Deep proteomics and phosphoproteomics reveal novel biological pathways perturbed by morphine, morphine‐3‐glucuronide and morphine‐6‐glucuronide in human astrocytes

Abstract

Medical use of opioids is essential to treat acute postoperative pain and for different types of chronic pain conditions. However, opioid therapies are associated with complex side effects that are challenging to manage. Long-term use often leads to tolerance, resulting in undertreatment of pain, difficulties estimating a safe dose escalation, and increased risk for overdose. Opioid therapy can also cause opioid-induced hyperalgesia (OIH), a paradoxical painful condition state where patients experience diffused increased sensitivity to pain, often unrelated to the original pain (Lee, Silverman, Hansen, Patel, & Manchikanti, 2011). The opioid epidemic, caused by over-prescription, continues to increase numbers of patients addicted to opioids, as well as users transitioning to heroin and other drugs (Blanco & Volkow, 2019). It is estimated that more than hundred deaths every day in the United States are caused by misuse or opioid overdose (Hser et al., 2019). Although it is generally assumed that physiological homeostatic adaptations cause these side effects, little is known about their underlying molecular and cellular mechanisms.Inside the body, morphine is metabolized into the two main active metabolites morphine-3-glucuronide (M3G) and morphine-6glucuronide (M6G). M6G is found at lower plasma concentration than M3G, but has much higher analgesic potency than M3G ( 200

Tibolone Pre-Treatment Ameliorates the Dysregulation of Protein Translation and Transport Generated by Palmitic Acid-Induced Lipotoxicity in Human Astrocytes: A Label-Free MS-Based Proteomics and Network Analysis

“…We identified 1655 proteins, of which 54 were differentially expressed in pal compared with the control, 128 were regulated in the tibolone 10 nM pre-treatment prior to pal, and 45 in the comparison between the two different treatments. The number of identified non-significant or significant differentiated proteins agrees with other works using human astrocytes [ 52 , 53 , 54 ].…”

Morphine-induced modulation of Nrf2-antioxidant response element signaling pathway in primary human brain microvascular endothelial cells

“…pericytes, astrocytes) are not considered, which may modify morphine transport 23 . Even though morphine concentration in plasma is lower, these concentrations have been used in several studies 1 , 6 , 30 , 69 – 71 in in vitro models and no impact on cytotoxicity or proliferation was denoted in our results, suggesting that they are relevant to investigate dose-dependent effects. Additional in vivo studies would be important to further investigate the effects of morphine on brain microvascular endothelial cells in the context of the neurovascular unit.…”

“…Due to the controversy on morphine relationship with oxidative stress in the current literature and the scarce knowledge available in brain endothelial cells, our aim was to investigate the impact of morphine exposure on human brain endothelial cells. Accordingly, a DIA-proteomics approach was performed in vitro on monocultures of primary HBMECs exposed to morphine for 24 h and 48 h at three different concentrations (1 µM, 10 µM and 100 µM) to explore potential time- and dose-dependent effects 17 , 30 .…”

“…Therefore, we aimed at investigating the impact of morphine exposure on brain endothelial cells via an in vitro monoculture of primary HBMECs and state of the art quantitative mass spectrometry (MS)-based proteomics using a data-independent acquisition (DIA) approach. HBMECs were exposed to morphine at three different concentrations (1, 10 and 100 µM) and for two different time points (24 h and 48 h) to explore dose- and time-dependent effects 17 , 30 . Pathway enrichment analysis revealed alterations in pathways involved in oxidative stress response as well as mitochondrial dysregulations.…”

Tackling the opioid crisis: Novel mechanisms and clinical perspectives

“…In addition to microglia, opioids influence the activity of astrocytes, which are immunocompetent cells within the central nervous system (Dong & Benveniste, 2001;Machelska & Celik, 2020). Dozio et al (2020) apply data-independent acquisition mass spectrometry to delve into how morphine treatment affects the proteome and phosphoproteome of human astrocytes. Bajic et al (2021) then explore central glial reactivity in a model of chemotherapy-induced gut toxicity in the presence of opioid and nonopioid analgesics.…”