High-Throughput Analysis of Amyloid Deposition

淀粉样蛋白沉积的高通量分析

基本信息

批准号：
7092796
负责人：
CHAN B PARK
金额：
$ 2.03万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-05-01 至 2006-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7092796
关键词：
amyloid proteins biotechnology chemical aggregate drug screening /evaluation high throughput technology inhibitor /antagonist microfluidics molecular assembly /self assembly neuritic plaques stimulant /agonist technology /technique development

项目摘要

DESCRIPTION (provided by applicant): High-Throughput Analysis of Amyloid Deposition The formation of amyloid deposits in tissue is a hallmark of many pathologies such as Alzheimer's, Parkinson's, type II diabetes, mad cow, and other life-threatening diseases. Despite many studies on amyloid over a century, a clear understanding on how normal and soluble proteins assemble into fibrillar and insoluble amyloid aggregates under certain conditions remains unresolved. The key to unraveling the underlying etiology of amyloid-related diseases lies in the design of controlled in vitro experiments that can allow amyloid deposition study under selected environmental conditions that are known to induce normal protein to undergo aggregation. Amyloid deposition, the process of amyloid growth by the association of individual soluble amyloid molecules on natural amyloid template (i.e. plaque), is known to be critical for amyloid formation in vivo. To date, however, the use of 'natural' amyloid template to study amyloid deposition is extremely difficult and cumbersome. There is a critical need for high-throughput analytical system that enables a 'multi-factorial' investigation of a number of different environmental factors on amyloid deposition. The overall objective of this R21 proposal is to develop novel in vitro characterization methods that will lead to better understanding of amyloid deposition and to more effective screening of therapeutics. To meet the objective, four specific aims are proposed: (1) Design 'synthetic' templates suitable for amyloid deposition study in vitro, (2) Develop a microfluidic system for high-throughput analysis of amyloid deposition, (3) Validate amyloid deposition results obtained with microfluidic system, and (4) Conduct multi- factorial studies for screening inhibitors/accelerators of amyloid deposition/dissociation. The microfluidic system is intended to provide a novel way to study simultaneously how multiple environmental factors affect amyloid deposition/dissociation process while requiring only nano-liter amounts of amyloidogenic proteins and reagents. Results obtained from this research are expected to give deeper insight into the mechanism of amyloid deposition, as well as, to provide a powerful high-throughput protocol for screening therapeutics for amyloid-related diseases.

描述（由申请人提供）：淀粉样蛋白沉积的高通量分析组织中淀粉样蛋白沉积的形成是许多病理学的标志，例如阿尔茨海默病、帕金森病、II型糖尿病、疯牛病和其他危及生命的疾病。尽管一个多世纪以来对淀粉样蛋白进行了许多研究，但对正常和可溶性蛋白质如何在某些条件下组装成纤维状和不溶性淀粉样蛋白聚集体的清晰认识仍未解决。阐明淀粉样蛋白相关疾病的根本病因学的关键在于受控体外实验的设计，这些实验可以在已知会诱导正常蛋白质发生聚集的选定环境条件下进行淀粉样蛋白沉积研究。淀粉样蛋白沉积是通过单个可溶性淀粉样蛋白分子在天然淀粉样蛋白模板（即斑块）上结合而产生的淀粉样蛋白生长过程，已知对于体内淀粉样蛋白形成至关重要。然而，迄今为止，使用“天然”淀粉样蛋白模板来研究淀粉样蛋白沉积是极其困难和麻烦的。迫切需要高通量分析系统，能够对淀粉样蛋白沉积的许多不同环境因素进行“多因素”研究。 R21 提案的总体目标是开发新颖的体外表征方法，从而更好地了解淀粉样蛋白沉积并更有效地筛选治疗方法。为了实现这一目标，提出了四个具体目标：（1）设计适合体外淀粉样蛋白沉积研究的“合成”模板，（2）开发用于淀粉样蛋白沉积高通量分析的微流体系统，（3）验证淀粉样蛋白沉积结果用微流体系统获得，(4)进行多因素研究以筛选淀粉样蛋白沉积/解离的抑制剂/加速剂。该微流体系统旨在提供一种新的方法来同时研究多种环境因素如何影响淀粉样蛋白沉积/解离过程，同时仅需要纳升量的淀粉样蛋白和试剂。这项研究获得的结果有望更深入地了解淀粉样蛋白沉积的机制，并为筛选淀粉样蛋白相关疾病的治疗方法提供强大的高通量方案。