Simultaneous prenatal alcohol and cannabinoid exposure & offspring corticostriatal neurocircuitry

产前同时接触酒精和大麻素

• 批准号: 10615012
• 负责人: Siara Rouzer
• 金额: $ 6.91万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10615012
• 关键词: 2-arachidonylglycerol; ATAC-seq; Actins; Acute; Address; Age; Alcohol consumption; Alcohols; Applications Grants; Behavior; Behavioral; Behavioral Assay; Biological; Biological Assay; CNR1 gene; Cannabinoids; Cannabis; Child; Chromatin; Clinical assessments; Code; Consumption; Corpus striatum structure; Cytoskeletal Proteins; Data; Development; Drug Combinations; Drug Exposure; Drug usage; Economics; Endocannabinoids; Environment; Enzyme-Linked Immunosorbent Assay; Ethanol; Exhibits; Exposure to; Female; Fetal Alcohol Exposure; Fetal Development; Future; Genes; Genetic; Glutamates; Goals; Growth; Health; Human; Hyperactivity; Impairment; Individual; Inhalation; Knowledge; Ligands; Link; Literature; Long-Term Depression; Longevity; Mentors; Mentorship; Messenger RNA; Modification; Mothers; Motor; Mus; Neurologic; Neurons; Nuclear; Output; Patient Self-Report; Pharmaceutical Preparations; Physiological; Pre-Clinical Model; Pregnancy; Proteins; Proteomics; Psychotropic Drugs; Quality of life; Recording of previous events; Records; Research; Research Personnel; Risk; Self Administration; Signal Pathway; Societies; Source; Synapses; Synaptic Transmission; Synaptic plasticity; Synaptosomes; Technical Expertise; Teratology; Testing; Therapeutic; Therapeutic Intervention; Training; Transposase; adverse outcome; alcohol exposure; alcohol seeking behavior; anandamide; behavioral impairment; behavioral phenotyping; brain tissue; career; child bearing; design; developmental disease; disability; drug seeking behavior; endocannabinoid signaling; endogenous cannabinoid system; experimental study; fetal; fetal drug exposure; fetal marijuana exposure; field study; gene network; improved; in utero; locomotor deficit; male; marijuana user; motor deficit; motor disorder; motor impairment; nerve stem cell; neural circuit; neurodevelopment; neurogenesis; neurophysiology; offspring; pre-clinical; preference; prenatal; prenatal exposure; presynaptic; protein expression; psychosocial; receptor; sex; social stigma; synthetic cannabinoid; targeted treatment; transcriptome sequencing; young adult

PROJECT SUMMARY Prenatal alcohol exposure (PAE) is the most common cause of developmental disorders, though concurrent exposure to other psychoactive substances may exacerbate adverse outcomes. Recent preference for simultaneous use of alcohol and cannabis (SAC) among young adults of child-bearing age combined with early preclinical evidence for synergistic developmental harm, support a premise for investigating SAC as a specific developmental source of disability. Despite indications that both PAE and prenatal cannabinoid exposure (PCE) alter endocannabinoid signaling through cannabinoid receptor 1 (CNR1) in offspring, it is yet unknown whether impairments to CNR1 – a receptor essential to healthy fetal neurodevelopment – underly impairments associated with SAC. Notably, PAE impairs CNR1-regulated synaptic transmission from striatal-projecting cortical neurons. In drug-naïve mice, impaired striatal CNR1-regulated activity contributes to motor dysfunction, hyperactivity and increased drug-seeking behaviors, deficits observed in humans with PAE and PCE. Therefore, the objective of this proposal is to investigate the impact of SAC on CNR1-associated neural circuits and behaviors in exposed offspring. Preliminary data from our lab have shown that a CNR1-associated gene network that regulates striatal synaptic activity is changed by acute fetal ethanol exposure, and combined SAC exposure augments growth deficits in neural stem cells from single-drug and drug-free exposures. Collectively, the literature and these data inform our central hypothesis that SAC offspring will exhibit impaired CNR1-linked synaptic mechanisms within the striatum corresponding with increased motor deficits and drug-seeking behaviors. We will test this hypothesis in the following aims: Specific Aim 1) To assess SAC-induced changes in genes that regulate corticostriatal synaptic activity. We will use integrative RNAseq and ATAC-seq to investigate a pre- determined gene network associated with CNR1-regulated corticostriatal synaptic plasticity in prenatally exposed offspring. Specific Aim 2) To assess whether SAC augments behavioral deficits and striatal protein expression. We will perform a battery of behavioral assays investigating native deficits in motor function and ethanol-seeking behaviors in prenatally exposed offspring. We will compare behavioral deficits with striatal protein expression using sandwich ELISAs, quantifying CNR1 and endogenous cannabinoid ligands anandamide and 2-arachidonylglycerol. Prenatal drug exposure will occur from gestational days 12-15, a period of peak neurogenesis for corticostriatal neurons, and will incorporate vaporized ethanol inhalation and i.p administration of synthetic cannabinoid CP-55940. Successful completion of this proposal will identify specific mechanistic changes underlying SAC, a translationally-relevant but under-investigated form of prenatal drug exposure, and inform future targets for therapeutic intervention. Furthermore, the proposed experiments will provide the framework for an excellent training environment, including mentorship from multiple sponsors and collaborators, and will subsequently inform a scientifically rigorous K99/R00 grant application.

