纳米混合拓扑结构组合库，以促进淀粉样斑块形成的体外细胞模型的建立，这不仅对于阐明淀粉样斑块形成的基本原理至关重要，而且也将有助于理解和缓解AD。更广泛的影响：作为该提案不可或缺的一部分，计划开展广泛的教育和外展活动，以扩大代表性不足群体的个人参与。研究生和本科生，特别是女性和少数民族学生将参与多学科研究的各个方面。研究生将在西弗吉尼亚大学工程和健康科学校区工作，学习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，以加深对科学原理的理解，并将课堂知识与他们在现实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，并将其纳入西弗吉尼亚大学现有的外展活动，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探索博物馆的纳米日进行宣传，以激发孩子们的兴趣并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合作研究，并促进与当地行业的合作。