Effect of methadone on the developmental properties of human brain organoids

美沙酮对人脑类器官发育特性的影响

• 批准号: 10618375
• 负责人: Gabriel G Haddad
• 金额: $ 64.74万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-15 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10618375
• 关键词: 3-Dimensional; Address; Affect; Astrocytes; Blood; Brain; Buprenorphine; Cessation of life; Cognitive; Data; Dendritic Spines; Dependence; Development; Disease; Drug abuse; Electrophysiology (science); Emergency department visit; Epidemic; Ethics; Fentanyl; Fetal Development; Fetus; Genetic; Glucose; Glycoproteins; Heroin; Hormones; Human; Imaging Techniques; Impairment; Investigation; Life; Maternal Exposure; Membrane; Methadone; Methods; Molecular; Morbidity - disease rate; Mothers; Neonatal Abstinence Syndrome; Neurites; Neurons; Opiate Addiction; Opioid; Organoids; Patch-Clamp Techniques; Pattern; Pregnancy; Pregnant Women; Property; Regulation; Societies; Structure; Synapses; Synaptic Transmission; Synaptic plasticity; Techniques; Thrombospondin 1; Youth; excitatory neuron; fentanyl abuse; fetal; heroin use; human fetal brain; imaging study; in utero; inhibitory neuron; maternal opioid use; mortality; multi-electrode arrays; multiple drug use; neural network; new technology; new therapeutic target; novel; offspring; opioid abuse; opioid epidemic; opioid exposure; opioid overdose; opioid use in pregnancy; overdose death; patch clamp; postsynaptic; prescription opioid; presynaptic; productivity loss; receptor; statistics; synaptic function; synaptogenesis

ABSTRACT The opioid crisis has become a national epidemic and the statistics are startling. In the past decade there has been a sharp increase in heroin use, opiate prescriptions and fentanyl abuse. Overdose deaths have doubled nationally since 2000 and in 2015, and more than 33,000 deaths were attributable to overdose from opioids. In addition, there has been a major increase between 1998 and 2011 in the number of opioid-dependent pregnant women, such as with methadone dependency. Although opioids have been well studied in general, the epidemic of opioid abuse, especially in pregnant women, has unmasked how little we know about the effect of methadone on fetal brain development. In the past several years, we have taken advantage of a newer technology, the 3D- brain organoids, that facilitated enormously the investigation of early human brain development. This has provided us with an unprecedented opportunity to investigate the cellular and molecular mechanisms underlying the effect of opioids on early brain development. Using such methods, we have been able to produce exciting preliminary data showing that methadone decreases synaptic transmission and possibly affects synaptic plasticity. Based on our recent results, we have posed the following overall hypothesis: Opioid exposure leads to abnormal synaptogenesis and impaired synaptic transmission during fetal brain development. In order to address this hypothesis, we have formulated the following Specific Aims: Specific Aim 1: To determine the effect of methadone on neural network activity during development in human cortical organoids. We will use multi- electrode array recordings to explore how methadone modifies the neural network activity. Specific Aim 2: To determine the effect of methadone on cellular electrophysiological properties and synaptic function and structure during development in human cortical organoids. We will investigate AP firing properties, synaptic currents, and Na+ and K+ currents in neurons and dissect the pre- and postsynaptic mechanisms using patch-clamp, molecular and imaging techniques. Specific Aim 3: To dissect the mechanisms of the methadone-induced changes in synaptogenesis and synaptic transmission in human cortical organoids. As thrombospondins 1,2 (TSP1,2), astrocyte-secreted glycoproteins, play a role in neurite outgrowth, dendritic spine and synapse formation, we will study the effect of methadone on TSPs to obtain an understanding of the molecular pathobiology of methadone’s effect on the human fetal brain. Our studies in this application are novel and unique and address the important problem of human brain maldevelopment under the influence of methadone in pregnant women. With an understanding of the mechanisms involved in methadone effect, we believe that we can develop novel therapeutic targets to mitigate the effect of methadone on brain development in early life.

抽象的 阿片类药物危机已成为全国范围内的流行病，统计数据是在过去十年中开始的。 海洛因使用量急剧增加，阿片类药物处方和芬太尼滥用死亡人数增加了一倍。 自 2000 年和 2015 年以来，全国有超过 33,000 人死于阿片类药物过量。 此外，1998年至2011年间，依赖阿片类药物的孕妇人数大幅增加 妇女，例如对美沙酮依赖的妇女 阿片类药物滥用，尤其是孕妇滥用，揭示了我们对美沙酮的影响知之甚少 在过去的几年里，我们利用了一项更新的技术，即 3D- 大脑类器官，极大地促进了早期人类大脑发育的研究。 我们获得了前所未有的机会来研究该效应背后的细胞和分子机制 使用这些方法，我们已经能够产生令人兴奋的初步结果。 数据显示美沙酮会降低突触传递并可能影响突触可塑性。 根据我们最近的结果，我们提出了以下总体假设：阿片类药物暴露会导致 胎儿大脑发育过程中突触发生异常和突触传递受损。 针对这一假设，我们制定了以下具体目标： 具体目标 1：确定效果 我们将使用多种方法来研究美沙酮对人类皮质类器官发育过程中神经网络活动的影响。 电极阵列记录，探索美沙酮如何改变神经网络活动。 确定美沙酮对细胞电生理特性以及突触功能和结构的影响 在人类皮质类器官的发育过程中，我们将研究 AP 放电特性、突触电流和 神经元中的 Na+ 和 K+ 电流，并使用膜片钳、分子技术剖析突触前和突触后机制 具体目标 3：剖析美沙酮引起的变化的机制。 人类皮质类器官中的突触发生和突触传递。作为血小板反应蛋白 1,2 (TSP1,2)， 星形胶质细胞分泌的糖蛋白，在神经突生长、树突棘和突触形成中发挥作用，我们将 研究美沙酮对 TSP 的影响，以了解美沙酮的分子病理学 我们在该应用中的研究是新颖且独特的，并且解决了重要的问题。 孕妇美沙酮影响下的人脑发育不良问题。 了解美沙酮作用的机制后，我们相信我们可以开发治疗新药 目标是减轻美沙酮对生命早期大脑发育的影响。