# Small molecule FTO inhibitor MO-I-500 protects differentiated SH-SY5Y neuronal cells from oxidative stress

## TL;DR

A small molecule called MO-I-500 protects nerve-like cells from oxidative stress, which could help prevent brain aging and diseases like Alzheimer's.

## Contribution

This study demonstrates that MO-I-500, an FTO inhibitor, protects differentiated SH-SY5Y cells from oxidative stress by reducing ROS and preserving ATP.

## Key findings

- MO-I-500 pretreatment reduces TBHP-induced oxidative damage in dSH-SY5Y cells.
- FTO inhibition shifts cellular metabolism to an energy-efficient state, preserving ATP levels.
- MO-I-500 shows neuroprotective effects in neurons and astrocytes, suggesting dual therapeutic potential.

## Abstract

Oxidative stress is a central driver of brain aging, impairing cellular function and increasing susceptibility to neurodegenerative diseases. Recent studies suggest that the RNA demethylase FTO regulates N6-methyladenosine (m6A) RNA modification, a key pathway in modulating oxidative stress in the brain. However, the precise mechanisms underlying FTO’s role remain unclear. This study examines the neuroprotective potential of MO-I-500, a small-molecule FTO inhibitor, against oxidative stress induced by tert-butyl hydroperoxide (TBHP) in neuron-like SH-SY5Y cells differentiated with retinoic acid and BDNF (dSH-SY5Y).

dSH-SY5Y cells were treated with MO-I-500 alone for 72 h or with TBHP alone for 24 h. Alternatively, cells were pretreated with 1 μM MO-I-500 for 48 h, followed by co-treatment with MO-I-500 and 25 or 50 μM TBHP for an additional 24 h, for a total treatment duration of 72 h. Cellular metabolism was assessed using a Seahorse XF MitoStress assay, and oxidative stress markers, including ROS and superoxide levels, were quantified with DCFDA and MitoSOX probes. ATP content was measured using a bioluminescence assay.

FTO inhibition by MO-I-500 induced a metabolic shift toward an energy-efficient state, enhancing cellular resilience to oxidative stress. Pretreatment significantly reduced TBHP-induced oxidative damage, lowering intracellular ROS levels and preserving ATP content.

Together with our previous findings demonstrating the protective effects of MO-I-500 in astrocytes and recent studies supporting the importance of astrocyte function in neurodegeneration, these results suggest a dual protective role of MO-I-500 in neurons and astrocytes. This dual action positions MO-I-500 as a promising therapeutic strategy to mitigate oxidative damage and reduce the risk of neurodegenerative diseases, including Alzheimer’s disease.

Diagram illustrating the protective effect of the small molecule FTO inhibitor MO-I-500 on dSH-SY5Y neuronal cells from oxidative stress. The left section shows cells under standard conditions with basal energy status. The middle section shows cells under tBHP-induced oxidative stress with increased ROS production. The right section shows the influence of the FTO inhibitor, indicating reduced baseline energy status with maintained ATP production and reduced ROS production. Chemical structure of MO-I-500 is displayed above the arrow pointing toward the effects of the inhibitor.