项目概要 产前酒精暴露（PAE）是发育障碍的最常见原因，尽管同时存在 接触其他精神活性物质可能会加剧不良后果。最近偏好 育龄年轻人同时使用酒精和大麻（SAC） 协同发育危害的临床前证据支持将 SAC 作为特定的研究的前提 残疾的发育根源。尽管有迹象表明 PAE 和产前大麻素暴露 (PCE) 通过大麻素受体 1 (CNR1) 改变后代的内源性大麻素信号传导，目前尚不清楚是否 CNR1 的损伤（一种对健康胎儿神经发育至关重要的受体） - 相关的潜在损伤 与 SAC。值得注意的是，PAE 会损害纹状体投射皮质神经元的 CNR1 调节的突触传递。 在未接受药物治疗的小鼠中，纹状体 CNR1 调节的活性受损会导致运动功能障碍、多动症和 PAE 和 PCE 患者的寻药行为增加以及缺陷。因此，目标是 该提案旨在研究 SAC 对暴露于环境中的 CNR1 相关神经回路和行为的影响 后代。我们实验室的初步数据表明，调节纹状体的 CNR1 相关基因网络 急性胎儿乙醇暴露会改变突触活动，并且联合 SAC 暴露会促进生长 单一药物和无药物暴露导致的神经干细胞缺陷。总的来说，文献和这些数据 告知我们的中心假设，即 SAC 后代将表现出受损的 CNR1 相关突触机制 纹状体与运动缺陷和寻求药物行为的增加相对应。 我们将在以下目标中检验这一假设： 具体目标 1) 评估 SAC 诱导的基因变化 调节皮质纹状体突触活动。我们将使用整合 RNAseq 和 ATAC-seq 来研究预 确定与产前 CNR1 调节的皮质纹状体突触可塑性相关的基因网络 暴露的后代。具体目标 2) 评估 SAC 是否会增强行为缺陷和纹状体蛋白 表达。我们将进行一系列行为分析，调查运动功能的天然缺陷和 产前暴露的后代的乙醇寻求行为。我们将行为缺陷与纹状体进行比较 使用夹心 ELISA 进行蛋白质表达，定量 CNR1 和内源性大麻素配体 花生四烯酸乙醇胺和2-花生四烯酸甘油。产前药物暴露发生在妊娠第 12-15 天，即妊娠期 皮质纹状体神经元的神经发生峰值，并将结合汽化乙醇吸入和腹膜内注射 合成大麻素 CP-55940 的给药。成功完成该提案将确定具体的 SAC 的机制变化，一种与翻译相关但研究不足的产前药物形式 暴露，并为未来的治疗干预目标提供信息。此外，拟议的实验将 提供良好培训环境的框架，包括来自多个赞助商的指导和 合作者，随后将通知科学严谨的 K99/R00 资助申请。

项目成果

The importance of promoting scientific advocacy & outreach for trainees.

• DOI: 10.1038/s41386-023-01530-6
• 发表时间: 2023-04
• 期刊: NEUROPSYCHOPHARMACOLOGY
• 影响因子: 7.6
• 作者: Rouzer, Siara Kate;Kalinowski, Leanna Marie;Kaseda, Erin Taniyo
• 通讯作者: Kaseda, Erin Taniyo

Microbiota and nutrition as risk and resiliency factors following prenatal alcohol exposure.

• DOI: 10.3389/fnins.2023.1182635
• 发表时间: 2023
• 期刊: FRONTIERS IN NEUROSCIENCE
• 影响因子: 4.3
• 作者: Upreti, Deepa;Rouzer, Siara K. K.;Bowring, Abigail;Labbe, Emma;Kumar, Rosaline;Miranda, Rajesh C. C.;Mahnke, Amanda H. H.
• 通讯作者: Mahnke, Amanda H. H.