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

Pi：Yang，ong Proposal number：1227766本项目的目标是开发具有可调弹性的高清晰微纳米杂化拓扑学的组合库，以建立神经细胞淀粉样斑块形成的体外模型，以了解和缓解阿尔茨海默病(AD)。总体目标是描述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，并发现治疗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)的神经病理特征，而AD是导致老年人痴呆的最主要原因。它是由大脑中聚集的淀粉样β蛋白(A-β)多肽在细胞外沉积引起的。活体研究表明，淀粉样斑块的形成会改变局部微环境，随后导致神经元改变，最终导致神经退化。抑制淀粉样斑块的形成是治疗AD的一个可行的靶点。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识别治疗靶点的进展。因此，迫切需要以仿生的方式开发细胞培养技术，这种技术可以重复应用于调节神经细胞的行为，以促进AD研究和治疗开发所需的淀粉样斑块的形成。由于神经细胞和A-β聚集对底物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清晰微纳混合拓扑结构上培养神经细胞来重现淀粉样斑块的形成。为了验证这一假设，PI提出了以下具体任务：1)产生高度清晰的微纳杂化拓扑学；2)在纳米尺度上精确调整水凝胶的弹性水平；3)设计具有可调弹性的微纳杂化拓扑学组合库，以建立淀粉样斑块形成的体外细胞模型。该项目完成后，PI有望开发出必要的技术，以产生清晰的微纳杂化拓扑学，并在纳米尺度上微调基质弹性。在这些使能技术的支持下，构建了弹性可调的微纳混合拓扑库，以促进体外淀粉样斑块形成细胞模型的建立，这不仅对于阐明淀粉样斑块形成的基础至关重要，而且将有助于理解和缓解AD。更广泛的影响：作为这项提议不可或缺的一部分，计划开展广泛的教育和外联活动，以扩大任职人数不足群体的个人的参与。研究生和本科生，特别是女性和少数民族学生，将参与到多学科研究的方方面面。研究生将在西弗吉尼亚大学的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，并欣赏分子和细胞层面的细胞培养和表征技能。本科生将获得实践研究经验，以加深他们对科学原理的理解，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联活动，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的G9-12学生，特别是年轻女性和少数族裔，参加科学或工程领域的活动。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，以激发孩子们的兴奋，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。

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.