In rodents with acute myocardial infarction (AMI), high mobility group box 1 (HMGB1) injection has produced controversial results. Given the lack of data in large mammals, we searched the dose that would promote angiogenesis and expression of specific regenerative genes in sheep with AMI (protocol 1) and, subsequently, use this dose to study long-term effects on infarct size and left ventricular (LV) function (protocol 2). Protocol 1: Sheep with AMI received 250 μg (high-dose, n = 7), 25 μg (low-dose, n = 7) HMGB1, or PBS (placebo, n = 7) in 10 intramyocardial injections (0.2 ml each) in the peri-infarct area. Seven days later, only the high-HMGB1-dose group exhibited higher microvascular densities, Ki67-positive cardiomyocytes, and overexpression of VEGF, Ckit, Tbx20, Nkx2.5, and Gata4. Protocol 2: Sheep with AMI received HMGB1 250 μg (n = 6) or PBS (n = 6). At 60 days, HMGB1-treated sheep showed smaller infarcts (8.5 ± 2.11 vs. 12.2 ± 1.97% LV area, P < 0.05, ANOVA-Bonferroni) and higher microvascular density (capillaries, 1798 ± 252 vs. 1266 ± 250/mm2; arterioles, 18.3 ± 3.9 vs. 11.7 ± 2.2/mm2; both P < 0.01). Echocardiographic LV ejection fraction, circumferential shortening, and wall thickening increased from day 3 to 60 with HMGB1 (all P < 0.05). Conclusion: in ovine AMI, high-dose HMGB1 induces angio-arteriogenesis, reduces infarct size, and improves LV function at 2 months post-treatment.
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We thank veterinarians María Inés Besansón and Pedro Iguain for anesthetic management and animal house assistants Juan Carlos Mansilla, Osvaldo Sosa, and Juan Ocampo for dedicated care of the animals. We also thank Julio Martínez, Fabián Gauna, and Rosana Valverdi for technical help.
Supported by grant 2012-1729 from the National Agency for the Promotion of Science and Technology (ANPCyT), Ministry of Science, Technology and Innovative Production (MINCyT) of Argentina.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All procedures were carried out in accordance with the Guide for Care and Use of Laboratory Animals, published by the US National Institutes of Health (NIH Publication No. 85–23, revised 1996) and approved and monitored by the Laboratory Animal Care and Use Committee (CICUAL) of the Favaloro University (approval # DCT0157-12). The manuscript does not contain clinical studies or patient data.
Roth GA, Johnson C, Abajobir A, Abd-Allah F, Abera SF, Abyu G, et al. Global, regional, and national burden of cardiovascular diseases for 10 causes, 1990 to 2015. J Am Coll Cardiol. 2017;70:1–25.CrossRefPubMedPubMedCentralGoogle Scholar
Tiyyagura SR, Pinney SP. Left ventricular remodeling after myocardial infarction: past, present, and future. Mt Sinai J Med. 2006;73:840–51.PubMedGoogle Scholar
Stone GW, Selker HP, Thiele H, Patel MR, Udelson JE, Ohman EM, et al. Relationship between infarct size and outcomes following primary PCI: patient-level analysis from 10 randomized trials. J Am Coll Cardiol. 2016;67:1674–83.CrossRefGoogle Scholar
Gnavi R, Rusciani R, Dalmasso M, Giammaria M, Anselmino M, Roggeri DP, et al. Gender, socioeconomic position, revascularization procedures and mortality in patients presenting with STEMI and NSTEMI in the era of primary PCI. Differences or inequities? Int J Cardiol. 2014;176:724–30.CrossRefPubMedGoogle Scholar
