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Engineered pancreatic endocrine cells that report beta cell toxicity for use in high throughput screening applications

Engineered pancreatic endocrine cells that report beta cell toxicity for use in high throughput screening applications
工程化胰腺内分泌细胞可报告 β 细胞毒性,用于高通量筛选应用
批准号:
9201795
负责人:
Erik J Forsberg
金额:
$21.74万
依托单位国家:
美国
项目类别:
财政年份:
2016
资助国家:
美国
项目状态:
已结题
起止时间:
2016-09-16 至 2017-08-31

项目摘要

项目成果

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中文摘要
翻译
项目总结/摘要 糖尿病在美国和世界范围内的患病率正在急剧增加, 能够保护或恢复胰岛β细胞分泌胰岛素的功能是迫切需要的。为 几十年来,药物发现的努力在这一领域一直受到阻碍,缺乏一致的和丰富的 人类β细胞用于药物筛选。来源于诱导多能干细胞的β细胞的出现 (iPSC),作为再生医疗解决方案(RMS)的先驱,已经开启了一个时代, β细胞是进行化合物文库的高通量筛选(HTS)的现实。相当大 包括直接客户反馈在内的市场研究表明,高温超导应用仍然很昂贵 并且在进行化合物文库的筛选测定以检测胰岛素时费力, 基于抗体的方法如免疫荧光显微镜和酶联免疫吸附 测定(ELISA)。为了进一步提高筛选数千种糖尿病化合物库的效率 在药物发现方面,RMS将设计用于HTS测定的iPSC系。这种工程化的β细胞产品将 在人胰岛素启动子的控制下表达胰蛋白酶-绿色荧光蛋白融合体。这 融合蛋白可以通过廉价且易于自动化的测定来检测, 在RMS HTS产品平台中封装的β细胞,是一种真正准确、高效和具有成本效益的工具, 高通量药物筛选应用。直接的客户反馈导致了这一建议和地址 他们执行HTS应用程序的特定实用程序要求。
英文摘要
Project Summary/Abstract The prevalence of diabetes in the United States and world-wide is increasing dramatically, and new drugs that can protect or restore the function of insulin-producing pancreatic beta cells are urgently needed. For decades, drug discovery efforts in this space have been hampered by the lack of consistent and abundant human beta cells for drug screening. The advent of beta cells derived from induced pluripotent stem cells (iPSC), as pioneered by Regenerative Medical Solutions (RMS), has opened an era of making readily available beta cells a reality for performing high-throughput screening (HTS) of compound libraries. Considerable market research, including direct customer feedback, has indicated that HTS applications remain expensive and laborious when conducting screening assays of compound libraries to detect insulin, which currently rely upon antibody-based methods such as immunofluorescence microscopy and enzyme-linked immunosorbent assays (ELISA). To further enhance the efficiency of screening thousands of compound libraries for diabetes drug discovery, RMS will engineer iPSC lines for use in HTS assays. Such engineered beta cell products will express a luciferase-green fluorescent protein fusion under the control of the human insulin promoter. This fusion protein, which can be detected by inexpensive and easily automated assays, will enable iPSC-derived beta cells, packaged in RMS’ HTS product platforms, to be a truly accurate, efficient, and cost-effective tool in high-throughput drug screening applications. Direct customer feedback has led to this proposal and addresses their specific utility requirements for performing HTS applications.
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会议论文
Development of a rapid and inexpensive luciferase-based high throughput screening assay to identify compounds that alter pancreatic β cell function
Diabetes drug screening platforms using patient pancreatic islet-like cells generated from induced pluripotent stem cells
Human Induced Pluripotent Stem Cell-Derived Beta-Cells for Drug and Toxicity Testing
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