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类器官，这是体外产生的皮质样结构， 人类诱导多能干细胞（hiPSC），以解决我们理解的悬而未决的问题， 麻醉诱导（AI）和麻醉诱导的中尺度现象学的因果机制 出现（AE）。每年有数百万患者接受全身麻醉，但其机制 麻醉药物引起AI的标志仍然没有解决。更不容易理解的是， 大脑从麻醉中出现的机制-临床医生几乎没有 控制，并经常与并发症，如出现谵妄，呼吸事件， 以及延迟出现，这导致住院时间延长和护理成本增加。此外，1-2 每1000例患者中有1例将经历术中意识并明确回忆，原因不是 明白虽然已经提出了许多关于AI和AE机制的假设， 这些假设仍然存在争议，因为在完整的脑内，AI和AE期间存在复杂的回路间相互作用， 个脑袋特别是，人们普遍认为AI的主要原因是皮层GABA a的增强 受体，但很难将皮层GABA a增强的作用与皮层下GABA受体的作用分开。 麻醉的影响，这也可能有助于AI。同样，尽管人们普遍认为， 一个，如果不是几个皮质投射神经调节结构-包括组胺能 下丘脑的结节乳头核，胆碱能基底前脑，多巴胺能中缝核， 食欲素能的外侧下丘脑和去甲肾上腺素能的蓝斑直接驱动从 麻醉，这些系统紧密相连，相互兴奋。因此，在体内 研究一直无法解决这些系统中的哪些（如果有的话）直接导致AE。一个有前途的，但 解决这些问题的完全未开发的工具是人类皮质类器官。我们的团队最近 开发了一种将兴奋性和抑制性皮质样神经元网络融合在一起的协议， hiPSC。这些融合的皮质类器官可以重现清醒的人类的振荡电活动 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， 当他们接触到麻醉剂异丙酚时。重要的是，皮质类器官由纯粹的皮质- 就像人体组织一样，没有任何皮层下结构或神经调节系统的影响。这允许 我们使用人类皮质类器官来分离AI和AI的皮质与非皮质因果机制， AE. AI和AE在脑类器官中的成功建模将说明这些结构在高密度脑中的实用性。 用于诱导麻醉或从麻醉中苏醒的新药的通量筛选，甚至可能 在一个病人的基础上。此外，该项目将建立人脑类器官的潜力， 筛选治疗其他无意识状态，如昏迷和持续植物人状态。