PANoptosis represents a distinct form of programmed cell death (PCD) that is characterized by the integration of key features from apoptosis, pyroptosis, and necroptosis but cannot be fully explained by any single cell death pathway. A key trigger of PANoptosis is the assembly and activation of the PANoptosome. To date, four types of PANoptosomes have been identified, including PANoptosomes nucleated by Z-DNA binding protein 1 (ZBP1), absent in melanoma 2 (AIM2), receptor-interacting protein kinase 1 (RIPK1), and NOD-like receptor protein 12 (NLRP12). This review summarizes current advances in the molecular regulatory mechanisms underlying PANoptosis and delineates the assembly and activation processes of several PANoptosome complexes. Furthermore, the role of PANoptosis in various pathologic conditions, such as infectious diseases, autoimmune disorders, neurodegenerative diseases, and cancer, are discussed. Recent preclinical and clinical advances in targeting the PANoptosis pathway through small molecule inhibitors, nanomedicines, and combination therapies for the treatment of infectious diseases, inflammatory conditions, and malignancies are also highlighted. However, current research involving the mechanisms underlying PANoptosis are preliminary and the clinical translation faces significant challenges, necessitating further in-depth exploration. This review demonstrates that targeting molecular components and signaling pathways of PANoptosis represents a highly promising therapeutic strategy, aiming to provide a theoretical foundation for the development of novel clinical treatments for related diseases.