ZBP1 exacerbates myocardial ischemia-reperfusion injury through the induction of non-canonical cardiomyocyte PANoptosis

Abstract Background Myocardial ischemia-reperfusion (I/R) injury is the primary factor that counteracts the beneficial effects of reperfusion therapy. Cardiomyocyte death serves as the fundamental pathological hallmark of I/R injury. However, targeting a single type of cell death has been reported as ineffective for preventing I/R injury. Z-DNA-binding protein 1 (ZBP1) is well-established as a nucleic acid sensor that activates inflammatory and various cell death signaling pathways. However, the specific role of ZBP1 in adult cardiomyocytes, particularly in the absence of nucleic acid ligands, remains largely unexplored. Methods RNA-sequencing analysis was used to identify critical factors involved in myocardial I/R injury. The role of ZBP1 in cardiomyocytes was investigated using an I/R-injury model with cardiomyocyte-specific, conditional Zbp1-knockout (Zbp1fl/flMyh6+) and -overexpression (Zbp1TGMyh6+) mice. The mechanism of ZBP1-regulated cardiomyocyte death was further explored through the application of cell-death inhibitors. Results Dynamic transcriptomic analyses at various I/R stages identified a cluster of genes significantly enriched in cell death-related processes, with ZBP1 showing significant expression changes in both our I/R-injury mouse model and human ischemic cardiomyopathy datasets. Cardiomyocytes were the primary cell type expressing ZBP1 in response to I/R Injury. Hypoxia/reoxygenation stress induced the upregulation of multiple cell-death markers indicative of PANoptosis in adult cardiomyocytes, which was mitigated by ZBP1 deficiency. Cardiomyocyte-specific Zbp1 knockout ameliorated I/R-induced PANoptosis, resulting in a more substantial reduction in myocardial infarct size compared to treatment with conventional cell-death inhibitors. Conversely, myocardial Zbp1 overexpression in adult mice directly induced cardiac remodeling and heart failure. Mechanistically, ZBP1 drives cardiomyocyte PANoptosis by facilitating the formation of the ZBP1/RIPK3/CASP8/CASP6 PANoptosome complex, while certain canonical components, including RIPK1, NLRP3, and ASC, are not essential for this process. Conclusions These findings highlight the role of ZBP1 as a mediator of cardiomyocyte non-canonical PANoptosis and suggest that developing effective agents which target ZBP1 could be a promising strategy for mitigating myocardial I/R injury.Zbp1 knockout ameliorated I/R injury Zbp1 overexpression induced HF