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.

摘要 在过去的十年中，阿片类药物使用障碍在孕妇和10岁以下的妇女中显著增加。 生育年龄与此同时，我们看到新生儿阿片类药物戒断综合征（NOWS）急剧上升。 尽管NOWS具有明确的短期表型症状，但其分子效应和长期效应可能与NOWS相关。 后果是难以理解的。这一点尤其令人关切，因为仅阿片类药物暴露，以及 戒断，可以诱导大脑炎症，最近的工作表明，促炎性增加， 细胞因子和趋化因子，这可能是促进反应性神经胶质细胞作为一个结果，在子宫内阿片类药物暴露。 此外，在子宫内的临床前模型中已经观察到认知功能和记忆缺陷。 在临床中观察到阿片类药物暴露、智力障碍和注意力障碍增加。 以前被诊断为NOWS的人群。虽然大部分研究都集中在炎症和炎症反应上， 子宫内阿片类药物暴露在与奖励和认知相关的大脑区域，对大脑的了解较少 NOWS中与戒断症状相关的区域，以及这可能与后来的认知功能有关。 生活了解子宫内阿片类药物暴露和戒断的促炎作用可能会导致新的 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，该模型先前已显示炎症标志物的上调， 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， 蓝斑，并确定对戒断行为指数和随后的认知功能的影响。更 具体而言，在NOWS大鼠模型中，我们将利用免疫组织化学、qPCR和多重测定， 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，以及前 炎性细胞因子和相关神经肽，以及它们各自与定量 行为戒断症状此外，我们将测试反应性胶质细胞在物理上的具体作用。 通过全身给予神经胶质抑制剂米诺环素在新生大鼠中的戒断症状。 我们将在PN21和PN60评估由此产生的认知功能，此时大鼠被视为青春期前大鼠。 和成年早期。我们将测试海洛因或盐水暴露大鼠的空间学习和记忆 在PN 10时出现突然戒断的幼仔。其他实验还将包括使用 新生儿全身给予米诺环素以减轻先前在临床前研究中观察到的认知缺陷 和NOWS的临床人群。总之，这些实验将使我们能够更好地了解 炎症对躯体戒断症状的影响，以及与躯体戒断的关系 症状和随后的认知功能，由于在子宫内阿片类药物暴露和沉淀 从而为NOWS的治疗建立新的目标，并为未来的治疗奠定基础。 评估其长期影响的研究。