Transient inhibition of translation improves long-term cardiac function after ischemia/reperfusion by attenuating the inflammatory response

Hofmann, C.; Serafin, A.; Schwerdt, O. M.; Younesi, F. S.; Sicklinger, F.; Meyer, I. S.; Malovrh, E.; Sandmann, C.; Juergensen, L.; Kamuf-Schenk, V.; Stroh, C.; Loewenthal, Z.; Rettel, M.; Stein, F.; Katus, H. A.; Jakobi, T.; Frey, N.; Leuschner, F.; Voelkers, M.

doi:10.1101/2022.07.25.501397

bioRxiv · 10.1101/2022.07.25.501397

Transient inhibition of translation improves long-term cardiac function after ischemia/reperfusion by attenuating the inflammatory response

Hofmann, C.·Serafin, A.·Schwerdt, O. M.·Younesi, F. S.·Sicklinger, F.·Meyer, I. S.·Malovrh, E.·Sandmann, C.·Juergensen, L.·Kamuf-Schenk, V.·Stroh, C.·Loewenthal, Z.·Rettel, M.·Stein, F.·Katus, H. A.·Jakobi, T.·Frey, N.·Leuschner, F.·Voelkers, M.

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Abstract

RationaleRapid reperfusion is the most effective treatment for attenuating cardiac injury caused by myocardial ischemia. Yet, reperfusion itself elicits damage to the myocardium through incompletely understood mechanisms, known as ischemia/reperfusion (I/R) injury. The myocardium adapts to I/R by changes in gene expression, which determines the cellular response to reperfusion. Protein translation is a key component of gene expression. However, it is unknown how regulation of translation contributes to cardiac gene expression in response to reperfusion and whether it can be targeted to mitigate I/R injury. MethodsTo examine translation and its impact on gene expression in response to I/R we assessed protein synthesis at different timepoints after ischemia and reperfusion in vitro and in vivo. Pharmacological inhibitors were used to dissect the underlying molecular mechanisms of translational control. Transient inhibition of protein synthesis was undertaken to decipher the effects of the translational response to reperfusion on cardiac function and inflammation. Cell-type-specific ribosome profiling was performed in mice subjected to I/R to determine the impact of translation on the regulation of gene expression in cardiomyocytes. ResultsReperfusion increased translation rates from a previously suppressed state during ischemia in cardiomyocytes, which was associated with the induction of cell death. In vivo, I/R resulted in strong activation of translation in the myocardial border zone. Detailed analysis revealed that the upregulation of translation is mediated by eIF4F complex formation, which was specifically mediated by the mTORC1-4EBP1-eIF4F axis. Short-term pharmacological inhibition of eIF4F complex formation by 4EGI-1 or rapamycin, respectively, attenuated translation, reduced infarct size and improved long-term cardiac function after myocardial infarction. Cardiomyocyte-specific ribosome profiling identified that reperfusion damage increased translation of mRNA networks in cardiomyocytes associated with cardiac inflammation and cell infiltration. Transient inhibition of the mTORC1-4EBP1-eIF4F axis decreased the expression of proinflammatory transcripts such as Ccl2, thereby reducing Ly6Chi monocyte infiltration and myocardial inflammation. ConclusionsMyocardial reperfusion induces protein synthesis in the border zone which contributes to I/R injury by rapidly translating a specific maladaptive mRNA network that mediates immune cell infiltration and inflammation. Transient inhibition of the mTORC1-4EBP1-eIF4F signaling axis during reperfusion attenuates this proinflammatory translational response, protects against I/R injury and improves long-term cardiac function after myocardial infarction. Clinical PerspectiveO_ST_ABSWhat Is New?C_ST_ABSO_LIThis is the first study to investigate the impact of translational regulation on cardiomyocyte gene expression in response to myocardial ischemia/reperfusion. C_LIO_LIWe show that translation regulates approximately two-thirds of differentially expressed genes in cardiomyocytes after ischemia/reperfusion, including many involved in inflammation and immune cell infiltration. C_LIO_LIThe translational response to ischemia/reperfusion is regulated by the mTORC1-4EBP1-eIF4F axis, which determines pro-inflammatory monocyte infiltration via control of the expression of the chemokine Ccl2. C_LI What Are the Clinical Implications?O_LICurrently, there are no specific therapies to prevent ischemia/reperfusion injury, which is mediated, at least in part, by a maladaptive inflammatory response. C_LIO_LIA translationally controlled network regulated by the mTORC1-4EBP1-eIF4F axis can be targeted by a short-term pharmacological intervention to attenuate the inflammatory response and improve cardiac function after ischemia/reperfusion in mice. C_LIO_LIThis study supports the emerging concept of selectively inhibiting maladaptive elements of the inflammatory response to improve outcome in patients after myocardial infarction; in addition, it provides a mechanistic basis for the currently ongoing CLEVER-ACS trial. C_LI

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Hofmann, C., Serafin, A., Schwerdt, O. M., Younesi, F. S., Sicklinger, F., Meyer, I. S., Malovrh, E., Sandmann, C., Juergensen, L., Kamuf-Schenk, V., Stroh, C., Loewenthal, Z., Rettel, M., Stein, F., Katus, H. A., Jakobi, T., Frey, N., Leuschner, F., Voelkers, M.. 2022-07-25. Transient inhibition of translation improves long-term cardiac function after ischemia/reperfusion by attenuating the inflammatory response. https://doi.org/10.1101/2022.07.25.501397

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