S100a9 inhibits Atg9a transcription and participates in suppression of autophagy in cardiomyocytes induced by β1-adrenoceptor autoantibodies

Background: Cardiomyocyte death induced by autophagy inhibition is an important cause of cardiac dysfunction. In-depth exploration of its mechanism may help to improve cardiac dysfunction. In our previous study, we found that beta(1)-adrenergic receptor autoantibodies (beta(1)-AAs) induced a decrease in myocardial autophagy and caused cardiomyocyte death, thus resulting in cardiac dysfunction. Through tandem mass tag (TMT)-based quantitative proteomics, autophagy-related S100a9 protein was found to be significantly upregulated in the myocardial tissue of actively immunized mice. However, whether S100a9 affects the cardiac function in the presence of beta(1)-AAs through autophagy and the specific mechanism are currently unclear. Methods: In this study, the active immunity method was used to establish a beta(1)-AA-induced mouse cardiac dysfunction model, and RT-PCR and western blot were used to detect changes in gene and protein expression in cardiomyocytes. We used siRNA to knockdown S100a9 in cardiomyocytes. An autophagy PCR array was performed to screen differentially expressed autophagy-related genes in cells transfected with S100a9 siRNA and negative control siRNA. Cytoplasmic nuclear separation, co-immunoprecipitation (Co-IP), and immunofluorescence were used to detect the binding of S100a9 and hypoxia inducible factor-1 alpha (HIF-1 alpha). Finally, AAV9-S100a9-RNAi was injected into mice via the tail vein to knockdown S100a9 in cardiomyocytes. Cardiac function was detected via ultrasonography. Results: The results showed that beta(1)-AAs induced S100a9 expression. The PCR array indicated that Atg9a changed significantly in S100a9siRNA cells and that beta(1)-AAs increased the binding of S100a9 and HIF-1 alpha in cytoplasm. Knockdown of S100a9 significantly improved autophagy levels and cardiac dysfunction. Conclusion: Our research showed that beta(1)-AAs increased S100a9 expression in cardiomyocytes and that S100a9 interacted with HIF-1 alpha, which prevented HIF-1 alpha from entering the nucleus normally, thus inhibiting the transcription of Atg9a. This resulted in autophagy inhibition and cardiac dysfunction.

Automated summary

The study found that β(1)-AAs induce the expression of S100a9, leading to cardiomyocyte death and cardiac dysfunction. S100a9 interacts with HIF-1α, inhibiting the transcription of Atg9a and further suppressing autophagy.