Drug dependence remains one of the most persistent threats to public health, causing serious negative consequences for individuals, their families, and society (Lo et al., 2020; Heilig et al., 2021). The euphoria produced by addictive substances, including methamphetamine (METH), is often linked to drug paraphernalia or environments associated with prior use. Re-exposure to these cues activates the nucleus accumbens (NAc) a key component of the brain's reward circuitry and increases the likelihood of repeated drug intake (Zhang et al., 2023; Li et al., 2024; D'souza et al., 2021; Scheyer et al., 2016; LeGates et al., 2018). This, in turn, contributes to addiction-related neural plasticity (Tu et al., 2019; Hedges et al., 2018; Simmler et al., 2022). Specifically, the NAc plays a critical role in facilitating psychostimulant dependence; for example, drug-seeking behaviors following withdrawal are supported by NAc activity (Ren et al., 2015; West and Carelli, 2016). The psychostimulant effects of METH addiction are associated with elevated extracellular dopamine (DA) levels within the NAc (Bhimani et al., 2021; Yorgason et al., 2020). Behavioral paradigms such as conditioned place preference (CPP) have demonstrated that dopamine D1 and D2 receptor antagonists administered in the NAc attenuate the incentive properties of drug rewards (Farzinpour et al., 2018; Kawahara et al., 2021). Therefore, it has been proposed that the dopaminergic system in the NAc plays a vital role in consolidating cue-related reward effects and mediating drug-induced CPP (O'Neal et al., 2022; Wu et al., 2024).

Clinically, dysfunction of mRNA N4-acetylcytidine (ac4C) results in a syndromic form of intellectual disability characterized by behavioral abnormalities, hearing loss, and facial dysmorphism (Broly et al., 2022). N-acetyltransferase 10 (NAT10), a member of the Gcn5-related N-acetyltransferases (GNAT) family, is a nucleolar protein that catalyzes ac4C modification. Upregulation of NAT10 leads to elevated ac4C expression, which exacerbates seizure behavior (Zhang et al., 2025). Previous studies have shown that the NAT10 motif contains an acetyl-CoA binding region, a tRNA binding region, and an N-acetyltransferase structural domain, suggesting a physiological role for NAT10 in regulating oxidative stress and metabolism across multiple cancer types (Tao et al., 2021; Dalhat et al., 2021). Additionally, RNA helicase and NAT domains are essential for primary tumor growth and brain metastasis in vivo. NAT10 promotes the expression of 3-phosphoglycerate dehydrogenase and phosphoserine aminotransferase 1 two enzymes involved in serine biosynthesis and implicated in brain metastasis supporting the role of NAT10 as a key regulator of brain metastasis (Chen et al., 2025).

Recent studies increasingly demonstrate that NAT10 plays a significant role in the treatment of neuropsychological disorders such as depression and epilepsy, thus holding important clinical significance. 4-(4-Cyanophenyl)-2-(2-cyclopentylidenehydrazinyl) thiazole, also known as Remodelin, which primarily functions in clinical areas such as cancer suppression and adjuvant anti-cancer therapy (Luo et al., 2025; Jin et al., 2022), a recent study indicated that Remodelin as effective inhibitor of NAT10 prevents seizure behavior induced by intrahippocampal kainic acid injection, which highlighted the novel regulatory role of NAT10-mediated ac4C modification in neuropsychiatric conditions (Zhang et al., 2025). Additionally, exposure to chronic mild stress leads to NAT10 upregulation in the mouse hippocampus, while neuron-specific overexpression of NAT10 in the hippocampus induces anxiety- and depression-like behaviors (Guo et al., 2022). Moreover, microglial activation induced by encephalopathy and the subsequent release of inflammatory mediators increase NAT10 expression in neurons, which in turn promotes GABABR1 expression through mRNA acetylation, ultimately resulting in cognitive dysfunction (Gao et al., 2024). Notably, other members of the GNAT family such as arylalkylamine N-acetyltransferase (AANAT), Shati/N-acetyltransferase-8-like protein (NAT8L), and α-glucosaminide N-acetyltransferase (HGSNAT) also contribute to learning and memory impairment (Ghorbandaiepour et al., 2024; Haddar et al., 2021; Pan et al., 2022). For instance, NAT8L is highly expressed in the NAc of mice repeatedly exposed to METH, serving as a homeostatic mechanism that mitigates CPP and hyperlocomotion by enhancing both plasmalemmal and vesicular dopamine (DA) uptake and reducing METH-induced DA overflow in the NAc (Haddar et al., 2019; Uno et al., 2017). Collectively, these findings suggest that the GNAT family plays a role in both cognition and memory. Although the specific function of NAT10 in addiction remains unclear, we hypothesize based on currently available evidence that NAT10 plays a critical role in addiction-related mechanisms.

In this study, two METH-induced addiction models CPP and hyperlocomotion were established by intraperitoneal injection of 2 mg/kg METH into C57BL/6J mice. We observed a significant increase in neuronal NAT10 expression in the NAc of METH-treated mice, accompanied by pronounced addiction-related behaviors. Pharmacological antagonism of dopamine D1 receptors (D1R) attenuated METH-induced hyperlocomotion and normalized NAT10 expression levels. Furthermore, neuronal inhibition of NAT10 in the NAc reduced PSD95 protein levels, decreased dendritic spine density, and subsequently suppressed METH-induced CPP and hyperlocomotion. Collectively, these results indicate that NAT10 expression in the NAc contributes to METH-induced hyperlocomotion and CPP, mediated by dopaminergic signaling and synaptic plasticity. These findings suggest that NAT10 may serve as a novel target for the prevention and treatment of METH-induced addiction.