Multimodal Analysis of Gestational Health and Placental Injury in Opioid-Affected Pregnancies

项目总结 阿片类药物在医学上用于安全的止痛和管理。然而，非法阿片类药物的使用大大减少了 在过去十年中，在美国各地有所增加，在新冠肺炎大流行期间进一步恶化，导致 对人类健康产生深远影响。具体地说，怀孕期间的阿片类药物使用障碍(OUD)会造成 与妊娠相关的孕产妇发病率和死亡率、胎儿生长受限(FGR)和 相关并发症、新生儿阿片类药物戒断综合征和长期神经行为影响。其中一些 尽管使用了更安全的阿片类药物，如丁丙诺啡和美沙酮，但这些风险仍然存在。 穆德(Moud)患者。而目前的研究主要集中在阿片类药物经胎盘向胎儿的转运 以及阿片类药物对婴儿的不良影响，非法和处方类阿片类药物对胎盘的直接影响 发展、分化和功能在很大程度上是未被探索的。胎盘漂浮绒毛调节母体- 胎儿气体交换、营养摄取、废物释放、免疫防御和激素和 细胞外小泡(EVS)。这些绒毛被一层多核的、终末分化的 合体滋养层细胞(STB)，它构成胎盘前线，直接接触阿片类药物。 母血。这一层的下层是单核祖细胞滋养层细胞(CTB)，它 通过分化和融合的过程补充机顶盒的层。重要的是，STB和 CTB层与妊娠相关的并发症有关，包括FGR和死产。我们在这里寻找 研究阿片类药物依赖的胎盘损伤，重点关注胎盘最关键和最独特的一层 滋养层细胞。我们将招募OUD参与者，包括非法阿片类药物和Moud(丁丙诺啡， (美沙酮)，检查她们的怀孕过程和产后第一年孩子的健康状况。 使用来自每个参与者的生物标本，包括三个三个月的孕妇血浆和尿液， 胎盘活检和胎儿脐带血分娩时，我们将采用多模式尖端技术， 包括单细胞RNAseq、空间转录学、蛋白质芯片细胞学和胎盘EV RNA图谱， 并探讨阿片类药物对母体-胎盘-胎儿三联体的分子和细胞过程的影响。 生态系统。为了从机制上深入了解基因表达和EV货物的功能变化，我们将 使用一系列模型系统，包括人类滋养层干细胞和培养的原代人类 滋养细胞，并机械地询问阿片类药物损伤的潜在途径。我们将进一步关联密钥 具有临床评估的分子特征，包括孕妇妊娠障碍、围产期和婴儿 神经发育结果。在我们跨学科团队的支持下，我们的战略计划共同实现了 美国将解决与阿片类药物影响的人类胎盘生物学相关的至关重要的知识空白 怀孕。

PROJECT SUMMARY Opioids are medically used for safe pain relief and management. However, illicit opioid use has substantially increased across the US in the past decade, with further worsening during the COVID-19 pandemic, leading to a profound impact on human health. Specifically, opioid use disorder (OUD) during pregnancy poses an increased risk of pregnancy-associated maternal morbidity and mortality, fetal growth restriction (FGR) and related complications, neonatal opioid withdrawal syndrome, and long-term neurobehavioral effects. Some of these risks persist despite the use of safer opioids, such as buprenorphine and methadone, medications for OUD (MOUD) patients. Whereas current studies center mainly on transplacental opioid transport to the fetus and the adverse effects of opioids on infants, the direct impact of illicit and prescription opioids on placental development, differentiation, and function are largely unexplored. The placental floating villi mediate maternal- fetal gas exchange, nutrient uptake, waste release immune defense and the production of hormones and extracellular vesicles (EVs). These villi are covered by a layer of multinucleated, terminally differentiated syncytiotrophoblasts (STBs), which forms the feto-placental frontline that is directly exposed to opioids in the maternal blood. Subjacent to this layer are mononucleated, progenitor cytotrophoblasts (CTBs), which replenish the STB layer through the process of differentiation and fusion. Importantly, injuries to the STB and CTB layers are implicated in pregnancy-associated complications, including FGR and stillbirth. Here we seek to investigate opioid-dependent placental injury, focusing on the most critical and unique layer of placental trophoblasts. We will enroll participants with OUD, including illicit opioids and MOUD (buprenorphine, methadone), examine their pregnancy course and their children’s health through the first year postpartum. Using biospecimens from each participant, including maternal plasma and urine across the three trimesters, placental biopsies and fetal cord blood at delivery, we will employ multimodal cutting-edge technologies, including single-cell RNAseq, spatial transcriptomics, protein chip cytometry and placenta EV RNA profiling, and explore the molecular and cellular processes affected by opioids in the maternal-placental-fetal trio- ecosystem. To gain mechanistic insights into the functional changes in gene expression and EV cargo, we will use an array of model systems, including human trophoblast stem cells and cultured primary human trophoblasts, and mechanistically interrogate pathways underlying opioid injury. We will further correlate key molecular signatures with clinical assessment, including maternal gestational disorders, perinatal and infant neurodevelopmental outcomes. Together, our strategic plan, bolstered by our transdisciplinary team, enables us to address critically important knowledge gaps related to human placenta biology in opioid-affected pregnancies.