De Luca G, Petrelli A, Landriscina T, Gnavi R, Giammaria M, Costa G. Geographic and socioeconomic differences in access to revascularization following acute myocardial infarction. Eur J Pub Health. 2016;26:760–5.CrossRefGoogle Scholar
Rebouças JS, Santos-Magalhães NS, Formiga FR. Cardiac regeneration using growth factors: advances and challenges. Arq Bras Cardiol. 2016;107:271–5.PubMedPubMedCentralGoogle Scholar
Awada HK, Hwang MP, Wang Y. Towards comprehensive cardiac repair and regeneration after myocardial infarction: aspects to consider and proteins to deliver. Biomaterials. 2016;82:94–112.CrossRefGoogle Scholar
Guo M, Shi JH, Wang PL, Shi DZ. Angiogenic growth factors for coronary artery disease: current status and prospects. J Cardiovasc Pharmacol Ther. 2018;23:130–41.CrossRefPubMedGoogle Scholar
Bianchi ME, Agresti A. HMG proteins: dynamic players in gene regulation and differentiation. Curr Opin Genet Dev. 2005;15:496–506.CrossRefPubMedGoogle Scholar
Raucci A, Di Maggio S, Scavello F, D’Ambrosio A, Bianchi M, Capogrossi MC. The Janus face of HMGB1 in heart disease: a necessary update. Cell Mol Life Sci. 2019;76:211–29.CrossRefPubMedGoogle Scholar
Kitahara T, Takeishi Y, Harada M, Niizeki T, Suzuki S, Sasaki T, et al. Highmobility group box 1 restores cardiac function after myocardial infarction in transgenic mice. Cardiovasc Res. 2008;80:40–6.CrossRefPubMedGoogle Scholar
Takahashi K, Fukushima S, Yamahara K, Yashiro K, Shintani Y, Coppen SR, et al. Modulated inflammation by injection of high-mobility group box 1 recovers post-infarction chronically failing heart. Circulation. 2008;118(Suppl 1):S106–14.CrossRefPubMedGoogle Scholar
Zhou X, Hu X, Xie J, Xu C, Xu W, Jiang H. Exogenous high-mobility group box 1 protein injection improves cardiac function after myocardial infarction: involvement of Wnt signaling activation. J Biomed Biotechnol. 2012;2012:1–5. https://doi.org/10.1155/2012/743879.CrossRefGoogle Scholar
Di Maggio S, Milano G, De Marchis F, D’Ambrosio A, Bertolotti M, Palacios BS, et al. Non-oxidizable HMGB1 induces cardiac fibroblasts migration via CXCR4 in a CXCL12-independent manner and worsens tissue remodeling after myocardial infarction. Biochim Biophys Acta Mol basis Dis. 2017;1863:2693–704.CrossRefPubMedGoogle Scholar
Qi YF, Zhang J, Wang L, Shenoy V, Krause E, Oh SP, et al. Angiotensin-converting enzyme 2 inhibits high-mobility group box 1 and attenuates cardiac dysfunction post-myocardial ischemia. J Mol Med. 2016;94:37–49.CrossRefPubMedPubMedCentralGoogle Scholar
Locatelli P, Olea FD, Mendiz O, Salmo F, Fazzi L, Hnatiuk A, et al. An ovine model of postinfarction dilated cardiomyopathy in animals with highly variable coronary anatomy. ILAR J. 2011;52:E16–21.CrossRefPubMedGoogle Scholar
Crottogini A, Meckert PC, Vera Janavel G, Lascano E, Negroni J, Del Valle H, et al. Arteriogenesis induced by intramyocardial vascular endothelial growth factor 165 gene transfer in chronically ischemic pigs. Hum Gene Ther. 2003;14:1307–18.CrossRefPubMedGoogle Scholar
Laguens R, Cabeza Meckert P, Vera Janavel G, De Lorenzi A, Lascano E, Negroni J, et al. Cardiomyocyte hyperplasia after plasmid-mediated vascular endothelial growth factor gene transfer in pigs with chronic myocardial ischemia. J Gene Med. 2004;6:222–7.CrossRefPubMedGoogle Scholar
Locatelli P, Olea FD, De Lorenzi A, Salmo F, Vera Janavel GL, Hnatiuk AP, et al. Reference values for echocardiographic parameters and indexes of left ventricular function in healthy, young adult sheep used in translational research: comparison with standardized values in humans. Int J Clin Exp Med. 2011;4:258–64.PubMedPubMedCentralGoogle Scholar
