Portal de Periódicos da CAPES

Shifting the Focus of Preclinical, Murine Atherosclerosis Studies From Prevention to Late-Stage Intervention

2017; Lippincott Williams & Wilkins; Volume: 120; Issue: 5 Linguagem: Inglês

10.1161/circresaha.116.310101

1524-4571

Richard A. Baylis, Delphine Gomez, Gary K. Owens,

Nuclear Receptors and Signaling

HomeCirculation ResearchVol. 120, No. 5Shifting the Focus of Preclinical, Murine Atherosclerosis Studies From Prevention to Late-Stage Intervention Free AccessArticle CommentaryPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessArticle CommentaryPDF/EPUBShifting the Focus of Preclinical, Murine Atherosclerosis Studies From Prevention to Late-Stage Intervention Richard A. Baylis, Delphine Gomez and Gary K. Owens Richard A. BaylisRichard A. Baylis From the Robert M. Berne Cardiovascular Research Center (R.A.B., D.G., G.K.O.), Department of Biochemistry and Molecular Genetics (R.A.B.), and Department of Molecular Physiology and Biological Physics (D.G., G.K.O.), University of Virginia, Charlottesville. , Delphine GomezDelphine Gomez From the Robert M. Berne Cardiovascular Research Center (R.A.B., D.G., G.K.O.), Department of Biochemistry and Molecular Genetics (R.A.B.), and Department of Molecular Physiology and Biological Physics (D.G., G.K.O.), University of Virginia, Charlottesville. and Gary K. OwensGary K. Owens From the Robert M. Berne Cardiovascular Research Center (R.A.B., D.G., G.K.O.), Department of Biochemistry and Molecular Genetics (R.A.B.), and Department of Molecular Physiology and Biological Physics (D.G., G.K.O.), University of Virginia, Charlottesville. Originally published3 Mar 2017https://doi.org/10.1161/CIRCRESAHA.116.310101Circulation Research. 2017;120:775–777Could improving the design of preclinical murine atherosclerosis studies help increase the success rate of cardiovascular clinical trials? In this Viewpoint, the authors advocate for a change from prevention to intervention study designs and rigorous lesion analyses that they argue will enhance the translational potential of murine atherosclerosis studies.Clinical management of patients with coronary artery disease (CAD) has made great progress by reducing risk factors like low-density lipoprotein cholesterol level, hypertension, and diabetes mellitus. However, late-stage events associated with coronary atherosclerosis including myocardial infarction still account for ≈16% of worldwide mortality and are forecasted to increase in prevalence.1 These events arise from 3 main processes: plaque rupture, plaque erosion, and shedding of calcific nodules. Plaque rupture accounts for the majority of CAD and typically occurs in vulnerable atherosclerotic lesions termed thin-capped fibroatheromas,2 characterized by thin fibrous caps ( 80% of SMC-derived cells lack detectable Acta2 expression and (2) >30% of Lgals3+ cells are SMC derived in both mouse and human lesions.9 By combining lineage tracing with simultaneous SMC-specific gene knockout of pluripotency factors Klf4 or Oct4, we have shown that SMC can play a critical role in the pathogenesis of late-stage lesions, which can be either atheroprotective or atheropromoting depending on the nature of their phenotypic transitions. For example, Klf4-dependent transitions, including formation of SMC-derived macrophage-marker+ foam cells,9 exacerbated lesion pathogenesis, whereas Oct4-dependent transitions were atheroprotective, being required for migration and stable investment of SMC into the fibrous cap.10 Taken together, these results highlight the critical importance of identifying factors and mechanisms that promote plaque stabilizing atheroprotective changes within SMC and other major cell types within lesions and therapeutic approaches that can induce such changes. The findings also highlight the importance of lineage tracing in atherosclerosis. Because of significant plasticity in multiple lesion cell types, there is substantial ambiguity in cell identification when using traditional markers (eg, Acta2, CD68, and CD31), which can only be overcome by the use of rigorous lineage-tracing techniques.In addition to studying more relevant pathological