Evaluation of inflammation in the locus coeruleus during physical withdrawal symptoms and cognitive development in a rat model of neonatal opioid withdrawal syndrome (NOWS)

新生儿阿片戒断综合征 (NOWS) 大鼠模型身体戒断症状和认知发展过程中蓝斑炎症的评估

• 批准号: 10750776
• 负责人: Sara Lynn Mills-Huffnagle
• 金额: $ 3.7万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750776
• 关键词: Address; Adult; Amygdaloid structure; Animal Model; Attention; Attenuated; Behavioral; Birth; Brain; Brain Stem; Brain region; Characteristics; Child; Clinical; Cognition; Cognitive; Cognitive deficits; Compensation; Corpus striatum structure; Cost of Illness; Diagnosis; Disease; Evaluation; Exposure to; Female of child bearing age; Foundations; Future; Genetic Induction; Heroin; Heroin Dependence; Hippocampus; Hyperactivity; Immunohistochemistry; Infant; Inflammation; Inflammatory; Intellectual impairment; Intervention; Learning; Life; Long-Term Effects; Longitudinal Studies; Measures; Medial; Medical Care Costs; Memory; Memory impairment; Microglia; Minocycline; Modeling; Molecular; Molecular Target; Motor Activity; Mutation; Naloxone; Neonatal; Neonatal Abstinence Syndrome; Neuroglia; Neurons; Neuropeptides; Neurotransmitters; Norepinephrine; Operative Surgical Procedures; Opioid; Pennsylvania; Phenotype; Population; Pre-Clinical Model; Pregnancy; Pregnant Women; Proteins; Publishing; Rattus; Regulation; Research; Rewards; Rodent Model; Role; Saline; Source; Symptoms; Syndrome; System; TLR4 gene; Techniques; Testing; Training; Translational Research; Ultrasonics; United States; Up-Regulation; Weaning; Withdrawal; Withdrawal Symptom; Work; career; chemokine; clinically relevant; cognitive development; cognitive function; cognitive testing; cohort; cost; cytokine; education cost; emerging adult; endogenous opioids; experience; experimental study; in utero; indexing; inflammatory marker; inhibitor; insight; interest; locus ceruleus structure; mu opioid receptors; multiplex assay; neonatal brain development; neonate; novel; opioid exposure; opioid use; opioid use disorder; opioid withdrawal; physical symptom; postnatal; pre-clinical; preadolescence; pup; stem; targeted treatment; treatment strategy; vocalization

Abstract In the past decade, opioid use disorders have significantly increased in pregnant women and women of childbearing age. In parallel, we have seen a drastic rise in neonatal opioid withdrawal syndrome (NOWS). Despite having well-defined, short-term phenotypic symptoms for NOWS, molecular effects and long-term consequences are incomprehensible. This is of particular concern given that opioid exposure alone, as well as withdrawal, can induce inflammation in the brain, and recent work has shown an increase in pro-inflammatory cytokines and chemokines, which may be promoted by reactive glia as a result of in utero opioid exposure. Moreover, cognitive functioning and memory deficits have been observed in preclinical models of in utero opioid exposure, and intellectual impairments and increased attention disorders have been observed in clinical populations previously diagnosed with NOWS. While much of the research has focused on inflammation and in utero opioid exposure in brain regions associated with reward and cognition, less is known about the brain regions associated with withdrawal symptoms in NOWS, and how this may relate to cognitive function later in life. Understanding the pro-inflammatory effects of in utero opioid exposure and withdrawal may allow for novel targets for the treatment of NOWS, and better understanding of long-term consequences. Therefore, by using an established rat model of NOWS that has previously shown an upregulation of inflammatory markers due to in utero opioid exposure, we will investigate inflammation in a withdrawal-associated brain region, the locus coeruleus, and determine effects on behavioral indices of withdrawal and subsequent cognitive function. More specifically, in a rat model of NOWS, we will utilize immunohistochemistry, qPCR, and multiplex assays to evaluate inflammatory markers including reactive glial cells, toll-like receptor 4 expression, and levels of pro- inflammatory cytokines and associated neuropeptides, and their respective associations with quantified behavioral withdrawal symptoms. In addition, we will test the specific role of reactive glia on physical withdrawal symptoms in the rat neonate by systemically administering the glia-inhibiting agent, minocycline. We will assess the resultant cognitive function at PN21 and PN60, when rats are considered preadolescent and early adulthood, respectively. We will test spatial learning and memory in heroin- or saline-exposed rat pups who experienced precipitated withdrawal at PN10. Additional experiments will also include the use of neonatal, systemic minocycline administration to mitigate cognitive deficits previously observed in preclinical and clinical populations of NOWS. Together, these experiments will allow us to better understand the effects of inflammation on physical withdrawal symptoms, as well as the relationship between physical withdrawal symptoms and subsequent cognitive function, as a result of in utero opioid exposure and precipitated withdrawal, thereby establishing novel targets for the treatment of NOWS and laying the foundation for future studies evaluating its long-term consequences.