Gräbner W, Pfitzer P. Number of nuclei in isolated myocardial cells of pigs. Virchows Arch B Cell Pathol. 1974;15:279–94.PubMedGoogle Scholar
Adler CP, Friedburg H, Herget GW, Neuburger M, Schwalb H. Variability of cardiomyocyte DNA content, ploidy level and nuclear number in mammalian hearts. Virchows Arch. 1996;429:159–64.PubMedGoogle Scholar
Limana F, Germani A, Zacheo A, Kajstura J, Di Carlo A, Borsellino G, et al. Exogenous high-mobility group box 1 protein induces myocardial regeneration after infarction via enhanced cardiac C-kit+ cell proliferation and differentiation. Circ Res. 2005;97:e73–83.CrossRefPubMedGoogle Scholar
Nakamura Y, Suzuki S, Shimizu T, Miyata M, Shishido T, Ikeda K, et al. High mobility group box 1 promotes angiogenesis from bone marrow-derived endothelial progenitor cells after myocardial infarction. J Atheroscler Thromb. 2015;22:570–81.CrossRefPubMedGoogle Scholar
Westerhof N, Boer C, Lamberts R, Sipkema P. Cross-talk between cardiac muscle and coronary vasculature. Physiol Rev. 2006;86:1263–308.CrossRefPubMedGoogle Scholar
Opie LH. Oxygen supply: coronary flow. In: Opie LH, editor. The heart. Physiology and metabolism. 2nd ed. New York: Raven Press; 1991. p. 277–81.Google Scholar
Gounis MJ, Spiga MG, Graham RM, Wilson A, Haliko S, Lieber BB, et al. Angiogenesis is confined to the transient period of VEGF expression that follows adenoviral gene delivery to ischemic muscle. Gene Ther. 2005;12:762–71.CrossRefPubMedGoogle Scholar
Olea FD, Vera Janavel G, Cuniberti L, Yannarelli G, Cabeza Meckert P, Cors J, et al. Repeated, but not single, VEGF gene transfer affords protection against ischemic muscle lesions in rabbits with hindlimb ischemia. Gene Ther. 2009;16:716–23.CrossRefPubMedGoogle Scholar
Mitola S, Belleri M, Urbinati C, Coltrini D, Sparatore B, Pedrazzi M, et al. Cutting edge: extracellular high mobility group box-1 protein is a proangiogenic cytokine. J Immunol. 2006;176:12–5.CrossRefPubMedGoogle Scholar
Clayton JA, Chalothorn D, Faber JE. Vascular endothelial growth factor-A specifies formation of native collaterals and regulates collateral growth in ischemia. Circ Res. 2008;103:1027–36.CrossRefPubMedPubMedCentralGoogle Scholar
Stennard FA, Costa MW, Elliott DA, Rankin S, Haast SJ, Lai D, et al. Cardiac T-box factor Tbx20 directly interacts with Nkx2-5, GATA4, and GATA5 in regulation of gene expression in the developing heart. Dev Biol. 2003;262:206–24.CrossRefPubMedGoogle Scholar
Boogerd CJ, Zhu X, Aneas I, Sakabe NJ, Zhang L, Sobreira DR, et al. Tbx20 is required in mid-gestation cardiomyocytes and plays a central role in atrial development. Circ Res. 2018. https://doi.org/10.1161/CIRCRESAHA.118.311339.
Germani A, Limana F, Capogrossi MC. Pivotal advances: high-mobility group box 1 protein - a cytokine with a role in cardiac repair. J Leukoc Biol. 2007;81:41–5.CrossRefPubMedGoogle Scholar
Foglio E, Puddighinu G, Germani A, Russo MA, Limana F. HMGB1 inhibits apoptosis following MI and induces autophagy via mTORC1 inhibition. J Cell Physiol. 2017;232:1135–43.CrossRefPubMedGoogle Scholar
AbdAlla S, Lother H, Abdel-tawab AM, Quitterer U. The angiotensin II AT2 receptor is an AT1 receptor antagonist. J Biol Chem. 2001;276:39721–6.CrossRefPubMedGoogle Scholar