High-Throughput Screen For FDA Approved Drugs That Amplify Beta-Cell Mass In Vivo

高通量筛选 FDA 批准的体内放大 β 细胞质量的药物

• 批准号: 8045193
• 负责人: Mehboob A Hussain
• 金额: $ 252.44万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8045193
• 关键词: Address; Adolescent; Animal Model; Automation; Beta Cell; Biological Models; Blood Glucose; Cell Cycle; Cell Cycle Progression; Cell Differentiation process; Cell Proliferation; Cells; Cellular biology; Chemicals; Chronic; Clinic; Collaborations; Collection; Detection; Diabetes Mellitus; Disease; Drug Compounding; Drug effect disorder; FDA approved; Fertility; Fishes; Genetic; Glucose; Hand; Human; Insulin; Insulin Resistance; Insulin-Dependent Diabetes Mellitus; Investigation; Islets of Langerhans; Knowledge; Larva; Libraries; Mammals; Methods; Modeling; Molecular; Monitor; Mus; Nature; Non-Insulin-Dependent Diabetes Mellitus; Pancreas; Pathology; Pathway interactions; Pharmaceutical Preparations; Preclinical Drug Evaluation; Process; Production; Proteins; Publications; Publishing; Relative (related person); Reporter; Request for Applications; Research; Research Personnel; Resources; Robotics; Route; Screening procedure; Site; Stem cells; System; Techniques; Technology; Testing; Therapeutic; Therapeutic Agents; Therapeutic Clinical Trial; Therapeutic Effect; Therapeutic Uses; Transgenic Organisms; Work; Zebrafish; abstracting; base; diabetes mellitus therapy; drug testing; experience; follow-up; high throughput screening; human disease; in vivo; innovation; interest; islet; mouse model; novel therapeutics; pancreas development; progenitor; skills; tool

DESCRIPTION (provided by applicant): High-throughput screen for FDA approved drugs that amplify Beta-cell mass in vivo The aim of the project proposed here is to identify drugs that will be therapeutic in the treatment of Diabetes. Type I (or Juvenile) Diabetes is caused by a chronic loss of the insulin-producing ¿-cells of the pancreas. Without insulin the body fails to regulate blood glucose levels leading to adverse pathology. There is a high degree of conservation in the molecular mechanisms that control vertebrate pancreas development; therefore, we have turned to the zebrafish as an alternative model system to study ¿-cell biology. As zebrafish larvae are small, and fit inside single wells of multiwell plates, they are the only vertebrate model system suited for carrying out high-throughput chemical screens. One of the largest problems in finding new therapeutic drugs is the huge lag between discovery and getting the drug into the clinic. To expedite this process, we use two approaches: 1) drugs are tested directly in animal models, and 2) the compound library we are screening - the Johns Hopkins Drug Library (JHDL) - is a unique resource of mainly FDA approved drugs. Thus, regardless of current use, any FDA drug found to be therapeutic in our research will have a significantly faster route into the clinic, and human trials. Our published work has shown that ¿-cell formation in the secondary islets of fish pancreata is functionally equivalent to ¿-cell neogenesis in mammals. Hence, drugs that cause early differentiation of ¿-cells in fish larvae are hypothesized to also induce ¿-cell neogenesis in mammals. Using transgenic zebrafish we performed a drug screen for inducers of precocious ¿-cell formation. From 3347 drugs evaluated we identified 5 'Hits'. As part of this proposal, we plan to validate these 'Hit' compounds in murine models of Type I diabetes. Furthermore, we intend to reinvestigate the JHDL to search for compounds that function at concentrations not tested in our initial screen, and to test promising leads in synergistic combinations. Finally, we will develop the tools for a second screen. This follow-up screen will again use transgenic zebrafish but will be aimed at identifying compounds that induce proliferation of endogenous ¿-cells. Between our 2 screens we aim to find compounds that induce ¿-cell formation and increase their continued production. In this way we hope to find FDA approved drugs that will increase ¿-cell mass in humans and that will be therapeutic in the treatment of diabetes. We also intend to use our knowledge of ¿-cell biology to elucidate the mode of action of the drugs we identify. This will further our understanding of ¿-cell neogenesis and reveal molecular pathways that could be exploited to develop better therapies. The methods and technologies we have developed for our screening are universal in nature, being based on relative expression levels of reporter proteins. Thus, these techniques will be of considerable interest to researchers investigating a wide array of human diseases. PUBLIC HEALTH RELEVANCE: High-throughput screen for FDA approved drugs that amplify Beta-cell mass in vivo In an effort to find therapies for diabetes, we have just finished an innovative high-throughput screen on larval zebrafish. We screened a library of FDA approved drugs for compounds that cause differentiation of ¿-cells. By validating the drugs most effective at inducing ¿-cell formation from our initial screen and drugs that induce proliferation in our proposed second screen, ultimately we aim to find drugs that can increase ¿-cell mass in humans.

描述（由申请人提供）：FDA批准的在体内扩增β细胞质量的药物的高通量筛选这里提出的项目的目的是确定将用于治疗糖尿病的药物。I型（或青少年）糖尿病是由胰腺分泌胰岛素的细胞的慢性损失引起的。没有胰岛素，身体就无法调节血糖水平，导致不良病理。控制脊椎动物胰腺发育的分子机制具有高度的保守性；因此，我们将斑马鱼作为研究细胞生物学的替代模型系统。由于斑马鱼幼体很小，适合在单孔或多孔板中生长，因此它们是唯一适合进行高通量化学筛选的脊椎动物模型系统。寻找新的治疗药物的最大问题之一是从发现到将药物投入临床之间的巨大滞后。为了加快这一过程，我们采用了两种方法：1)药物直接在动物模型中进行测试；2)我们筛选的化合物库——约翰霍普金斯药物库（JHDL）——主要是FDA批准的药物的独特资源。因此，无论目前的用途如何，在我们的研究中，任何FDA发现的治疗药物都将有一个明显更快的途径进入临床和人体试验。我们发表的研究表明，鱼类胰腺次级胰岛的细胞形成在功能上等同于哺乳动物的细胞新生。因此，在鱼类幼虫中引起细胞早期分化的药物被假设也能诱导哺乳动物的细胞新生。利用转基因斑马鱼，我们对早熟细胞形成的诱导剂进行了药物筛选。从评估的3347种药物中，我们确定了5种“命中”。作为该提案的一部分，我们计划在1型糖尿病的小鼠模型中验证这些Hit化合物。此外，我们打算重新研究JHDL，以寻找在初始筛选中未测试的浓度下起作用的化合物，并测试协同组合中有希望的线索。最后，我们将为第二个屏幕开发工具。后续的筛选将再次使用转基因斑马鱼，但目的是鉴定诱导内源性细胞增殖的化合物。在我们的两个筛选中，我们的目标是找到能够诱导细胞形成并增加其持续生产的化合物。通过这种方式，我们希望找到FDA批准的药物，可以增加人体细胞质量，并用于治疗糖尿病。我们还打算利用我们的细胞生物学知识来阐明我们鉴定的药物的作用方式。这将进一步加深我们对细胞新生的理解，并揭示可用于开发更好治疗方法的分子途径。我们开发的筛选方法和技术在本质上是通用的，基于报告蛋白的相对表达水平。因此，这些技术将对研究各种人类疾病的研究人员产生相当大的兴趣。