Abstract 33: Cardiac Fibroblasts Reprogram Into Endothelial Like Cells in a p53 Dependent Manner After Acute Ischemic Cardiac Injury

Abstract

The mammalian heart exhibits poor regenerative capacity after acute ischemic injury and heals primarily by fibrosis. Recently, several groups have demonstrated that cardiac fibroblasts can be reprogrammed to adopt myogenic fates using exogenous transcription factors. However, the ability of cardiac fibroblasts to adopt specific cellular fates in the absence of exogenous factors is unclear. Here, we demonstrate that a subset of cardiac fibroblasts adopt endothelial characteristics after ischemic cardiac injury in the absence of any added factors. Using mice harboring genetically labeled fibroblasts (Col1a2CreERT:R26RTdTomato), we show that 34 +/- 3% (mean, SEM) of labeled cardiac fibroblasts in the injury border zone express endothelial markers such as VE-cadherin. Fibroblast derived endothelial cells comprised 25 +/- 2% of total and 8 +/- 2% of luminal endothelial cells at the border zone 3 days after injury. To better understand fibroblast-endothelial reprogramming we subjected cardiac fibroblasts to cellular stress (serum starvation) and found that they formed tubes on Matrigel and up-regulated endothelial specific genes (e.g. VE-cadherin, Flk1, Flt1) 6-20 fold. We show that reprogramming of fibroblasts to endothelial like cells ex vivo is p53 dependent. Inhibiting p53 activity by pharmacological means (Pifithrin-α) or genetic deletion in fibroblasts (Col1a2CreERT:p53fl/fl) led to a 94% decrease in Matrigel tube formation and 90% reduction in endothelial gene expression. Moreover, we observed that p53 levels in cardiac fibroblasts were more than 10-fold higher at the injury border zone using semi-quantitative immunofluorescent staining. Injection of a p53 activator after injury doubled p53 levels in cardiac fibroblasts and increased the rate of fibroblast-endothelial reprogramming by 43%. Enhanced fibroblast-endothelial reprogramming was also associated with decreased collagen deposition 3 days post injury. In summary, we show that cardiac fibroblasts are able to adopt endothelial cell like fates both in vivo and ex vivo in a p53 dependent manner. Manipulation of fibroblast to endothelial reprogramming could represent a novel therapeutic strategy to increase post infarct angiogenesis and enhance function in the injured heart.