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A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
Published on: August 21, 2019
Psilocybin-associated microRNA changes in the pig brain: Establishing an nCounter pipeline for cross-species microRNA
Erik Kaadt1, Rolf Fyllgraf Søkilde1, Hanne Demant Garling2
1Experimental and Molecular Psychiatry, Translational Neuropsychiatry Unit, Department of Clinical Medicine, Aarhus University, Denmark.
Abstract:
Novel treatment strategies are needed to address treatment resistance and delayed onset of action in psychiatry. Psilocybin has shown promise as a treatment for major depressive disorder, but the molecular changes associated with its effects remain incompletely understood. In parallel, microRNAs (miRNAs) have been implicated in depression and in responses to antidepressant interventions. Here, adolescent female pigs were used to explore miRNA expression profiles in the prefrontal cortex (PFC) and hippocampus (HIP) 1 day and 7 days after a single dose of psilocybin. We established a conservative NanoString nCounter workflow for pig tissue using the Human v3b miRNA Assay panel, cross-species reannotation based on complete human-pig sequence conservation, and evaluation of multiple normalization strategies. Using this pipeline, 12 dysregulated miRNAs were identified in the PFC and two in the HIP 1 day after psilocybin administration, whereas four dysregulated miRNAs were detected in the HIP after 7 days. Nine of the 18 miRNAs identified across the two regions have previously been linked to depression-related phenotypes. miR-212-3p and miR-107 showed the most robust acute regulation in the PFC across all four normalization approaches. These findings indicate that psilocybin administration is associated with region- and time-dependent miRNA changes in the pig brain and support the utility of this conservative nCounter pipeline for cross-species miRNA profiling. The study is exploratory and does not establish causality or direct antidepressant mechanisms.
Insights
Psilocybin affects microRNA (miRNA) expression in specific brain regions of pigs, offering insights into potential antidepressant mechanisms. This study highlights region- and time-dependent miRNA changes following psilocybin administration.
Area of Science:
- Neuroscience
- Molecular Psychiatry
- Genomics
Background:
- Novel psychiatric treatments are needed due to resistance and delayed action.
- Psilocybin shows promise for major depressive disorder, but its molecular effects are unclear.
- MicroRNAs (miRNAs) are involved in depression and antidepressant responses.
Purpose of the Study:
- To explore miRNA expression profiles in the prefrontal cortex (PFC) and hippocampus (HIP) after psilocybin administration in adolescent female pigs.
- To investigate region- and time-dependent changes in miRNA expression.
- To validate a cross-species miRNA profiling pipeline.
Main Methods:
- Adolescent female pigs received a single dose of psilocybin.
- miRNA expression was analyzed in PFC and HIP tissues 1 and 7 days post-dose.
- A conservative NanoString nCounter workflow with cross-species reannotation was employed.
- Multiple normalization strategies were evaluated.
Main Results:
- 12 dysregulated miRNAs were identified in the PFC and 2 in the HIP at 1 day post-psilocybin.
- 4 dysregulated miRNAs were detected in the HIP at 7 days post-psilocybin.
- 9 of 18 identified miRNAs are linked to depression-related phenotypes.
- miR-212-3p and miR-107 showed robust acute regulation in the PFC.
Conclusions:
- Psilocybin administration induces region- and time-dependent miRNA changes in the pig brain.
- The developed nCounter pipeline is effective for cross-species miRNA profiling.
- Findings provide preliminary insights into psilocybin's molecular effects, though causality and direct mechanisms are not established.
