Placental cytochrome P450 methylomes in infants exposed to prenatal opioids: exploring the effects of neonatal opioid withdrawal syndrome on health horizons

Abstract

Background: Neonatal opioid withdrawal syndrome (NOWS), arises due to increased opioid use during pregnancy. Cytochrome P450 (CYP) enzymes play a pivotal role in metabolizing a wide range of substances in the human body, including opioids, other drugs, toxins, and endogenous compounds. The association between CYP gene methylation and opioid effects is unexplored and it could offer promising insights. Objective: To investigate the impact of prenatal opioid exposure on disrupted CYPs in infants and their anticipated long-term clinical implications. Study Design: DNA methylation levels of CYP genes were analyzed in a cohort of 96 placental tissues using Illumina Infinium MethylationEPIC (850 k) BeadChips. This involved three groups of placental tissues: 32 from mothers with infants exposed to opioids prenatally requiring pharmacologic treatment for NOWS, 32 from mothers with prenatally opioid-exposed infants not needing NOWS treatment, and 32 from unexposed control mothers. Results: The study identified 20 significantly differentially methylated CpG sites associated with 17 distinct CYP genes, with 14 CpGs showing reduced methylation across 14 genes (CYP19A1, CYP1A2, CYP4V2, CYP1B1, CYP24A1, CYP26B1, CYP26C1, CYP2C18, CYP2C9, CYP2U1, CYP39A1, CYP2R1, CYP4Z1, CYP2D7P1 and), while 8 exhibited hypermethylation (CYP51A1, CYP26B1, CYP2R1, CYP2U1, CYP4X1, CYP1A2, CYP2W1, and CYP4V2). Genes such as CYP1A2, CYP26B1, CYP2R1, CYP2U1, and CYP4V2 exhibited both increased and decreased methylation. These genes are crucial for metabolizing eicosanoids, fatty acids, drugs, and diverse substances. Conclusion: The study identified profound methylation changes in multiple CYP genes in the placental tissues relevant to NOWS. This suggests that disruption of DNA methylation patterns in CYP transcripts might play a role in NOWS and may serve as valuable biomarkers, suggesting a future pathway for personalized treatment. Further research is needed to confirm these findings and explore their potential for diagnosis and treatment.

Keywords: CYP1A2; CYP1B1; CYP24A1; CYP4V2; biomarker; cytochromes; neonatal opioid withdrawal syndrome CYP19A1; opioid use.

FIGURE 1

The flow chart illustrates the systematic workflow of genome-wide DNA methylation analysis.

FIGURE 2

Evaluation of the links existing between the evaluated CYP genes. A critical assessment and integration of protein–protein was provided by the STRING database, including direct (physical) and indirect (functional) interactions. String predicted both physical and functional interactions between proteins. Number of nodes: 16, Number of edges: 54, Average node degree: 6.75, Avg. local clustering coefficient: 0.459, Expected number of edges: 1, and PPI enrichment p-value: <0.0001.

FIGURE 3

A detailed Schematic illustration is provided of how epigenetic modifications influence drug metabolism and transport through CYP genes, including specific information about dysregulated CYP enzymes. Additionally, the text outlines the pathogenetic events associated with NOWS and highlights specific CYP genes that undergo significant methylation. Abbreviations. GC: GROUP-SPECIFIC COMPONENT; Ub: ubiquitination; P: phosphorylation; DHCR24: delta (24)-sterol reductase; MARCH6: Membrane Associated Ring-CH-Type Finger 6; PKC: Protein kinase C; PPARG: Peroxisome Proliferator-Activated Receptor Gamma; VDR: Vitamin D Receptor; LXR: Liver X Receptor