Background: Stuttering is a neurodevelopmental disorder of unclear etiology characterized by involuntary disruptions in speech fluency. Recently, stuttering has been shown to be related to a point mutation in GNPTAB, a housekeeping gene involved in cellular lysosomal enzyme-targeting pathway, though it remained unclear how such a mutation might cause the stuttering phenotype. Transcranial ultrasounds in adults who stutter detected mesencephalic hyperechogenicity, likely associated with iron deposits. The current study aims to confirm, assess, and alleviate iron accumulation in a transgenic mouse model engineered with a mutation associated with Gnptab. Methods: Animal studies: All procedures in this study were approved by the Animal Care and Use Committee of the Intramural Research Program of the National Institute of Neurological Disorders and Stroke. Vocalization studies: Ultrasonic vocalizations (USVs) in male mice were recorded via Avisoft Bioacoustics and SASLab Pro software, and the complexity and geometry of USVs were analyzed in Sonotrack (Metris Inc). Iron chelation therapy: Adult male Gnptab-mutant (n = 10) and control littermates (n = 10) were treated by 3- Hydroxy-1,2-dimethyl-4(1H)-pyridone (Deferiprone; 0.1 g/L) orally for 1 month via water supply. Animals' vocal behaviors were recorded and compared before and after the treatment. Histological processing: Mice were transcardially perfused with 4% paraformaldehyde (PFA) fixative. The brain was extracted and post-fixed for 3-5 days in PFA. Iron was detected by iron-sensitive DAB-enhanced Perl’s staining on coronal brain sections. Bright field images were taken and imported to Fiji, where regional iron content was quantified. Statistical analysis: All data was exported to Prism 9.0. Statistical significance was taken at p < 0.05. Results: Gnptab-mutant animals show higher percentage of complex USVs and higher iron deposition in the medial lateral striatum, dorsolateral striatum, and central striatum when analyzed for percent area of iron deposition following Perl’s staining. Alleviation of this iron accumulation via chelation therapy was confirmed by Perl’s staining. Strikingly, the percentage of complex vocal bouts decreased following iron chelator therapy. Conclusion: The mutation in Gnptab gene, associated with stuttering, leads to an accumulation of CNS iron that may be alleviated with iron chelation, ultimately improving the stuttering phenotype in mice. Accordingly, we posit a novel therapeutic option and long-awaited etiology of the disorder.