HuR inhibition reduces post-ischemic cardiac remodeling by dampening acute inflammatory gene expression and the innate immune response

Slone, S.; Anthony, S. R.; Green, L. C.; Nieman, M. L.; Alam, P.; Wu, X.; Roy, S.; Aube, J.; Xu, L.; Lorenz, J. N.; Owens, A. P.; Kanisicak, O.; Tranter, M.

doi:10.1101/2023.01.17.524420

bioRxiv · 10.1101/2023.01.17.524420

HuR inhibition reduces post-ischemic cardiac remodeling by dampening acute inflammatory gene expression and the innate immune response

Slone, S.·Anthony, S. R.·Green, L. C.·Nieman, M. L.·Alam, P.·Wu, X.·Roy, S.·Aube, J.·Xu, L.·Lorenz, J. N.·Owens, A. P.·Kanisicak, O.·Tranter, M.

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Abstract

Myocardial ischemia/reperfusion (I/R) injury and the resulting cardiac remodeling is a common cause of heart failure. The RNA binding protein Human Antigen R (HuR) has been previously shown to reduce cardiac remodeling following both I/R and cardiac pressure overload, but the full extent of the HuR-dependent mechanisms within cells of the myocardium have yet to be elucidated. In this study, we applied a novel small molecule inhibitor of HuR to define the functional role of HuR in the acute response to I/R injury and gain a better understanding of the HuR-dependent mechanisms during post-ischemic myocardial remodeling. Our results show an early (two hours post-I/R) increase in HuR activity that is necessary for early inflammatory gene expression by cardiomyocytes in response to I/R. Surprisingly, despite the reductions in early inflammatory gene expression at two hours post-I/R, HuR inhibition has no effect on initial infarct size at 24-hours post-I/R. However, in agreement with previously published work, we do see a reduction in pathological remodeling and preserved cardiac function at two weeks post-I/R upon HuR inhibition. RNA-sequencing analysis of neonatal rat ventricular myocytes (NRVMs) at two hours post-LPS treatment to model damage associated molecular pattern (DAMP)-mediated activation of toll like receptors (TLRs) demonstrates a broad HuR-dependent regulation of pro-inflammatory chemokine and cytokine gene expression in cardiomyocytes. We show that conditioned media from NRVMs pre-treated with HuR inhibitor loses the ability to induce inflammatory gene expression in bone marrow derived macrophages (BMDMs) compared to NRVMs treated with LPS alone. Functionally, HuR inhibition in NRVMs also reduces their ability to induce endocrine migration of peripheral blood monocytes in vitro and reduces post-ischemic macrophage infiltration to the heart in vivo. In summary, these results suggest a HuR-dependent expression of pro-inflammatory gene expression by cardiomyocytes that leads to subsequent monocyte recruitment and macrophage activation in the post-ischemic myocardium.

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Slone, S., Anthony, S. R., Green, L. C., Nieman, M. L., Alam, P., Wu, X., Roy, S., Aube, J., Xu, L., Lorenz, J. N., Owens, A. P., Kanisicak, O., Tranter, M.. 2023-01-18. HuR inhibition reduces post-ischemic cardiac remodeling by dampening acute inflammatory gene expression and the innate immune response. https://doi.org/10.1101/2023.01.17.524420

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