Purpose <p>Adamantinomatous craniopharyngioma (ACP) are rare and histologically benign but clinically challenging neoplasms. The aim of this study was to investigate the molecular genetic and proteomic alterations in the tumor to identify potential therapeutic targets for ACP.</p> Methods <p>Sanger sequencing characterized mutations in 20 craniopharyngiomas. Serum proteomics by iTRAQ-based LC-MS/MS compared ACP patients with healthy controls. KNG1 expression was examined in tumor tissues (RT‑qPCR, Western blot) and serum (ELISA), followed by clinical correlation analysis. Molecular mechanisms of KNG1 were investigated using snRNA‑seq, spatial transcriptomics (ST), and immunofluorescence. To investigate whether KNG1 functions through the PI3K-AKT pathway, we performed lentiviral overexpression in ACP primary cells and applied a PI3K-AKT activator. The resulting impact on proliferation, migration, invasion, and mitochondrial integrity was then examined using a suite of functional assays.</p> Results <p>CTNNB1 missense mutations were identified in 38.8% of ACPs. Subsequently, KNG1 was found downregulated at both tissue and serum levels, with the latter inversely correlating with tumor volume. Mechanistically, snRNA-seq, ST and immunofluorescence analyses suggested the involvement of the PI3K-AKT pathway. Functional studies confirmed that KNG1 overexpression attenuated PI3K-AKT signaling, curbed tumor growth, migration and invasion, and promoted mitochondrial damage, all of which were rescued by PI3K-AKT activation.</p> Conclusion <p>Our findings indicate that downregulation of KNG1 is associated with ACP progression, potentially through its interaction with PI3K-AKT pathway activation. These results support the exploration of KNG1 and PI3K-AKT signaling as potential therapeutic targets in ACP, and suggest that serum KNG1 may serve as a candidate biomarker for monitoring disease progression.</p>