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

项目摘要/摘要 糖尿病在美国和世界范围内的患病率正在急剧上升，而新的药物 能够保护或恢复分泌胰岛素的胰岛β细胞的功能是迫切需要的。为 几十年来，这一领域的药物发现努力一直受到缺乏一致性和丰富性的阻碍 用于药物筛选的人β细胞。从诱导多能干细胞衍生的β细胞的出现 由再生医疗解决方案(RMS)开创的(IPSC)开启了一个随时可用的时代 β细胞是对化合物文库进行高通量筛选(HTS)的现实。相当可观 包括直接客户反馈在内的市场研究表明，HTS应用程序仍然昂贵 在对化合物文库进行筛选分析以检测胰岛素时也很费力，目前依赖于 基于抗体的方法，如免疫荧光显微镜和酶联免疫吸附 检测(ELISA法)。进一步提高筛选数千个糖尿病化合物文库的效率 在药物发现方面，RMS将设计用于HTS分析的iPSC生产线。这种经过改造的贝塔细胞产品将 在人胰岛素启动子控制下表达荧光素酶-绿色荧光蛋白融合蛋白。这 融合蛋白可以通过廉价和容易的自动化检测来检测，这将使IPSC衍生 在RMS的HTS产品平台中打包的Beta Cells，将成为一种真正准确、高效和经济的工具 高通量药物筛选应用。客户的直接反馈导致了此建议和解决方案 其执行HTS应用程序的特定实用程序要求。

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.