The Beneficial Pleiotropic Role of Tumour Necrosis Factor Related Apoptosis Inducing Ligand (TRAIL) in Cardiovascular Disease

Background: Cardiovascular disease (CVD) remains the leading cause of death in patients with Type 2 diabetes mellitus (T2DM). Tumour necrosis factor related apoptosis inducing ligand (TRAIL) is a type II transmembrane protein that belongs to the tumour necrosis factor (TNF) superfamily. A growing body of clinical and animal studies highlight TRAILs vasoprotective potential, with TRAIL being linked to myocardial infarction and accelerated vascular calcification, whereas exogenous TRAIL administration exhibits anti-atherosclerotic activity. Throughout these studies, mechanistic information on the precise nature of TRAIL-mediated protection and the identity of the molecular/cellular targets of TRAIL is still extremely limited. The aims of this thesis were to characterise the effects of TRAIL on endothelial cells, as endothelial dysfunction is an important aspect of the atherosclerotic disease process, and to assess the suitability of TRAIL as marker of cardiovascular risk in patients with T2DM. Methods: We examined the effect of TRAIL on various end-points in human aortic endothelial cells (HAECs) in basal and injurious conditions. In tandem with our in vitro investigations, we compared serum TRAIL levels in non-diabetic controls, patients with T2DM and patients with T2DM and CVD. Results: TRAIL reduced oxidative stress in HAECs exposed to the pro-atherogenic conditions induced by oscillatory shear stress (OSS) and pro-inflammatory cytokines. Serum TRAIL was significantly lower in patients with T2DM and CVD compared to patients with T2DM alone or non-diabetic controls. TRAIL predicted the presence of CVD in patients with T2DM with adequate sensitivity and specificity. Conclusions: TRAILs vasoprotective effects in the vasculature are at least partly mediated by its ability to reduce oxidative stress. Pending further investigations in larger trials, serum TRAIL seems to be a useful diagnostic biomarker of clinically relevant CVD in patients with T2DM.</p

Current provision and HCP experiences of remote care delivery and diabetes technology training for people with type 1 diabetes in the UK during the Covid‐19 pandemic

BackgroundThe COVID-19 pandemic has led to the rapid implementation of remote care delivery in type 1 diabetes. We studied current modes of care delivery, healthcare professional experiences and impact on insulin pump training in type 1 diabetes care in the United Kingdom (UK).MethodsThe UK Diabetes Technology Network designed a 48-question survey aimed at healthcare professionals providing care in type 1 diabetes.ResultsOne hundred and forty-three healthcare professionals (48% diabetes physicians, 52% diabetes educators and 88% working in adult services) from approximately 75 UK centres (52% university hospitals, 46% general and community hospitals), responded to the survey. Telephone consultations were the main modality of care delivery. There was a higher reported time taken for video consultations versus telephone (p ConclusionThis survey highlights UK healthcare professional experiences of remote care delivery. While supportive of virtual care models, a number of factors highlighted, especially patient digital literacy, need to be addressed to improve virtual care delivery and device training.</div

TRAIL attenuates RANKL-mediated osteoblastic signalling in vascular cell mono-culture and co-culture models

<div><p>Background and objectives</p><p>Vascular calcification (VC) is a major risk factor for elevated cardiovascular morbidity/mortality. Underlying this process is osteoblastic signalling within the vessel wall involving complex and interlinked roles for receptor-activator of nuclear factor-κB ligand (RANKL), osteoprotegerin (OPG), and tumour necrosis factor-related apoptosis-inducing ligand (TRAIL). RANKL promotes vascular cell osteoblastic differentiation, whilst OPG acts as a neutralizing decoy receptor for RANKL (and TRAIL). With respect to TRAIL, much recent evidence points to a vasoprotective role for this ligand, albeit via unknown mechanisms. In order to shed more light on TRAILs vasoprotective role therefore, we employed <i>in vitro</i> cell models to test the hypothesis that TRAIL can counteract the RANKL-mediated signalling that occurs between the vascular cells that comprise the vessel wall.</p><p>Methods and results</p><p>Human aortic endothelial and smooth muscle cell mono-cultures (HAECs, HASMCs) were treated with RANKL (0–25 ng/mL ± 5 ng/mL TRAIL) for 72 hr. Furthermore, to better recapitulate the paracrine signalling that exists between endothelial and smooth muscle cells within the vessel wall, non-contact transwell HAEC:HASMC co-cultures were also employed and involved RANKL treatment of HAECs (±TRAIL), subsequently followed by analysis of pro-calcific markers in the underlying subluminal HASMCs. RANKL elicited robust osteoblastic signalling across both mono- and co-culture models (e.g. increased BMP-2, alkaline phosphatase/ALP, Runx2, and Sox9, in conjunction with decreased OPG). Importantly, several RANKL actions (e.g. increased BMP-2 release from mono-cultured HAECs or increased ALP/Sox9 levels in co-cultured HASMCs) could be strongly blocked by co-incubation with TRAIL. In summary, this paper clearly demonstrates that RANKL can elicit pro-osteoblastic signalling in HAECs and HASMCs both directly and across paracrine signalling axes. Moreover, within these contexts we present clear evidence that TRAIL can block several key signalling actions of RANKL in vascular cells, providing further evidence of its vasoprotective potential.</p></div

