Causal mechanisms of anesthetic induction and emergence in human cortical organoids

人类皮质类器官麻醉诱导和苏醒的因果机制

• 批准号: 10752425
• 负责人: Daniel Toker
• 金额: $ 7.18万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752425
• 关键词: 3-Dimensional; Acetylcholine; Aminobutyric Acids; Anesthesia procedures; Anesthetics; Animal Model; Area; Assessment tool; Awareness; Brain; Cerebrum; Chemosensitization; Clinical; Coma; Complex; Conscious; Delirium; Development; Dopamine; Embryo; Entropy; Event; Exhibits; Exposure to; Feedback; Foundations; Frequencies; General Anesthesia; Histamine; Human; Hypothalamic structure; In Vitro; Intervention; Lateral; Length of Stay; Mediating; Modeling; Molecular; Neuroglia; Neurons; Norepinephrine; Organoids; Pathologic; Patients; Persistent Vegetative State; Play; Process; Propofol; Protocols documentation; Recovery; Research; Role; Structure; System; Testing; Thalamic structure; Time; Unconscious State; Vegetative States; awake; basal forebrain; care costs; cholinergic; excitatory neuron; experience; high throughput screening; human pluripotent stem cell; human tissue; hypocretin; improved; in vivo; induced pluripotent stem cell; inhibitory neuron; locus ceruleus structure; mammilloinfundibular nucleus structure; method development; neural; neural circuit; neuroregulation; noradrenergic; novel; novel therapeutics; phenomenological models; raphe nuclei; receptor; respiratory; screening; tool

PROJECT SUMMARY This project aims to use human cortical organoids, which are cortex-like structures generated in vitro from human induced pluripotent stem cells (hiPSCs), to resolve outstanding questions in our understanding of causal mechanisms underlying the mesoscale phenomenology of anesthetic induction (AI) and anesthetic emergence (AE). Millions of patients undergo general anesthesia every year, but the mechanisms by which anesthetic drugs give rise to the hallmarks of AI remain unresolved. Even less well understood are the mechanisms by which the brain emerges from anesthesia - a process over which clinicians have almost no control, and which is frequently associated with complications such as emergence delirium, respiratory events, and delayed emergence, which results in prolonged hospital stays and increased cost of care. In addition, 1-2 per every 1000 patients will experience intraoperative awareness with explicit recall, for reasons that are not understood. While a number of hypotheses regarding the mechanisms of AI and AE have been put forward, these hypotheses are still debated because of complex inter-circuit interactions during AI and AE in the intact brain. In particular, it is widely believed that the major cause of AI is the potentiation of cortical GABAa receptors, but it has been difficult to disentangle the effects of cortical GABAa potentiation from the subcortical effects of anesthesia, which may likewise contribute to AI. Similarly, though it is generally believed that at least one, if not several cortically projecting neuromodulatory structures - including the histaminergic tuberomammillary nucleus of the hypothalamus, the cholinergic basal forebrain, the serotonergic raphe nuclei, the orexinergic lateral hypothalamus, and the noradrenergic locus coeruleus - directly drive emergence from anesthesia, these systems are densely interconnected and mutually excitatory. For this reason, in vivo research has been unable to resolve which, if any, of these systems directly cause AE. A promising but completely unexplored tool for resolving these questions are human cortical organoids. Our team has recently developed a protocol for fusing together networks of excitatory and inhibitory cortical-like neurons derived from hiPSCs. These fusion cortical organoids can recapitulate the oscillatory electric activity of the awake human cortex, and our preliminary results suggest that these cortical organoids can mimic the mesoscale hallmarks of AI when they are exposed to the anesthetic propofol. Importantly, cortical organoids consist of purely cortical- like human tissue, and lack any influence from subcortical structures or neuromodulatory systems. This allows us to use human cortical organoids to isolate cortical versus non-cortical causal mechanisms of both AI and AE. Successful modeling of AI and AE in brain organoids would illustrate the utility of these structures in high- throughput screening of novel drugs for inducing anesthesia or emergence from anesthesia, potentially even on a single-patient basis. Additionally, this project would establish the potential for human brain organoids in screening therapies for other states of unconsciousness, such as coma and persistent vegetative states.

项目摘要 该项目旨在使用人类皮质器官，它们是从体外产生的类似皮质的结构 人类引起的多能干细胞（HIPSC），以解决我们对 麻醉诱导（AI）和麻醉的中尺度现象学的因果机制 出现（AE）。每年有数百万患者每年接受全身麻醉，但其机制 麻醉药引起了AI的标志，但仍未解决。甚至不太了解的是 大脑从麻醉中出现的机制 - 临床医生几乎没有的过程 控制，并且经常与并发症有关，例如出现del妄，呼吸事件， 延迟出现，导致住院时间长时间和增加的护理成本。另外，1-2 每1000名患者都会经历术中意识，并明确召回 理解。尽管已经提出了许多关于AI和AE机制的假设，但 这些假设仍然是争议的 脑。特别是，人们普遍认为AI的主要原因是皮质Gabaa的增强 受体，但是很难将皮质Gabaa增强的影响与皮层下层脱离 麻醉的作用，这可能同样有助于AI。同样，尽管通常认为至少 一种，即使不是几个皮质投射的神经调节结构 - 包括组胺能 下丘脑的结核核核，胆碱能基的前脑，血清素能raphe核， 下丘脑的甲脱甲素能和去甲肾上腺素的基因座 - 直接驱动出现 麻醉，这些系统是密集的互连和相互兴奋的。因此，体内 研究无法解决，这些系统中的这些系统直接引起AE。有希望的，但是 解决这些问题的完全未开发的工具是人类皮质器官。我们的团队最近有 开发了一种协议，将其融合在一起的兴奋性和抑制性皮质样神经元网络。 hipscs。这些融合皮质器官可以概括醒着人的振荡电活动 皮质，我们的初步结果表明，这些皮质类器官可以模仿中尺度的标志 AI暴露于麻醉丙泊酚时。重要的是，皮质器官由纯粹的皮质 - 像人体组织一样，缺乏皮层结构或神经调节系统的任何影响。这允许 美国使用人体皮质器官分离AI和AI的非皮质因果机制 Ae。在脑器官中的AI和AE的成功建模将说明这些结构在高位中的实用性 新药物的吞吐量筛查，以诱导麻醉或出现麻醉，甚至可能 在单一的基础上。此外，该项目将确定人类脑器官的潜力 筛查其他无意识状态，例如昏迷和持续的营养状态。