Cardiomyocytes stimulate angiogenesis after ischemic injury in a ZEB2-dependent manner

The disruption in blood supply due to myocardial infarction is a critical determinant for infarct size and subsequent deterioration in function. The identification of factors that enhance cardiac repair by the restoration of the vascular network is, therefore, of great significance. Here, we show that the transcription factor Zinc finger E-box-binding homeobox 2 (ZEB2) is increased in stressed cardiomyocytes and induces a cardioprotective cross-talk between cardiomyocytes and endothelial cells to enhance angiogenesis after ischemia. Single-cell sequencing indicates ZEB2 to be enriched in injured cardiomyocytes. Cardiomyocyte-specific deletion of ZEB2 results in impaired cardiac contractility and infarct healing post-myocardial infarction (post-MI), while cardiomyocyte-specific ZEB2 overexpression improves cardiomyocyte survival and cardiac function. We identified Thymosin β4 (TMSB4) and Prothymosin α (PTMA) as main paracrine factors released from cardiomyocytes to stimulate angiogenesis by enhancing endothelial cell migration, and whose regulation is validated in our in vivo models. Therapeutic delivery of ZEB2 to cardiomyocytes in the infarcted heart induces the expression of TMSB4 and PTMA, which enhances angiogenesis and prevents cardiac dysfunction. These findings reveal ZEB2 as a beneficial factor during ischemic injury, which may hold promise for the identification of new therapies.


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Sampling strategy 2 For all animals experiments, 6 sham and 12 MI samples were used according to standard scientific conventions, as indicated in each figure.The sample size was determined by a power calculation based upon an echocardiographic effect size.For baseline animal studies and in vitro studies estimates were made based on our previous experience, experimental approach,availability and feasibility required to to obtain statistically significant results.
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We used the HUVECs that were obtained from Lonza #CC-2519 and NIH-3T3 cell line obtained from Sigma-Aldrich #86052701, Immortalized rat neonatal heart derived cells H10 cells (Jahn et al. Journal of Cell Science 1996) Cells were authenticated by examination of morphology and gene expression.Also, all cell lines were carefully labeled and stored until use.
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