Objective：To explore the genetic alterations and metabolite changes in pediatric acute lymphoblastic leukemia（ALL）. Methods：From September 2021 to May 2022，4 children with ALL who were treated for the first time in the Department of Pediatrics of the First Affiliated Hospital of Nanjing Medical University were selected as the experimental group，and 4 healthy children who underwent physical examination in this hospital at the same time were selected as the control group. Clinical data and serum samples were collected. Serum metabolite profiling was performed by liquid chromatography-mass spectrometry，and genetic variations were analyzed by exome sequencing. Results：Metabolomics revealed that 61 metabolites have difference，with 37 showing increased expression in ALL. The most significant upregulated metabolites were identified as cresol and deoxycholic acid. Additionally，24 metabolites exhibited decreased expression，with 1-pyrroline-5-carboxylic acid and serotonin，etc.，being the most significantly downregulated metabolites. Exonomics has revealed that among the top ten genes ranked by protein interactions include MUC17. Conclusion：There are obvious differences in serum metabolites between children with ALL and healthy children，involving a variety of metabolic pathway changes，and the differential metabolites identified are of great significance for the diagnosis and treatment of ALL. Exome sequencing suggests that genes such as MUC17 may play a certain role in the occurrence of ALL，providing novel concepts and approaches for the study and clinical pharmacotherapy of ALL.