The Development of a High-Throughput Assay to Screen Chemical Compounds in Human Pluripotent Stem Cell Derived Neural Cells

开发高通量检测方法来筛选人多能干细胞衍生神经细胞中的化合物

• 批准号: 9016471
• 负责人: Nathan C Boles
• 金额: $ 22.17万
• 依托单位: ATHGHIN BIOTECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9016471
• 关键词: Affect; Biological Assay; Biological Testing; Brain; Cell Line; Cerebral cortex; Cerebrum; Chemical Exposure; Chemicals; Chronic; Commerce; Data; Development; Developmental Gene; Exposure to; Gene Expression; Health; Human; Human Development; In Vitro; Industry; Laboratories; Lead; Malignant Neoplasms; Nature; Neurons; Pluripotent Stem Cells; Population; Process; Production; Protocols documentation; Psyche structure; Research Personnel; Staging; System; Testing; Time; Toxic effect; United States; autism spectrum disorder; consumer product; cost effective; disability; high throughput screening; human embryonic stem cell; induced pluripotent stem cell; novel; public health relevance; safety testing; stem cell technology; tool; transcriptomics

 DESCRIPTION (provided by applicant): The long-term objective of this application is to provide a high-throughput platform to test the effects of chemical exposure on human brain development. In the US, chemicals are not required for health or environmental safety testing before their incorporation in consumer products. Over 80,000 registered chemicals are in commerce, most of which are used without any information on their long-term health effects. Considering each one of these one by one is a slow and expensive process. Currently there is no system in place to screen compounds in terms of their health consequences before product implementation and with over 80,000 registered chemicals in commerce, a high-throughput mechanism to screen these compounds is needed. Human pluripotent stem cell (hPSC) technology offers a manipulatable platform to study the early stages of brain development. In a recent study from our lab, we adapted a protocol to differentiate hPSCs into cerebral cortical neurons over 77 days that recapitulated each stage of cortical development. The cerebral cortex has been shown to be particularly sensitive to chemical exposure during development. Preliminary data from our lab show that chemical exposure during the early stages of this protocol lead to changes of key neuro- developmental genes at different times during development providing evidence that our in vitro system can be used to assess chemical toxicity. Here we propose the development of a high-throughput assay using hPSC technology to screen large numbers of chemical compounds by assessing how they alter human brain development. This study will develop a cost-effective tool to analyze thousands of compounds with a simple PCR assay that can be easily adapted and used by researchers and industries everywhere.

 描述（由应用程序提供）：本应用的长期目标是提供一个高通量平台，以测试化学暴露对人脑发育的影响。在美国，化学物质在将其纳入消费产品之前并不需要健康或环境安全测试。超过80,000种注册化学品正在商业中，其中大多数都没有有关其长期健康影响的任何信息。考虑其中的每一个都是一个缓慢而昂贵的过程。当前，没有适当的系统来筛选化合物在产品实施之前的健康后果，并且有80,000多种注册化学品在商业中，需要一种高通量机制来筛选这些化合物。人类多能干细胞（HPSC）技术提供了一个可拨动的平台来研究大脑发育的早期阶段。在我们实验室的最新研究中，我们调整了一项方案，将HPSC分化为77天的脑皮质神经元，概括了皮质发育的每个阶段。大脑皮层已被证明对发育过程中的化学暴露特别敏感。我们实验室的初步数据表明，该协议早期的化学暴露会导致在开发过程中不同时间的关键神经发展基因的变化，提供证据表明我们的体外系统可用于评估化学毒性。在这里，我们建议使用HPSC技术开发高通量测定法，以评估它们如何改变人脑发育，以筛选大量化合物。这项研究将开发一种具有成本效益的工具，可以通过简单的PCR分析分析数千种化合物，该化合物很容易被各地的研究人员和行业使用。