Direct effects of RANKL±TRAIL on OPG levels in HAECs.

<p>HAECs were treated for 72 hr with RANKL (0–25 ng/mL) in the absence and presence of TRAIL (5 ng/mL) and then investigated by qPCR for (<b>A</b>) OPG mRNA. Cells and conditioned media were also harvested for ELISA analysis of (<b>B</b>) OPG cellular protein and (<b>C</b>) released OPG, respectively. *<i>P</i>≤0.05 versus 0 ng/mL RANKL (or control); ^δ<i>P</i>≤0.05 versus 25 ng/mL RANKL.</p

Schematic of the HAEC:HASMC transwell co-culture model.

<p>HAECs within the luminal compartment were treated for 72 hr with RANKL (0–25 ng/mL) in the absence and presence of TRAIL (5 ng/mL). Within the subluminal compartment, HASMCs were then analyzed for key targets (OPG, ALP, Runx2, Sox9, BMP-2.</p

Direct effects of RANKL±TRAIL on BMP-2 levels in HAECs.

<p>HAECs were treated for 72 hr with RANKL (0–25 ng/mL) in the absence and presence of TRAIL (5 ng/mL) and then analyzed by qPCR for (<b>A</b>) BMP-2 mRNA. Cells and conditioned media were also harvested for ELISA analysis of (<b>B</b>) BMP-2 cellular protein and (<b>C</b>) released BMP-2, respectively. *<i>P</i>≤0.05 versus 0 ng/mL RANKL; ^δ<i>P</i>≤0.05 versus corresponding 5 and 25 ng/mL RANKL treatments.</p

Paracrine effects of RANKL±TRAIL on HASMC ALP levels within a HAEC:HASMC co-culture model.

<p>HAECs within the luminal compartment were treated for 72 hr with RANKL (0–25 ng/mL) in the absence and presence of TRAIL (5 ng/mL). Within the subluminal compartment, HASMCs were then harvested and analyzed for (<b>A</b>) ALP mRNA and (<b>B</b>) ALP enzymatic activity by qPCR and ELISA, respectively. *<i>P</i>≤0.05 versus 0 ng/mL RANKL (or control); ^δ<i>P</i>≤0.05 versus 25 ng/mL RANKL.</p

Effect of RANKL±TRAIL (72 hr) upon key osteoblastic targets within HAEC and HASMC mono-cultures, as well as HAEC:HASMC co-cultures.

<p>Effect of RANKL±TRAIL (72 hr) upon key osteoblastic targets within HAEC and HASMC mono-cultures, as well as HAEC:HASMC co-cultures.</p

Paracrine effects of RANKL±TRAIL on HASMC osteoblastic transcription factors within a HAEC:HASMC co-culture model.

<p>HAECs within the subluminal compartment were treated for 72 hr with RANKL (0–25 ng/mL) in the absence and presence of TRAIL (5 ng/mL). Within the subluminal compartment, HASMCs were then analyzed by qPCR for (<b>A</b>) Runx2 and (<b>B</b>) Sox9 mRNA. *<i>P</i>≤0.05 versus 0 ng/mL RANKL (or control); ^δ<i>P</i>≤0.05 versus 25 ng/mL RANKL.</p