parameters, scientists should consider implementing more relevant study designs. Currently, the overwhelming majority of atherosclerosis studies implement models of prevention. Namely, the researcher deletes gene x or provides drug y before—or in the early stages of—lesion formation, and therefore, the therapeutic agent or genetic manipulation is given to young, healthy mice and is present throughout lesion development (Figure). With these prevention models, we have learned a great deal about the steps of atherosclerosis development—determining the key cellular players and identifying thousands of genes that can alter lesion accumulation—but unfortunately, this has translated to few novel therapies. Therefore, although prevention models may be suitable for assessing therapeutic feasibility, we advocate for additional interventional studies before initiation of clinical trials. Here, mice are treated with the therapy or gene knockout only after they have established advanced atherosclerosis. By providing the therapy at this stage and then assessing for changes in the indices of lesion stability as described above, we obtain a better prediction of how the intervention will impact the processes regulating late-stage lesion vulnerability and, hopefully, gain better insight into its impact on advanced human lesions. Clearly, different processes are at work during lesion development versus late-stage disease. For example, elegant studies from Filip Swirski's group12 have shown that monocyte recruitment drives early lesion macrophage accumulation but that as the lesion matures the accumulation becomes a function of local macrophage proliferation. These fundamental biological differences between disease stages may indeed provide one possible explanation for the poor translation of prevention studies to patients with advanced disease. It should be noted that idea of implementing interventional models to the study of atherosclerosis is not novel. Indeed, there are many excellent examples in the literature using both pharmacological (eg, delivering collagen IV-targeted nanoparticles containing proresolving peptide, Ac2-26)13 and genetic (eg, SMC-specific knockout of Akt1 after 16 weeks of Western diet)14 interventions validating its importance to the field—but we feel that it remains heavily underutilized. Another study design that has been used to study the processes critical in late-stage atherosclerosis is the regression model. These are variations of the intervention model in which atherosclerosis is induced by chronic hypercholesterolemia but then regressed by either normalizing specific lipid components or transplanting diseased vessels into healthy organisms. These models have been helpful to identify the processes that may be reversible in advanced atherosclerosis like CCR7-mediated macrophage egress15 or reverse cholesterol transport and may prove valuable when investigating strategies to encourage these processes.Download figureDownload PowerPointFigure. Proposed modifications to preclinical pipeline for experimental atherosclerosis studies. Prevention studies (A) may represent a good proof-of-principle for a novel therapeutic or gene knockout but in the setting of atherosclerosis may not translate to older patients with established disease. Instead, we should implement intervention studies (B), which we argue will better predict the effect of a therapeutic strategy for treating humans with advanced atherosclerotic lesions. Both of these approaches should analyze parameters that not only provide information about lesion size but also investigate multiple key cellular processes implicated in lesion vulnerability in humans. In addition, it is critical to identify innovative ways for validating results in preclinical animal studies to the extent possible including use of classic immunohistological analyses of human autopsy specimens and large-scale genomic approaches (C). Of course, the final validation of new cardiovascular disease therapies will require well-designed clinical trials (D).In summary, we hope to motivate investigators to re-evaluate the way we apply mouse models to the study of