Integration of Molecular Inflammatory Interactome Analyses Reveals Dynamics of Circulating Cytokines and Extracellular Vesicle Long Non-Coding RNAs and mRNAs in Heroin Addicts During Acute and Protracted Withdrawal

Abstract

Heroin addiction and withdrawal influence multiple physiological functions, including immune responses, but the mechanism remains largely elusive. The objective of this study was to investigate the molecular inflammatory interactome, particularly the cytokines and transcriptome regulatory network in heroin addicts undergoing withdrawal, compared to healthy controls (HCs). Twenty-seven cytokines were simultaneously assessed in 41 heroin addicts, including 20 at the acute withdrawal (AW) stage and 21 at the protracted withdrawal (PW) stage, and 38 age- and gender-matched HCs. Disturbed T-helper(Th)1/Th2, Th1/Th17, and Th2/Th17 balances, characterized by reduced interleukin (IL)-2, elevated IL-4, IL-10, and IL-17A, but normal TNF-α, were present in the AW subjects. These imbalances were mostly restored to the baseline at the PW stage. However, the cytokines TNF-α, IL-2, IL-7, IL-10, and IL-17A remained dysregulated. This study also profiled exosomal long non-coding RNA (lncRNA) and mRNA in the plasma of heroin addicts, constructed co-expression gene regulation networks, and identified lncRNA-mRNA-pathway pairs specifically associated with alterations in cytokine profiles and Th1/Th2/Th17 imbalances. Altogether, a large amount of cytokine and exosomal lncRNA/mRNA expression profiling data relating to heroin withdrawal was obtained, providing a useful experimental and theoretical basis for further understanding of the pathogenic mechanisms of withdrawal symptoms in heroin addicts.

Bioinformatic Tools for the Analysis and Prediction of ncRNA Interactions

“…Noncoding RNA (ncRNA) genes are those genes that produce transcripts or functional RNAs that, unlike messenger RNAs, are not translated into proteins. Only a few years ago these transcripts were considered “dark matter” in the genome, but now they play leading roles in the regulation of biological processes [ 33 , 38 ].…”

“…Our knowledge of the roles of ncRNAs in biological processes that impact human health has largely been obtained from experimental findings [ 31 , 32 , 33 , 34 ]. However, computational biology can significantly boost this knowledge, providing insights into the possible interactions of these ncRNAs with other molecules, which can subsequently be verified experimentally [ 35 , 36 , 37 ].…”

“…However, computational biology can significantly boost this knowledge, providing insights into the possible interactions of these ncRNAs with other molecules, which can subsequently be verified experimentally [ 35 , 36 , 37 ]. Hence, computational predictions are an alternative approach that allows the expansion of knowledge through a process of mutual enrichment; this is because the information that is obtained experimentally continuously feeds into computational methods, and, in turn, the results of these predictions suggest possible interactions that can subsequently be validated through laboratory experiments using in vitro and/or in vivo models [ 31 , 33 , 34 ].…”

Whole Transcriptome Sequencing of Peripheral Blood Shows That Immunity/GnRH/PI3K-Akt Pathways Are Associated With Opioid Use Disorder

“…Multiple brain regions, such as the nucleus accumbens (NAc), central nucleus of the amygdala (CeA) and prefrontal cortex (PFC), as well as the peripheral system, have been reported to be associated with addiction (3). Previous studies have revealed that metabolism, endocrine systems, immune systems and mitochondria play important roles in the process of long-lived behavioral abnormalities associated with addiction (4)(5)(6)(7)(8). Opiate addiction leads to autophagy-mediated dysfunction of mitochondria, such as a decrease in mitochondrial DNA (mtDNA) copy number in the hippocampus and peripheral blood and an increase in reactive oxygen species (ROS), which contributes to cell damage and apoptosis (6).…”

“…Opiate addiction leads to autophagy-mediated dysfunction of mitochondria, such as a decrease in mitochondrial DNA (mtDNA) copy number in the hippocampus and peripheral blood and an increase in reactive oxygen species (ROS), which contributes to cell damage and apoptosis (6). In addition, recent studies have suggested that many cytokines and regulatory T cells in peripheral blood were dysregulated in patients with heroin addiction compared to healthy controls (8)(9)(10)(11). Women with addiction had lower oxytocin levels in their peripheral blood, which could be used as a biomarker for predicting the intensity of social anxiety in female patients with heroin withdrawal (12).…”