Showing papers by "Massimo Santoro published in 2023"
TL;DR: In this paper , the authors used both in vivo and in vitro approaches to demonstrate that EC possess an endogenous molecular clock and show robust circadian oscillations of core clock genes, which is an internal biological timer that coordinates physiology and gene expression with the 24-hour solar day.
Abstract: AIMS
The circadian clock is an internal biological timer that coordinates physiology and gene expression with the 24-hour solar day. Circadian clock perturbations have been associated to vascular dysfunctions in mammals and a function of the circadian clock in angiogenesis has been suggested. However, the functional role of the circadian clock in endothelial cells (EC) and in the regulation of angiogenesis is widely unexplored.
METHODS AND RESULTS
Here, we used both in vivo and in vitro approaches to demonstrate that EC possess an endogenous molecular clock and show robust circadian oscillations of core clock genes. By impairing the EC-specific function of the circadian clock transcriptional activator BMAL1 in vivo, we detect angiogenesis defects in mouse neonatal vascular tissues, as well as in adult tumor angiogenic settings. We then investigate the function of circadian clock machinery in cultured EC and show evidence that BMAL and CLOCK knock-down impair EC cell cycle progression. By using an RNA- and ChIP-seq genome-wide approaches we identified that BMAL1 binds the promoters of CCNA1 and CDK1 genes and controls their expression in EC.
CONCLUSIONS
Our findings show that EC display a robust circadian clock and that BMAL1 regulates EC physiology in both developmental and pathological contexts. Genetic alteration of BMAL1 can affect angiogenesis in in vivo and in vitro settings.
TRANSLATIONAL PERSPECTIVE
These findings support the need to explore the manipulation of the circadian clock in vascular diseases. Further investigation of the behavior of BMAL1 and its target genes in the tumor endothelium can aim to discover novel therapeutic interventions to interfere with the endothelial circadian clock in the tumor context.
1 citations
TL;DR: Oberkersch et al. as mentioned in this paper presented a protocol for evaluating postnatal angiogenesis in a mouse hind limb ischemia model, which induces a severe restriction of blood supply of the femoral artery and vein.
Abstract: Hind limb ischemia is a useful model to assess metabolic and cellular responses. Here, we present a protocol for evaluating post-natal angiogenesis in a mouse hind limb ischemia model. We describe steps to induce a severe restriction of blood supply of the femoral artery and vein that mimics the real-life scenario observed in clinical settings. We then detail procedures for follow-up laser Doppler imaging to compare post-ischemic responses of four different mouse strains in their capacity to trigger compensatory arteriogenesis. For complete details on the use and execution of this protocol, please refer to Oberkersch et al. (2022).1.