atherosclerosis. With the residual risk for atherosclerotic complications that remains despite current standard of care, there exists a critical need for therapies that not only focus on limiting destabilization but also seek to promote better inflammatory resolution, healing, and overall plaque stabilization. We think that the first step toward achieving this goal is to begin studying potential therapeutics using interventional models in mice and to rigorously characterize their effect on processes critical in maintaining lesion stability. Although no animal model completely recapitulates human disease, we can and need to do a better job of matching our experimental animal model designs to the unmet clinical needs.Sources of FundingThis work was supported by R01 grants HL057353, HL098538, and HL087867 to G.K. Owens and the American Heart Association 15SDG25860021 for D. Gomez.DisclosuresNone.FootnotesThe opinions expressed in this article are not necessarily those of the editors or of the American Heart Association.Correspondence to Gary K. Owens, PhD, University of Virginia School of Medicine, 415 Lane Rd, PO Box 801394, Room 1332, Medical Research Bldg 5, Charlottesville, VA 22908. E-mail [email protected]References1. GBD 2015 DALYs and HALE Collaborators. Global, regional, and national disability-adjusted life-years (DALYs) for 315 diseases and injuries and healthy life expectancy (HALE), 1990–2015: a systematic analysis for the Global Burden of Disease Study 2015.Lancet. 2016; 388:1603–1658.CrossrefMedlineGoogle Scholar2. Kolodgie FD, Virmani R, Burke AP, Farb A, Weber DK, Kutys R, Finn AV, Gold HK. Pathologic assessment of the vulnerable human coronary plaque.Heart. 2004; 90:1385–1391. doi: 10.1136/hrt.2004.041798.CrossrefMedlineGoogle Scholar3. Hay M, Thomas DW, Craighead JL, Economides C, Rosenthal J. Clinical development success rates for investigational drugs.Nat Biotechnol. 2014; 32:40–51. doi: 10.1038/nbt.2786.CrossrefMedlineGoogle Scholar4. Johnson JL, Jackson CL. Atherosclerotic plaque rupture in the apolipoprotein E knockout mouse.Atherosclerosis. 2001; 154:399–406.CrossrefMedlineGoogle Scholar5. Calara F, Silvestre M, Casanada F, Yuan N, Napoli C, Palinski W. Spontaneous plaque rupture and secondary thrombosis in apolipoprotein E-deficient and LDL receptor-deficient mice.J Pathol. 2001; 195:257–263. doi: 10.1002/path.915.CrossrefMedlineGoogle Scholar6. Bentzon JF, Sondergaard CS, Kassem M, Falk E. Smooth muscle cells healing atherosclerotic plaque disruptions are of local, not blood, origin in apolipoprotein E knockout mice.Circulation. 2007; 116:2053–2061. doi: 10.1161/CIRCULATIONAHA.107.722355.LinkGoogle Scholar7. Sasaki T, Kuzuya M, Nakamura K, Cheng XW, Shibata T, Sato K, Iguchi A. A simple method of plaque rupture induction in apolipoprotein E-deficient mice.Arterioscler Thromb Vasc Biol. 2006; 26:1304–1309. doi: 10.1161/01.ATV.0000219687.71607.f7.LinkGoogle Scholar8. Thusen von der JH. Induction of atherosclerotic plaque rupture in apolipoprotein E-/- mice after adenovirus- mediated transfer of p53.Circulation. 2002; 105:2064–2070.LinkGoogle Scholar9. Shankman LS, Gomez D, Cherepanova OA, Salmon M, Alencar GF, Haskins RM, Swiatlowska P, Newman AA, Greene ES, Straub AC, Isakson B, Randolph GJ, Owens GK. KLF4-dependent phenotypic modulation of smooth muscle cells has a key role in atherosclerotic plaque pathogenesis.Nat Med. 2015; 21:628–637. doi: 10.1038/nm.3866.CrossrefMedlineGoogle Scholar10. Cherepanova OA, Gomez D, Shankman LS, et al. Activation of the pluripotency factor OCT4 in smooth muscle cells is atheroprotective.Nat Med. 2016; 22:657–665. doi: 10.1038/nm.4109.CrossrefMedlineGoogle Scholar11. Feil S, Fehrenbacher B, Lukowski R, Essmann F, Schulze-Osthoff K, Schaller M, Feil R. Transdifferentiation of vascular smooth muscle cells to macrophage-like cells during atherogenesis.Circ Res. 2014; 115:662–667. doi: 10.1161/CIRCRESAHA.115.304634.LinkGoogle Scholar12. Robbins CS, Hilgendorf I, Weber GF, et al. Local proliferation dominates lesional macrophage accumulation in atherosclerosis.Nat Med. 2013; 19:1166–1172. doi: 10.1038/nm.3258.CrossrefMedlineGoogle Scholar13. Fredman G, Kamaly