bioRxiv Science⌕ Search

Biology subjects

Solly, E.

Publications and source records attributed to Solly, E..

2 recordsLinked to original sources

Development of a novel murine model of in-stent neoatherosclerosis

ObjectiveIn-stent neoatherosclerosis is a phenomenon of percutaneous coronary intervention with stenting. Whilst similar to de novo atherosclerosis, it develops rapidly over 1-5 years rather than over a lifetime. No preclinical small animal models exist that allow full elucidation of neoatherosclerosis biology and future treatments. The aim of this study was to establish and validate a novel murine model of in-stent neoatherosclerosis. Approach and ResultsMurine stainless-steel stents (2.5 x 0.7 mm) were deployed into donor descending aortas of atherosclerosis-prone apolipoprotein (Apo)e-/- mice, then carotid-interposition grafted into Apoe-/- recipients. Mice (n=6-8/group) received chow or a high cholesterol diet (HCD) for 7- or 28-days post-surgery. Multimodal intravascular imaging, simultaneously combining optical coherence tomography (OCT, plaque burden) and fluorescence for indocyanine green (ICG, plaque instability), visualized in-stent neoatherosclerosis across the entire length of the stented site. Histological analyses revealed that stented vessels from mice fed HCD had neointimas with prominent lipid cores and an elevated CD68+ macrophage content, similar to human neoatherosclerosis. Mice fed chow post-stenting had distinctly different neointimas that were smooth muscle cell rich, resembling neointimal hyperplasia. Consistent with this, flow cytometry revealed a higher content of monocytes/macrophages and dendritic cells in stented aortas from mice fed HCD than in non-stented aortas. ConclusionWe have developed and validated the first murine model that replicates the unique characteristics of human in-stent neoatherosclerosis. This project has implications for exploring the mechanisms that promote neoatherosclerosis and testing targeted new therapies. RESEARCH PERSPECTIVEO_ST_ABSWhat Is New?C_ST_ABSO_LIWe have developed and validated a novel murine model of in-stent neoatherosclerosis, presenting a new platform that will facilitate the discovery of novel mechanistic targets of in-stent neoatherosclerosis and preventative therapies. C_LIO_LIThis model develops lesions with a similar morphology to human in-stent neoatherosclerosis and distinct to in-stent neointimal hyperplasia, with higher extracellular lipid and macrophage content and proportionately less smooth muscle cells. C_LIO_LIWe show a first-time visualization of murine in-stent neoatherosclerosis using bimodal intravascular imaging with simultaneous capture of structural information (optical coherence tomography, plaque burden) and the distribution of areas of plaque instability (high-sensitivity fluorescence, indocyanine green) within the plaque. C_LI What new question does this study raise?O_LIHow can the utility of this novel model be maximized as a platform for discovering novel agents that prevent in-stent neoatherosclerosis? C_LI What question should be addressed next?O_LIAre there unique mechanisms of in-stent neoatherosclerosis, distinct to de novo atherosclerosis, that can be specifically targeted to prevent disease and ultimately increase stent performance? C_LI

cell biology↗

Reconstituted high-density lipoproteins rescue diabetes-impaired endothelial cell metabolic reprograming and angiogenic responses to hypoxia

ObjectiveImpaired angiogenic responses to ischemia underlie diabetic vascular complications. Reconstituted high-density lipoproteins (rHDL) have proangiogenic effects in diabetes. The pyruvate dehydrogenase kinase 4 (PDK4)/pyruvate dehydrogenase complex (PDC) axis is an oxygen-conserving mechanism that preserves EC functions in hypoxia. We aimed to determine the role of the PDK4/PDC axis in angiogenesis, the effect of diabetes on its regulation in response to ischemia, and in the proangiogenic properties of rHDL. Approach and ResultsIn a murine wound healing model, PDK4 and pPDC were elevated early (24h) post induction of wound ischemia in non-diabetic wounds, which did not occur in diabetic mice. Topical rHDL rescued this impairment, enhancing PDK4 (68%, P<0.05) and pPDC (165%, P<0.01) in diabetic wounds. In parallel, wound neovascularization (62%, P<0.05) and closure (154%, P<0.0001) were increased in diabetic rHDL-treated wounds. In vitro, PDK4 and pPDC levels were increased in ECs exposed to hypoxia (65%, 70% respectively, P<0.05). High glucose did not elicit a further step-wise induction in PDK4/pPDC, with aberrant increases in mitochondrial respiration (19%, P<0.05), coupled with impaired EC angiogenic functions. Importantly, rHDL increased PDK4 and pPDC two-fold, returning mitochondrial respiration and EC angiogenic functions to normal glucose levels. In vitro, PDK4 siRNA knockdown attenuated the proangiogenic effects of rHDL. In vivo PDK4 inhibition ameliorated topical rHDL-mediated increases in wound angiogenesis and closure. Using chromatin immunoprecipitation, rHDL increased forkhead box O1 (FOXO1) binding to the PDK4 promoter and suppressed FOXO1 phosphorylation, presenting FOXO1 as a mechanism for the induction of PDK4 by rHDL. ConclusionThe PDK4/PDC axis response to ischemia is impaired in diabetes and important for the proangiogenic effects of rHDL.

cell biology↗