Administrative Supplement: Opioids Perinatal Repository: Dysregulation of Maternal Immunity During Pregnancy by Pregravid Obesity

Opioid use during pregnancy has risen at an alarming rate in the United States from 3.5 to 8.2 per 1000 deliveries between 2010 and 2017. Maternal opioid use disorder (OUD) results in increased incidence of adverse outcomes, notably preterm birth, miscarriage, delivery complications, and neonatal opioid withdrawal syndrome (NOWS). The mechanisms underlying these adverse outcomes are poorly understood due to a paucity of longitudinal clinical studies. One potential mechanism is that the accumulation of opioids in placental tissues alter the structure and function of the placenta, which consequently alter fetal development. Another potential mechanism is via OUD-mediated inflammatory changes at the maternal-fetal interface and in circulation. Indeed, a healthy pregnancy is dependent on tightly regulated gestational age-dependent changes in immune cell frequencies and activation status that are critical for placentation, fetal growth, fetal tolerance, and successful labor. Disturbances of this “immune clock of pregnancy” can result in adverse outcomes such as preeclampsia and preterm birth. Indeed, chronic opioid use in non-gravid individuals leads to altered systemic inflammation and increased susceptibility to infections. In this application, we will test the central hypothesis that maternal opioid use (OUD) leads to increased inflammation and cellular activation in maternal circulation and in the placenta leading to increased incidence of adverse outcomes. We are exquisitely well positioned to respond to this Notice of Special Interest (HEAL Initiative – Biospecimen Collection in Pregnancy (NOT-DA-23- 005)) because of our [1] access to the Perinatal Assistance and Treatment Home (PATHways) program at the University of Kentucky, a unique clinic dedicated to caring for pregnant women with substance use disorder in Appalachia, the epicenter of the U.S. opioid crisis; [2] innovative and multi-modal experimental approach; [3] longitudinal study design; and [4] multidisciplinary team with expertise in maternal fetal medicine, immunology, and genomics. We propose to collect blood samples longitudinally throughout pregnancy from women with a history of chronic OUD. We will also collect paired umbilical cord blood (UCB) and placental samples at delivery as well as blood samples from neonates with NOWS during their stay in the neonatal abstinence care unit (NACU) to complete the following specific aims: Aim 4: Build a research infrastructure for perinatal opioid use research. We are exceptionally well positioned to fulfill this aim by leveraging the framework of the Perinatal Assistance and Treatment Home (PATHways) program, a unique resource at the University of Kentucky serving pregnant women with substance use disorder as well as the continuum of care covering their neonates suffering from NOWS and hospitalized in the NACU. In this aim, we will isolate and cryopreserve maternal peripheral blood mononuclear cells (PBMC) and plasma from maternal blood, umbilical cord blood, and neonatal blood samples as well as preserve samples for genomic assays. We will also isolate and cryopreserve immune cells harvested from decidua and chorionic villous. Finally, biopsies of placental tissues will be paraffin-embedded and frozen for genomics and histological analyses. Available clinical metadata for each biospecimen will also be curated to facilitate data interpretation. Aim 5: Define the impact of pregravid opioid use on systemic maternal inflammation. Maternal OUD increases several obstetric complications and incidence of infection, thus suggesting heightened systemic inflammation. Indeed, OUD in non-gravid individuals disrupts monocyte and T cell activation, thereby altering production of immune mediators and increasing susceptibility to infection. This aim will test the hypothesis that maternal OUD disrupts the immune clock of pregnancy using a combination of flow cytometry-based functional assays and single cell genomics (RNA and ATAC-sequencing). Aim 6: Uncover the impact of maternal OUD on the placenta. Maternal OUD increases vertical transmission of untreated HIV, delays placenta villous maturation, and increases incidence of preeclampsia, preterm birth, and other obstetric complications, thus suggesting alterations of placental structure and immune landscape. This aim will test the hypothesis that maternal OUD dysregulates placental maturation and exacerbates placental inflammation by performing histological studies, spatial transcriptomics analyses and functional assays. Aim 7: Determine the effect of maternal OUD on fetal/neonatal inflammation. The developmental origin of heath and disease hypothesis suggests that in utero environmental cues lead to reprogramming of the developing fetal immune system with long term consequences for the offspring. Indeed, in utero exposure to opioids alters levels of circulating mediators, PBMC responses to LPS in rodent pups, and cytokine production by human UCB CD4 T cells. Thus, this aim will test the hypothesis that maternal OUD leads to aberrant activation of the fetal immune system using similar approaches as described in Aim 5. Our team has the required expertise to complete the aims described above and address the goals of this NOSI: to “add and/or expand biospecimen collection during and immediately after pregnancy to build prospective pathophysiological models of maternal, neonatal, and infant risk and disease related to prenatal opioid use or exposure during pregnancy”. Findings will be integrated to generate a comprehensive model of the impact of maternal OUD on maternal, placental, and fetal/neonatal inflammatory outcomes.