Arterial Delivery of VEGF-C Stabilizes Atherosclerotic Lesions

HomeCirculation ResearchVol. 128, No. 2Arterial Delivery of VEGF-C Stabilizes Atherosclerotic Lesions Free AccessLetterPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyRedditDiggEmail Jump toFree AccessLetterPDF/EPUBArterial Carlos Silvestre-Roig, Patricia Lemnitzer, Julie Gall, Simon Schwager, Albulena Toska, Laurent Yvan-Charvet, Michael Detmar and Oliver Soehnlein Silvestre-RoigCarlos Silvestre-Roig Institute for Cardiovascular Prevention (IPEK), Klinikum der LMU München (C.S.-R., P.L., O.S.). German Center Research (DZHK), Munich Search more papers by this author , LemnitzerPatricia Lemnitzer GallJulie Gall (INSERM) U1065, Université Côte d’Azur, Centre Méditerranéen de Médecine Moléculaire (C3M), Atip-Avenir, Fédération Hospitalo-Universitaire (FHU) Oncoage, Nice (J.G., L.Y.-C.). SchwagerSimon Schwager Pharmaceutical Sciences, Swiss Federal Technology, ETH Zurich (S.S., M.D.). ToskaAlbulena Toska https://orcid.org/0000-0002-4519-9493 Medical Microbiology, Immunology Hygiene, Technische Universität (TUM) (A.T.). Yvan-CharvetLaurent Yvan-Charvet DetmarMichael SoehnleinOliver Correspondence to: Soehnlein, MD, PhD, Prevention, Ludwig-Maximilians-University Munich, Pettenkoferstr. 9, 80336 Munich. Email E-mail Address: [email protected] https://orcid.org/0000-0002-7854-0694 Physiology Pharmacology (FyFa), Karolinska Institutet, Stockholm (O.S.). Originally published19 Nov 2020https://doi.org/10.1161/CIRCRESAHA.120.317186Circulation Research. 2021;128:284–286Other version(s) articleYou are viewing the most recent version article. Previous versions: November 19, 2020: Ahead Print Cell stress elicited intracellular lipid accumulation causes cell death, a major driver atherosclerosis. Expansion necrotic core fibrous cap (FC) thinning as consequence smooth muscle (SMC) death hallmarks vulnerable plaques. Therefore, therapeutic strategies limit SMC rarefication key prevent atheroprogression. While clinical trials support overall viability therapeutically targeting arterial inflammation, they also epitomize shortcomings such strategies, systemic immunomodulation exhibits adverse side effects including impaired host defense.1 Hence, it is interest preventive cardiology identify means boycott inflammation locally.Arterial lymphatics represent exit routes lesional leukocytes serve channels reverse cholesterol transport, thus limiting atheroprogression.2,3 Here, we hypothesized that targeted delivery VEGF (vascular endothelial growth factor)-C, dominant factor lymphatics, preexisting lesions reverses accumulation, reduces intimal confers plaque stability. The F8 antibody specifically targets extra domain A fibronectin, an isoform accumulates in inflamed tissues atherosclerotic lesions. Extra A’s absence steady-state renders suitable target site-specifically deliver cytokines hence tested ability F8-VEGF-C conjugates stabilize lesions.Advanced carotid artery Apoe−/− mice were generated previously described.4 All experiments approved local ethics committee performed accordance with institutional guidelines. Only female included study males do not develop advanced after cast insertion. Analyses made blindly. Images representing mean value certain readout chosen representative images.The F8-antibody showed preferential binding tissue hypercholesterolemic compared other organs (Figure [A], left). Similarly, bound FC area human right). To investigate on atherosclerosis, treated [B], This treatment did significantly affect plasma levels or circulating leukocyte counts (not shown) while strikingly increased middle/right). In contrast receiving vehicle only, lesion sizes increase baseline [C]). addition, improved signs instability exemplified reduced [C]), thicker FCs [C]) accompanied expansion collagen content within shown). Contrasting our initial hypothesis, however, alter lymph vessel area, macrophage burden, egress Instead, SMCs analysis cells significant decrease loading but macrophages F8-VEGF-C–treated mice. Overall, these data led us speculate lowers SMCs, thereby