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

 描述(申请人提供)：这项申请的长期目标是提供一个高通量平台，以测试化学品暴露对人脑发育的影响。在美国，在将化学品纳入消费品之前，不需要进行健康或环境安全测试。超过8万种注册的化学品在商业中，其中大多数在使用时没有任何关于其长期健康影响的信息。逐一考虑这其中的每一个都是一个缓慢而昂贵的过程。目前还没有在产品实施之前根据化合物的健康后果对其进行筛选的系统，而且商业上有80,000多种注册的化学品，因此需要一个高通量的机制来筛选这些化合物。人类多能干细胞(HPSC)技术为研究大脑发育的早期阶段提供了一个可操作的平台。在我们实验室最近的一项研究中，我们采用了一种方案，在77天内将hPSCs分化为大脑皮层神经元，概括了大脑皮层发育的每个阶段。大脑皮层已被证明在发育过程中对化学物质暴露特别敏感。我们实验室的初步数据显示，在该方案的早期阶段，化学暴露会导致关键神经发育基因在发育过程中的不同时间发生变化，这为我们的体外系统可以用于评估化学毒性提供了证据。在这里，我们建议开发一种使用hPSC技术的高通量分析方法，通过评估化合物如何改变人脑发育来筛选大量化合物。这项研究将开发一种具有成本效益的工具，通过一种简单的PCR分析方法来分析数千种化合物，这种方法可以很容易地被各地的研究人员和行业改装和使用。

 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.