N, Spolitu S, Milton J, Ghorpade D, Chiasson R, Kuriakose G, Perretti M, Farokhzad O, Farokzhad O, Tabas I. Targeted nanoparticles containing the proresolving peptide Ac2-26 protect against advanced atherosclerosis in hypercholesterolemic mice.Sci Transl Med. 2015; 7:275ra20. doi: 10.1126/scitranslmed.aaa1065.CrossrefMedlineGoogle Scholar14. Rotllan N, Wanschel AC, Fernández-Hernando A, Salerno AG, Offermanns S, Sessa WC, Fernández-Hernando C. Genetic evidence supports a major role for Akt1 in VSMCs during atherogenesis.Circ Res. 2015; 116:1744–1752. doi: 10.1161/CIRCRESAHA.116.305895.LinkGoogle Scholar15. Feig JE, Shang Y, Rotllan N, Vengrenyuk Y, Wu C, Shamir R, Torra IP, Fernandez-Hernando C, Fisher EA, Garabedian MJ. Statins promote the regression of atherosclerosis via activation of the CCR7-dependent emigration pathway in macrophages.PLoS One. 2011; 6:e28534. doi: 10.1371/journal.pone.0028534.CrossrefMedlineGoogle Scholar Previous Back to top Next FiguresReferencesRelatedDetailsCited By Sakic A, Chaabane C, Ambartsumian N, Klingelhöfer J, Lemeille S, Kwak B, Grigorian M and Bochaton-Piallat M (2020) Neutralization of S100A4 induces stabilization of atherosclerotic plaques: role of smooth muscle cells, Cardiovascular Research, 10.1093/cvr/cvaa311, 118:1, (141-155), Online publication date: 7-Jan-2022. Madonna R, Doria V, Görbe A, Cocco N, Ferdinandy P, Geng Y, Pierdomenico S and De Caterina R (2020) Co‐expression of glycosylated aquaporin‐1 and transcription factor NFAT5 contributes to aortic stiffness in diabetic and atherosclerosis‐prone mice, Journal of Cellular and Molecular Medicine, 10.1111/jcmm.14843, 24:5, (2857-2865), Online publication date: 1-Mar-2020. Frégeau G, Sarduy R, Elimam H, Esposito C, Mellal K, Ménard L, Leitão da Graça S, Proulx C, Zhang J, Febbraio M, Soto Y, Lubell W, Ong H and Marleau S (2020) Atheroprotective and atheroregressive potential of azapeptide derivatives of GHRP-6 as selective CD36 ligands in apolipoprotein E-deficient mice, Atherosclerosis, 10.1016/j.atherosclerosis.2020.06.010, 307, (52-62), Online publication date: 1-Aug-2020. Pustlauk W, Westhoff T, Claeys L, Roch T, Geißler S and Babel N (2020) Induced osteogenic differentiation of human smooth muscle cells as a model of vascular calcification, Scientific Reports, 10.1038/s41598-020-62568-w, 10:1, Online publication date: 1-Dec-2020. Harman J and Jørgensen H (2019) The role of smooth muscle cells in plaque stability: Therapeutic targeting potential, British Journal of Pharmacology, 10.1111/bph.14779, 176:19, (3741-3753), Online publication date: 1-Oct-2019. Zhou Y, Liu X, Huang N and Chen Y (2018) Magnesium ion leachables induce a conversion of contractile vascular smooth muscle cells to an inflammatory phenotype, Journal of Biomedical Materials Research Part B: Applied Biomaterials, 10.1002/jbm.b.34192, 107:4, (988-1001), Online publication date: 1-May-2019. Gomez D, Baylis R, Durgin B, Newman A, Alencar G, Mahan S, St. Hilaire C, Müller W, Waisman A, Francis S, Pinteaux E, Randolph G, Gram H and Owens G (2018) Interleukin-1β has atheroprotective effects in advanced atherosclerotic lesions of mice, Nature Medicine, 10.1038/s41591-018-0124-5, 24:9, (1418-1429), Online publication date: 1-Sep-2018. Allahverdian S, Chaabane C, Boukais K, Francis G and Bochaton-Piallat M (2018) Smooth muscle cell fate and plasticity in atherosclerosis, Cardiovascular Research, 10.1093/cvr/cvy022, 114:4, (540-550), Online publication date: 15-Mar-2018. Bachmann J, Baumgart S, Uryga A, Bosteen M, Borghetti G, Nyberg M and Herum K (2022) Fibrotic Signaling in Cardiac Fibroblasts and Vascular Smooth Muscle Cells: The Dual Roles of Fibrosis in HFpEF and CAD, Cells, 10.3390/cells11101657, 11:10, (1657) March 3, 2017Vol 120, Issue 5 Advertisement Article InformationMetrics © 2017 American Heart Association, Inc.https://doi.org/10.1161/CIRCRESAHA.116.310101PMID: 28254801 Originally publishedMarch 3, 2017 Keywordsatherosclerosisdiabetes mellitussmooth muscletranslational medical researchcollagencoronary diseasemyocardial infarctionPDF download Advertisement SubjectsBasic Science ResearchMechanismsTranslational StudiesVascular Biology