improving their survival.Download figureDownload PowerPointFigure. Arterial improves stability.A–C, Advanced insertion shear modifier around A, Representative images indicated (left) (right). B, Schematic representation arterial-directed factor)-C (left). micrographs showing (SMCs) VEGF-C. Displayed quantification VEGF-C+ vehicle-treated VEGF-C–treated C, hematoxylin-eosin–stained mouse intima media ratio, thickness, lipid-laden SMCs. Confocal immunofluorescence lipids. White arrowheads point at droplets SMA+ area. D, Cholesterol efflux [3H]-cholesterol-loaded was medium containing 5% polyethylene glycol (PEG)-HDL (high-density lipoprotein). stimulated rhVEGF-C (recombinant VEGF) (500 ng/mL, 24 h) represented relative LXR (liver X receptor) agonist (TO901317)–treated Quantification ratio Abca1 Gapdh mRNA quantified Digital Drop polymerase chain reaction Both parts show 2 independent each. E, pretreated (24 incubated 7-ketocholesterol (200 μg/mL) amounts another h. measured using propidium iodide (PI) uptake western blot probed antibodies β-actin CHOP (middle). SMA+TUNEL+ (terminal deoxynucleotidyl transferase dUTP nick end labeling) over total Data mean±SEM. analyzed GraphPad Prism. Normality assessed D’Agostino Pearson test. Significance Mann-Whitney test (B) (D, right); Kruskal-Wallis Dunn correction thickness (C) left) (E, left); 1-way ANOVA Tukey remaining (C); unpaired 2-sided t F, Scheme summarizing proposed mechanism. G, Table used study. ABCA1 indicates ATP-binding cassette transporter A1; α-SMA, actin; CHOP, CCAAT-enhancer-binding protein homologous protein; DAPI, 4′,6-diamidino-2-phenylindole; ER, endoplasmic reticulum; FITC, fluorescein isothiocyanate; HFD, high fat diet; L, lumen.To idea, engaged ex vivo studies. Although oxidized LDL (low-density lipoprotein) affected shown), favored similar extent positive control [D], left), observation can least part be explained expression (ATP-binding A1) response Lipid toxicity associates through reticulum promoting destabilization. evaluate reduce lipid-mediated exposed presence Interestingly, abolished evoked [E], known mediate its actions VEGFR (VEGF 3, receptor could Neuropilin 1 neuropilin have been suggested some indeed receptors subsets recently published scRNAseq (single RNA sequencing) database SMCs5 Using siRNA (small interfering RNA)-mediated knockdown inhibited VEGF-C-induced Of note, family member VEGF-A unable induce induced Moreover, protective effect range protection governed inhibitor salubrinal associated transcription C/EBP (CCAAT-enhancer-binding proteins) Homologous Protein middle), important trigger stress. vivo, right).Taken together, here report beneficial stability rescuing atheroprotective unexpectedly diminished stress, ultimately prevented [F]). use antibody-based like one employed may promising effects.Nonstandard Abbreviation AcronymsFCfibrous capSMCsmooth cellVEGFvascular factorSources FundingDeutsche Forschungsgemeinschaft (SO876/11-1, SFB914 TP B8, SFB1123 A6, B5), Vetenskapsrådet (2017-01762), Else-Kröner-Fresenius Stiftung, Leducq foundation.Disclosures findings available from corresponding upon reasonable request.FootnotesFor Sources Funding Disclosures, see page 285.Correspondence oliver.[email protected]comReferences1. 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Andone S, Bajko Z, Motataianu Mosora Balasa R Role Biomarkers Atherothrombotic Stroke—A Systematic Review, International Journal Molecular 10.3390/ijms22169032, 22:16, (9032) Miyazaki Hypercholesterolemia Defects: Chicken Egg?, Frontiers Medicine, 10.3389/fcvm.2021.701229, 8 Dabravolski Khotina V, Omelchenko Kalmykov V Orekhov Family Atherosclerosis Development Its Potential Treatment Targets, 10.3390/ijms23020931, 23:2, (931) January 22, 2021Vol Issue 2Article InformationMetrics © 2020 American Association, Inc.https://doi.org/10.1161/CIRCRESAHA.120.317186PMID: 33210556 publishedNovember KeywordscytokineleukocyteinflammationatherosclerosisfibronectinPDF download Advertisement SubjectsAnimal Models Human DiseaseAtherosclerosisBasic Science ResearchInflammationVascular Biology