Diabetes Impairs Angiogenesis and Induces Endothelial Cell Senescence by Up-Regulating Thrombospondin-CD47-Dependent Signaling

Abstract

Endothelial dysfunction, impaired angiogenesis and cellular senescence in type 2 diabetes constitute dominant risk factors for chronic non-healing wounds and other cardiovascular disorders. Studying these phenomena in the context of diabetes and the TSP1-CD-47 signaling dictated the use of the in vitro wound endothelial cultured system and an in vivo PVA sponge model of angiogenesis. Herein we report that diabetes impaired the in vivo sponge angiogenic capacity by decreasing cell proliferation, fibrovascular invasion and capillary density. In contrast, a heightened state of oxidative stress and elevated expression of TSP1 and CD47 both at the mRNA and protein levels were evident in this diabetic sponge model of wound healing. An in vitro culturing system involving wound endothelial cells confirmed the increase in ROS generation and the up-regulation of TSP1-CD47 signaling as a function of diabetes. We also provided evidence that diabetic wound endothelial cells (W-ECs) exhibited a characteristic feature that is consistent with cellular senescence. Indeed, enhanced SA-β-gal activity, cell cycle arrest, increased cell cycle inhibitors (CKIs) p53, p21 and p16 and decreased cell cycle promoters including Cyclin D1 and CDK4/6 were all demonstrated in these cells. The functional consequence of this cascade of events was illustrated by a marked reduction in diabetic endothelial cell proliferation, migration and tube formation. A genetic-based strategy in diabetic W-ECs using CD47 siRNA significantly ameliorated in these cells the excessiveness in oxidative stress, attenuation in angiogenic potential and more importantly the inhibition in cell cycle progression and its companion cellular senescence. To this end, the current data provide evidence linking the overexpression of TSP1-CD47 signaling in diabetes to a number of parameters associated with endothelial dysfunction including impaired angiogenesis, cellular senescence and a heightened state of oxidative stress. Moreover, it may also point to TSP1-CD47 as a potential therapeutic target in the treatment of the aforementioned pathologies.

SDF-1α Gene-Activated Collagen Scaffold Restores Pro-Angiogenic Wound Healing Features in Human Diabetic Adipose-Derived Stem Cells

“…One of the significant challenges in the healing of diabetic wounds is the lack of angiogenesis [ 72 ]. Therefore, having noted an enhanced pro-angiogenic profile in transfected diabetic ADSCs, we assessed the pro-angiogenic bioactivity of the secreted factors on endothelial cells.…”

Focused shockwave therapy in diabetic foot ulcers: secondary endpoints of two multicentre randomised controlled trials

“…26 Endothelial cells explanted from DFUs into in vitro sponge models of healing demonstrated impaired angiogenesis and expression of ROS, senescence-associated proteins and thrombospondin, an angiogenesis inhibitor. 27 In order to encourage DFUs to heal, the repeated stimulation must be managed and the optimum environment for healing created. This is the objective of standard care as recommended by a number of organisations.…”

Thrombospondin 1 in Metabolic Diseases

“…However, high levels of glucose protect CD47 from degradation and promote its association with SIRPα ( 74 , 75 ). Further, protein and mRNA levels of TSP1 and its receptor CD47 were enhanced in endothelial cells isolated from wound lesions of diabetic Wistar rats ( 76 ). By using a CD47-blocking siRNA, endothelial cells show increased proliferation, migration, and tube formation.…”