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Morphine-Driven m6A Epitranscriptomic Neuroadaptations in Primary Cortical Cultures.

Konrad R Dabrowski^1,2, Stephanie E Daws^3,4

¹Center for Substance Abuse Research, Temple University, Philadelphia, PA, USA.

Molecular Neurobiology

|May 23, 2024

Summary

Opioid use alters RNA methylation (m6A) in the brain, potentially through the AlkB Homolog 5 (Alkbh5) enzyme. This study reveals morphine-induced epitranscriptomic changes, offering new insights into opioid use disorder mechanisms.

Keywords:

Alkbh5 Morphine N6-methyladenosine Neurons Opioids RNA m6A

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Area of Science:

Neuroscience
Molecular Biology
Epigenetics

Background:

Opioid overdose is a leading cause of accidental death, necessitating research into neurobiological consequences.
While opioid effects on the transcriptome and epigenome are known, epitranscriptomic regulation via RNA modifications is understudied.
N6-methyladenosine (m6A) RNA methylation is prevalent in the brain and crucial for cognitive functions, yet its role in opioid use disorder remains unexplored.

Purpose of the Study:

To investigate the role of m6A RNA modifications in the context of opioid use disorder.
To determine if chronic opioid exposure, specifically morphine, alters m6A-modifying enzymes in the brain.
To test the hypothesis that morphine regulates m6A modifications through modulation of the m6A demethylase AlkB Homolog 5 (Alkbh5).

Main Methods:

Primary rat cortical cultures were treated with morphine to assess changes in m6A-modifying enzymes.
Alkylation Homolog 5 (Alkbh5) enzyme activity was modulated via knockdown.
m6A RNA modification profiles were compared between morphine-treated cultures and those with Alkbh5 knockdown.

Main Results:

Morphine treatment significantly regulated m6A-modifying enzymes, including Alkbh5, in cortical cultures.
Morphine treatment and Alkbh5 knockdown resulted in concordant m6A epitranscriptomic profiles for a subset of transcripts.
Commonly regulated transcripts were associated with key neuronal functions such as serotonin secretion, synapse disassembly, neuron remodeling, and immune response.

Conclusions:

Morphine exposure induces significant epitranscriptomic changes in the brain.
A portion of these morphine-driven m6A modifications may be mediated by the downregulation of Alkbh5.
These findings highlight the potential role of epitranscriptomic alterations in opioid use disorder and suggest Alkbh5 as a target for future research.