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

基本信息

批准号：
9201795
负责人：
Erik J Forsberg
金额：
$ 21.74万
依托单位：
REGENERATIVE MEDICAL SOLUTIONS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-16 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9201795
关键词：
Address Amino Acids Antibodies Beta Cell Biological Assay Cell Count Cell Line Cell model Cells Chimeric Proteins Codon Nucleotides DNA DNA cassette Detection Development Diabetes Mellitus Drug Industry Drug toxicity Endocrine Engineering Enzyme-Linked Immunosorbent Assay Feedback Female Flow Cytometry Future Gene Expression Genes Genetic Engineering Genetic Transcription Glucose Green Fluorescent Proteins Human Immunofluorescence Immunologic Immunofluorescence Microscopy Insulin Karyotype Kinetics Libraries Luciferases Market Research Measures Medical Messenger RNA Methodology Methods Neomycin Pancreas Pharmaceutical Preparations Preclinical Drug Evaluation Prevalence Proteins Protocols documentation Reporter Reporter Genes Reporting Resistance Reverse Transcriptase Polymerase Chain Reaction Sampling Sequence Analysis Source Southern Blotting Staging Staining method Stains Structure of beta Cell of islet Testing Toxic effect Toxicity Tests Transfection Transgenes Transgenic Organisms United States abstracting base cost effective design diabetic patient drug discovery engineered beta cell flexibility functional restoration gene function high throughput screening induced pluripotent stem cell insulin secretion islet male meetings novel therapeutics pluripotency promoter regenerative response screening stem cell differentiation tool

项目摘要

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.

项目摘要/摘要在美国和世界范围内，糖尿病的患病率正在急剧增加，新药是迫切需要保护或恢复产生胰岛素的胰腺β细胞的功能。为了几十年来，由于缺乏一致和丰富的毒品发现工作受到阻碍人β细胞进行药物筛查。来自诱导多能干细胞的β细胞的冒险（IPSC），由再生医学解决方案（RMS）开创了一个易于使用的时代 Beta细胞是对化合物库进行高通量筛选（HTS）的现实。大量包括直接客户反馈在内的市场研究表明，HTS应用程序仍然很昂贵和实验室进行筛查测定法以检测胰岛素，目前依靠胰岛素基于抗体的方法，例如免疫荧光显微镜和酶联免疫吸附剂测定（ELISA）。为了进一步提高筛查数千个糖尿病的化合物库的效率 Drug Discovery，RMS将设计IPSC系列用于HTS测定法。这样的工程beta细胞产品将在人胰岛素启动子的控制下表达荧光素酶绿色荧光蛋白融合。这融合蛋白可以通过廉价且易于自动化的测定来检测到融合蛋白，将启用IPSC衍生的在RMS的HTS产品平台中包装的beta单元将成为真正准确，高效且具有成本效益的工具高通量药物筛查应用。直接客户反馈导致了此提案和地址他们执行HTS应用程序的特定效用要求。