Inhibition of miR-20a promotes neural stem cell survival under oxidative stress conditions

Ivan Arzhanov; Ruslan A Klassen; Lukas Valihrach; Nataliya Romanyuk

doi:10.3389/fnins.2025.1601101

Inhibition of miR-20a promotes neural stem cell survival under oxidative stress conditions

Front Neurosci. 2025 Jun 26:19:1601101. doi: 10.3389/fnins.2025.1601101. eCollection 2025.

Authors

Ivan Arzhanov^{1

2}, Ruslan A Klassen^{3

4}, Lukas Valihrach³, Nataliya Romanyuk¹

Affiliations

¹ Department of Neuroregeneration, Institute of Experimental Medicine CAS, Prague, Czechia.
² Department of Neuroscience, 2nd Medical Faculty, Charles University, Prague, Czechia.
³ Laboratory of Glial Biology and Omics Technologies, Institute of Biotechnology of the Czech Academy of Sciences - BIOCEV, Vestec, Czechia.
⁴ Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, UCT Prague, Prague, Czechia.

Abstract

Introduction: Oxidative stress (OS) is a key contributor to secondary damage following spinal cord injury (SCI), leading to neural stem cell (NSC) dysfunction and apoptosis. MicroRNA-20a (miR-20a) is upregulated after SCI and plays a role in regulating apoptosis and survival pathways. This study explores the therapeutic potential of miR-20a inhibition in mitigating OS-induced damage in NSCs.

Methods: Human iPSC-derived NSCs were subjected to oxidative stress by exposure to 100 µM hydrogen peroxide (H₂O₂) for 2 hours, followed by treatment with a miR-20a inhibitor (100 nM) to attenuate the adverse effects. Metabolic activity was evaluated using the Alamar Blue assay. Apoptotic responses and miR-20a expression levels were assessed via flow cytometry, RT-qPCR, and Western blot analysis.

Results: NSCs exposed to OS showed a marked reduction in metabolic activity. However, treatment with a miR-20a inhibitor over 72 h significantly improved cell survival and metabolic activity in a time-dependent manner compared to untreated stressed cells.

Discussion: Our findings suggest that miR-20a inhibition mitigates OS-induced cytotoxicity and promotes NSC viability, presenting a potential therapeutic approach for enhancing neural tissue regeneration.

Keywords: microRNA-20a; neural apoptosis; neural stem cells; neuroprotection; oxidative stress.