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MMP10 Promotes Efficient Thrombolysis After Ischemic Stroke in Mice with Induced Diabetes

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Abstract

Diabetes is an important risk factor for ischemic stroke (IS). Tissue-type plasminogen activator (tPA) has been associated with less successful revascularization and poor functional outcome in diabetes. We assessed whether a new thrombolytic strategy based on MMP10 was more effective than tPA in a murine IS model of streptozotocin (STZ)-induced diabetes. Wild-type mice were administered a single dose of streptozotocin (STZ) (180 mg/kg) to develop STZ-induced diabetes mellitus. Two weeks later, IS was induced by thrombin injection into the middle cerebral artery and the effect of recombinant MMP10 (6.5 μg/kg), tPA (10 mg/kg) or tPA/MMP10 on brain damage and functional outcome were analysed. Motor activity was assessed using the open field test. Additionally, we studied plasminogen activator inhibitor-1 (PAI-1) and thrombin-antithrombin complex levels (TAT) by ELISA and oxidative stress and blood-brain barrier (BBB) integrity by immunohistochemistry and western blot. MMP10 treatment was more effective at reducing infarct size and neurodegeneration than tPA 24 h and 3 days after IS in diabetic mice. Locomotor activity was impaired by hyperglycemia and ischemic injury, but not by the thrombolytic treatments. Additionally, TAT, oxidative stress and BBB permeability were reduced by MMP10 treatment, whereas brain bleeding or PAI-1 expression did not differ between treatments. Thrombolytic treatment with MMP10 was more effective than tPA at reducing stroke and neurodegeneration in a diabetic murine model of IS, without increasing haemorrhage. Thus, we propose MMP10 as a potential candidate for the clinical treatment of IS in diabetic patients.

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References

  1. Kearney K, Tomlinson D, Smith K, Ajjan R. Hypofibrinolysis in diabetes: a therapeutic target for the reduction of cardiovascular risk. Cardiovasc Diabetol. 2017;16(1):34. https://doi.org/10.1186/s12933-017-0515-9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Tsujimoto T, Sugiyama T, Shapiro MF, Noda M, Kajio H. Risk of cardiovascular events in patients with diabetes mellitus on beta-blockers. Hypertension. 2017;70(1):103–10. https://doi.org/10.1161/HYPERTENSIONAHA.117.09259.

    Article  CAS  PubMed  Google Scholar 

  3. Papatheodorou K, Banach M, Edmonds M, Papanas N, Papazoglou D. Complications of diabetes. J Diabetes Res. 2015;2015:189525. https://doi.org/10.1155/2015/189525.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Kruyt ND, Biessels GJ, Devries JH, Roos YB. Hyperglycemia in acute ischemic stroke: pathophysiology and clinical management. Nat Rev Neurol. 2010;6(3):145–55. https://doi.org/10.1038/nrneurol.2009.231.

    Article  CAS  PubMed  Google Scholar 

  5. Lee SJ, Hong JM, Lee SE, Kang DR, Ovbiagele B, Demchuk AM, et al. Association of fibrinogen level with early neurological deterioration among acute ischemic stroke patients with diabetes. BMC Neurol. 2017;17(1):101. https://doi.org/10.1186/s12883-017-0865-7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Orset C, Macrez R, Young AR, Panthou D, Angles-Cano E, Maubert E, et al. Mouse model of in situ thromboembolic stroke and reperfusion. Stroke. 2007;38(10):2771–8. https://doi.org/10.1161/STROKEAHA.107.487520.

    Article  PubMed  Google Scholar 

  7. Simao F, Ustunkaya T, Clermont AC, Feener EP. Plasma kallikrein mediates brain hemorrhage and edema caused by tissue plasminogen activator therapy in mice after stroke. Blood. 2017;129(16):2280–90. https://doi.org/10.1182/blood-2016-09-740670.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Dunn EJ, Philippou H, Ariens RA, Grant PJ. Molecular mechanisms involved in the resistance of fibrin to clot lysis by plasmin in subjects with type 2 diabetes mellitus. Diabetologia. 2006;49(5):1071–80. https://doi.org/10.1007/s00125-006-0197-4.

    Article  CAS  PubMed  Google Scholar 

  9. Kamada H, Yu F, Nito C, Chan PH. Influence of hyperglycemia on oxidative stress and matrix metalloproteinase-9 activation after focal cerebral ischemia/reperfusion in rats: relation to blood-brain barrier dysfunction. Stroke. 2007;38(3):1044–9. https://doi.org/10.1161/01.STR.0000258041.75739.cb.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Vila N, Castillo J, Davalos A, Chamorro A. Proinflammatory cytokines and early neurological worsening in ischemic stroke. Stroke. 2000;31(10):2325–9.

    Article  CAS  PubMed  Google Scholar 

  11. Xiang FL, Lu X, Strutt B, Hill DJ, Feng Q. NOX2 deficiency protects against streptozotocin-induced beta-cell destruction and development of diabetes in mice. Diabetes. 2010;59(10):2603–11. https://doi.org/10.2337/db09-1562.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Ceriello A, Novials A, Ortega E, Pujadas G, La Sala L, Testa R, et al. Hyperglycemia following recovery from hypoglycemia worsens endothelial damage and thrombosis activation in type 1 diabetes and in healthy controls. Nutr Metab Cardiovasc Dis. 2014;24(2):116–23. https://doi.org/10.1016/j.numecd.2013.05.003.

