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Nankivell, V.

Publications and source records attributed to Nankivell, V..

3 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↗

Multifunctional biomimetic porphyrin-lipid nanoparticles - novel nanoscale theranostics for atherosclerotic cardiovascular disease

BackgroundHigh-density lipoprotein (HDL) nanoagents have unrealized potential for atherosclerosis theranostics. Porphyrin-lipid HDL mimetic nanoparticles (Por-HDL-NPs) incorporate porphyrin-lipid which permits near infrared fluorescence imaging and positron emission tomography (PET) through chelation of Copper-64 (64Cu). The outer shell contains apolipoprotein A-I mimetic peptide R4F that interacts with scavenger receptor SR-BI, enabling macrophage targeting and therapeutic effects. We leveraged the theranostic properties of Por-HDL-NPs for testing in atherosclerosis. Methods and ResultsIn vitro, Por-HDL-NPs were internalised by immortalised bone marrow-derived macrophages (iBMDMs), visualised via fluorescence microscopy and flow cytometry. Por-HDL-NPs increased cholesterol efflux from [3H]-cholesterol-loaded iBMDMs, (49%, P<0.05), compared to reconstituted HDL. Incubation of iBMDMs with Por-HDL-NPs reduced mRNA levels of inflammatory mediators Il-1{beta} (88%), Il-18 (54%) and Ccl5 (75%), and protein secretion of IL-1{beta} (69%) and CCL5 (82%), P<0.05. Por-HDL-NPs suppressed inflammasome components Nlrp3 (69%) and Asc (36%), P<0.05. Studies using siRNA deletion of SR-B1 and methyl-{beta}-cyclodextrin, revealed the anti-inflammatory properties of Por-HDL-NPs were independent of SR-B1 and cholesterol efflux. However, Por-HDL-NPs suppressed activation of inflammatory transcription factor NF-{kappa}B (53%, P<0.05). In Apoe-/- mice, PET imaging showed 64Cu-Por-HDL-NPs localised in hearts and detected increases in plaque over time with high-cholesterol diet. Por-HDL-NP fluorescence was visualised in aortic sinus plaques, co-localised with CD68+ macrophages, and by fluorescence IVIS imaging in aortic arch plaque. Por-HDL-NP-treated mice had smaller early-stage (22%) and unstable plaques (52%) and fewer circulating monocytes (32%) than control PBS-treated mice, P<0.05 for all. ConclusionsPor-HDL-NPs have theranostic properties, exhibiting both multi-modal imaging capabilities for identifying plaque and athero-protective therapeutic effects. Clinical PerspectiveO_ST_ABSWhat is new?C_ST_ABSO_LIPorphyrin high-density lipoprotein (HDL) mimetic nanoparticles (Por-HDL-NPs) have theranostic application in atherosclerosis. C_LIO_LIPor-HDL-NPs are internalized by macrophages in vitro and plaque macrophages in vivo, enabling the visualization of atherosclerosis by both positron emission tomography and multiple fluorescence imaging modalities. C_LIO_LIPor-HDL-NPs exhibit atheroprotective effects and suppress inflammation, promote cholesterol efflux, reduce atherosclerotic plaque development and lower the number of circulating monocytes. C_LI What are the clinical implications?O_LIThe PET imaging and plaque targeting capabilities of Por-HDL-NPs have implications for improved non-invasive tracking of human atherosclerosis development. C_LIO_LIThe excellent fluorescence imaging and plaque targeting properties of Por-HDL-NPs have clinical significance for improved detection of early-stage plaque using intravascular imaging strategies. C_LIO_LIPor-HDL-NPs provide therapeutic capabilities that target plaque directly, independent of lipid-lowering, suggestive of their potential to provide benefit on top of current lipid-lowering strategies. C_LI

cell biology↗