lncRNAs and Anesthetic-Induced Developmental Neurotoxicity

项目总结 2016年，美国食品和药物管理局警告说，反复或长期使用全身麻醉药 三岁以下的儿童可能会影响他们的大脑发育。这一警告引起了严重的关注。 关于儿科麻醉的安全性。麻醉剂研究领域主要有两个障碍-- 诱发发育性神经毒性(AIDN)：1)到目前为止，AIDN的证据大多来自 动物研究。人体研究的结果仍然没有定论。2)其机制在很大程度上是未知的。 这项研究的目标是通过使用三维(3D)的体外系统来解决这两个障碍 诱导多能干细胞和小鼠模型生成的人迷你大脑研究新事物 长非编码RNA(LncRNAs)的机制。人类迷你脑更接近于发育中的人类 大脑，无论在结构上还是功能上，都超过了广泛使用的2D神经元。我们的初步数据提供了 首次有证据表明，临床上相关剂量的异丙酚和七氟醚，两种儿科常用药物 麻醉剂，在人类迷你大脑中诱导急性细胞死亡。我们还发现新生儿接触异丙酚 改变小鼠的lncRNA表达谱并引起多种不良反应(E/I失衡，神经元死亡， 和记忆功能受损)。我们令人兴奋的发现表明，异常表达的lncRNA可能 为AIDN做出贡献。因此，我们将探讨lncRNAs在AIDN中的作用和机制 使用人类迷你脑和小鼠模型的神经元活动和网络。HyperCam Alpha 通过提供高吞吐量、详细的数据，设备将显著提高我们的生产率和产能 收集我们对3D人类迷你脑和小鼠组织切片的电生理学研究。这项建议 有望提供新的分子和电生理机制方面的见解 儿科麻醉剂暴露的神经发育后果。这将进一步有助于发展 更合理的神经保护策略与儿科麻醉药的使用有关，并朝着更好的方向发展 儿科麻醉使用的安全性保证。

PROJECT SUMMARY In 2016, the U.S. Food and Drug Administration warned that repeated or lengthy use of general anesthetics in children below the age of three might affect their brain development. This warning raises serious concerns regarding the safety of pediatric anesthesia. There are two main barriers in the research field of anesthetic- induced developmental neurotoxicity (AIDN): 1) So far, most of the evidence for AIDN was obtained from animal studies. The results from human studies remain inconclusive. 2) The mechanisms are largely unknown. The goal of this study is to address both of these barriers by using an in vitro system of three-dimensional (3D) human mini brains generated from induced pluripotent stem cells and mouse models to study novel mechanisms of long non-coding RNAs (lncRNAs). Human mini brains are more similar to developing human brains, both structurally and functionally, than the widely used 2D neurons. Our preliminary data provided the first evidence showing that clinically relevant doses of propofol and sevoflurane, two commonly used pediatric anesthetics, induced acute cell death in human mini brains. We also found that neonatal propofol exposure altered lncRNA expression profiles and caused multiple adverse effects in mice (E/I imbalance, neuronal death, and impaired memory function). Our exciting findings suggest that the abnormally expressed lncRNAs might contribute to AIDN. Thus, we will examine the role and mechanism of lncRNAs in AIDN such as abnormal neuronal activity and networks using both human mini brain and mouse models. The HyperCAM Alpha equipment would significantly improve our productivity and capacity by enabling high throughput, detailed data collection of our electrophysiological studies on 3D human mini brains and mouse tissue slices. This proposal is expected to provide new molecular and electrophysiological mechanistic insights into the neurodevelopmental consequences of pediatric anesthetic exposure. This will further aid in the development of more rational neuroprotective strategies related to pediatric anesthetic use, and movement towards a better assurance of safety for pediatric anesthesia use.