Exposure to Ketamine and 2-Fluorodeschloroketamine ImpairsMitochondrial Oxidative Phosphorylation in Human Cerebral Organoids:Implications for Neurodevelopmental Toxicity

INTRODUCTION: Ketamine and its structural analog, 2-Fluorodeschloroketamine, both potent stimulants, can induce euphoria but also cause neurotoxicity, cognitive decline, and neurodevelopmental deficits in the fetus. However, the molecular mechanisms responsible for these neurodevelopmental abnormalities are not yet fully elucidated, particularly concerning effects that are specific to certain cell types. METHODS: Human cerebral organoids were used as a model, and single-cell transcriptomics was conducted to evaluate the effects of prenatal exposure to KET or 2-FDCK (30 μM) on fetal brain development. A total of 83,436 cells from both control and treated organoids were analyzed. Key findings were corroborated through the assessment of mitochondrial dysfunction and bioenergetic deficiencies following KET or 2-FDCK exposure in primary cortical neurons isolated from fetal mice. RESULTS: The analysis revealed that the cerebral organoids contained a diverse range of glial and neuronal cell types. Importantly, the findings demonstrated that both substances induced corticalspecific gene expression networks involved in the regulation of mitochondrial oxidative phosphorylation. The results indicated that KET and 2-FDCK exposure can augment mitochondrial fragmentation and oxidative stress, accompanied by a significant decrease in ATP production capacity, thereby increasing the risk of the fetus to neurological diseases through neurodevelopmental damage. DISCUSSION: Disruptions in energy production during rapid developmental periods can make the offspring more vulnerable to neurological issues. The research highlights how cerebral organoids serve as a valuable tool for studying how substance exposures affect brain development, thereby enhancing the understanding of these important effects. CONCLUSION: In conclusion, the results offer direct evidence of the neurodevelopmental toxicity associated with KET and 2-FDCK following prenatal exposure, utilizing cerebral organoids as an invaluable translational model. It was recognized that mitochondrial oxidative phosphorylation disruption was a probable primary molecular mechanism. These findings highlight the substantial risks these substances pose to fetal brain development.