AbstractBackgroundMyeloid neoplasms (MN) tend to relapse and deteriorate. Exploring the genomic mutation landscape of MN using next‐generation sequencing (NGS) is a great measure to clarify the mechanism of oncogenesis and progression of MN.MethodsThis multicenter retrospective study investigated 303 patients with MN using NGS from 2019 to 2021. The characteristics of the mutation landscape in the MN subgroups and the clinical value of gene variants were analyzed.ResultsAt least one mutation was detected in 88.11% of the patients (267/303).TET2was the most common mutation in the cohort, followed byGATA2, ASXL1, FLT3, DNMT3A, andTP53. Among patients with myeloid leukemia (ML), multivariate analysis showed that patients aged ≥60 years had lower overall survival (OS,p= 0.004). Further analysis showedTET2, NPM1, SRSF2, andIDH1gene mutations, and epigenetic genes (p< 0.050) presented significantly higher frequency in older patients. In patients with myelodysplastic syndrome (MDS) and myelodysplastic neoplasms (MPN), univariate analysis showed thatBCORL1had a significant impact on OS (p= 0.040); however, in multivariate analysis, there were no factors significantly associated with OS. Differential analysis of genetic mutations showedFLT3, TP53, MUC16, SRSF2, andKDM5Amutated more frequently (p< 0.050) in secondary acute myeloid leukemia (s‐AML) than in MDS and MPN.TP53, U2AF1, SRSF2, andKDM5Awere mutated more frequently (p< 0.050) in s‐AML than in primary AML.KDM5Awas observed to be restricted to patients with s‐AML in this study, and only co‐occurred withMUC16andTP53(2/2, 100%). Another mutation wasMUC16, and its co‐occurrence pattern differed between s‐AML and AML. MUC16 mutations co‐occurred withKDM5AandTP53in 66.7% (2/3) of patients with s‐AML and co‐occurred withCEBPAin 100% (4/4) of patients with AML.ConclusionsOur results demonstrate different genomic mutation patterns in the MN subgroups and highlight the clinical value of genetic variants.