抽象的 在过去的十年中，孕妇和妇女中阿片类药物使用障碍显着增加 生育年龄。与此同时，我们发现新生儿阿片类药物戒断综合征 (NOWS) 急剧上升。 尽管NOWS 有明确的短期表型症状，但分子效应和长期 后果是难以理解的。鉴于仅接触阿片类药物以及阿片类药物，这一点尤其值得关注 戒断，会引起大脑炎症，最近的研究表明促炎细胞增多 细胞因子和趋化因子，由于子宫内阿片类药物暴露，反应性神经胶质细胞可能会促进这些因子和趋化因子的产生。 此外，在子宫内的临床前模型中观察到认知功能和记忆缺陷 临床中已观察到阿片类药物暴露、智力障碍和注意力障碍增加 之前诊断出患有NOWS 的人群。虽然大部分研究都集中在炎症和 与奖赏和认知相关的大脑区域的子宫内阿片类药物暴露，对大脑知之甚少 NOWS 中与戒断症状相关的区域，以及这与后来的认知功能有何关系 生活。了解子宫内阿片类药物暴露和戒断的促炎作用可能有助于新的研究 治疗NOWS 的目标，以及更好地了解长期后果。因此，通过使用 一种已建立的NOWS 大鼠模型，该模型先前已显示出由于以下因素导致的炎症标记物的上调 在子宫内阿片类药物暴露中，我们将研究与戒断相关的大脑区域的炎症，该区域 蓝藻，并确定对戒断行为指数和随后认知功能的影响。更多的 具体来说，在 NOWS 大鼠模型中，我们将利用免疫组织化学、qPCR 和多重检测来 评估炎症标志物，包括反应性神经胶质细胞、Toll 样受体 4 表达以及促炎症因子水平 炎症细胞因子和相关神经肽，以及它们各自与定量的关联 行为戒断症状。此外，我们还将测试反应性胶质细胞对身体的具体作用。 通过系统地给予神经胶质细胞抑制剂米诺环素，使新生大鼠出现戒断症状。 当大鼠被视为青春期前时，我们将评估 PN21 和 PN60 时产生的认知功能 和成年早期，分别。我们将测试暴露于海洛因或盐水的大鼠的空间学习和记忆 在 PN10 时经历过突然退缩的幼犬。额外的实验还将包括使用 新生儿全身使用米诺环素以减轻先前在临床前观察到的认知缺陷 和NOWS 的临床人群。总之，这些实验将使我们能够更好地了解 炎症对身体戒断症状的影响，以及身体戒断之间的关系 由于子宫内阿片类药物暴露和沉淀而导致的症状和随后的认知功能 退出，从而建立NOWS治疗的新靶点，为未来奠定基础 研究评估其长期后果。