Acute myeloid leukemia (AML) is a highly heterogeneous hematologic malignancy characterized by dynamic clonal evolution, diverse molecular alterations, and variable therapeutic responses. Although advances in genomic profiling have substantially improved disease classification and risk stratification, many patients continue to experience relapse and treatment resistance, highlighting the need for a more comprehensive understanding of AML biology. Recent developments in multi-omics technologies, including genomics, epigenomics, transcriptomics, proteomics, metabolomics, and single-cell approaches, have enabled unprecedented insights into the molecular mechanisms underlying AML initiation, progression, and therapeutic failure. Integrative multi-omics analyses have revealed complex interactions among genetic mutations, epigenetic remodeling, metabolic reprogramming, and leukemia microenvironmental adaptations that collectively drive disease evolution. These approaches have also facilitated the identification of novel biomarkers, lineage-specific dependencies, and therapeutically actionable vulnerabilities that are not apparent from single-layer analyses. Furthermore, multi-omics-guided studies are accelerating the development of precision medicine strategies by improving patient stratification and uncovering mechanisms of drug resistance. In this review, we summarize recent advances in multi-omics technologies and discuss how integrative analyses are reshaping our understanding of AML evolution. We highlight emerging therapeutic targets identified through multi-omics investigations and examine their translational potential for innovative drug development. Finally, we discuss current challenges and future opportunities for implementing multi-omics-guided precision medicine in AML.