    Article  CAS  PubMed  Google Scholar 

  13. Fernandez-Cadenas I, Mendioroz M, Munuera J, Alvarez-Sabin J, Rovira A, Quiroga A, et al. Lower concentrations of thrombin-antithrombin complex (TAT) correlate to higher recanalisation rates among ischaemic stroke patients treated with t-PA. Thromb Haemost. 2009;102(4):759–64. https://doi.org/10.1160/TH08-06-0398.

    Article  CAS  PubMed  Google Scholar 

  14. Kota SK, Meher LK, Jammula S, Kota SK, Krishna SV, Modi KD. Aberrant angiogenesis: the gateway to diabetic complications. Indian J Endocrinol Metab. 2012;16(6):918–30. https://doi.org/10.4103/2230-8210.102992.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Coll B, Rodriguez JA, Craver L, Orbe J, Martinez-Alonso M, Ortiz A, et al. Serum levels of matrix metalloproteinase-10 are associated with the severity of atherosclerosis in patients with chronic kidney disease. Kidney Int. 2010;78(12):1275–80. https://doi.org/10.1038/ki.2010.329.

    Article  CAS  PubMed  Google Scholar 

  16. Cunningham LA, Wetzel M, Rosenberg GA. Multiple roles for MMPs and TIMPs in cerebral ischemia. Glia. 2005;50(4):329–39. https://doi.org/10.1002/glia.20169.

    Article  PubMed  Google Scholar 

  17. Toni M, Hermida J, Goni MJ, Fernandez P, Parks WC, Toledo E, et al. Matrix metalloproteinase-10 plays an active role in microvascular complications in type 1 diabetic patients. Diabetologia. 2013;56(12):2743–52. https://doi.org/10.1007/s00125-013-3052-4.

    Article  CAS  PubMed  Google Scholar 

  18. Rodriguez JA, Orbe J, Martinez de Lizarrondo S, Calvayrac O, Rodriguez C, Martinez-Gonzalez J, et al. Metalloproteinases and atherothrombosis: MMP-10 mediates vascular remodeling promoted by inflammatory stimuli. Front Biosci. 2008;13:2916–21.

    Article  CAS  PubMed  Google Scholar 

  19. Orbe J, Barrenetxe J, Rodriguez JA, Vivien D, Orset C, Parks WC, et al. Matrix metalloproteinase-10 effectively reduces infarct size in experimental stroke by enhancing fibrinolysis via a thrombin-activatable fibrinolysis inhibitor-mediated mechanism. Circulation. 2011;124(25):2909–19. https://doi.org/10.1161/CIRCULATIONAHA.111.047100.

    Article  CAS  PubMed  Google Scholar 

  20. Rodriguez JA, Sobrino T, Lopez-Arias E, Ugarte A, Sanchez-Arias JA, Vieites-Prado A, et al. CM352 Reduces brain damage and improves functional recovery in a rat model of intracerebral hemorrhage. J Am Heart Assoc. 2017;6(6) https://doi.org/10.1161/JAHA.117.006042.

  21. Roncal C, Martinez de Lizarrondo S, Salicio A, Chevilley A, Rodriguez JA, Rosell A, et al. New thrombolytic strategy providing neuroprotection in experimental ischemic stroke: MMP10 alone or in combination with tissue-type plasminogen activator. Cardiovasc Res. 2017;113(10):1219–29. https://doi.org/10.1093/cvr/cvx069.

    Article  CAS  PubMed  Google Scholar 

  22. Rosell A, Ortega-Aznar A, Alvarez-Sabin J, Fernandez-Cadenas I, Ribo M, Molina CA, et al. Increased brain expression of matrix metalloproteinase-9 after ischemic and hemorrhagic human stroke. Stroke. 2006;37(6):1399–406. https://doi.org/10.1161/01.STR.0000223001.06264.af.

    Article  CAS  PubMed  Google Scholar 

  23. Kahles T, Brandes RP. Which NADPH oxidase isoform is relevant for ischemic stroke? The case for nox 2. Antioxid Redox Signal. 2013;18(12):1400–17. https://doi.org/10.1089/ars.2012.4721.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Jauch EC, Saver JL, Adams HP Jr, Bruno A, Connors JJ, Demaerschalk BM, et al. Guidelines for the early management of patients with acute ischemic stroke: a guideline for healthcare professionals from the American Heart Association/American Stroke Association. Stroke. 2013;44(3):870–947. https://doi.org/10.1161/STR.0b013e318284056a.

    Article  PubMed  Google Scholar 

  25. Hafez S, Coucha M, Bruno A, Fagan SC, Ergul A. Hyperglycemia, acute ischemic stroke, and thrombolytic therapy. Transl Stroke Res. 2014;5(4):442–53. https://doi.org/10.1007/s12975-014-0336-z.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Bruno A, Liebeskind D, Hao Q, Raychev R, Investigators US. Diabetes mellitus, acute hyperglycemia, and ischemic stroke. Curr Treat Options Neurol. 2010;12(6):492–503. https://doi.org/10.1007/s11940-010-0093-6.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Fan X, Qiu J, Yu Z, Dai H, Singhal AB, Lo EH, et al. A rat model of studying tissue-type plasminogen activator thrombolysis in ischemic stroke with diabetes. Stroke. 2012;43(2):567–70. https://doi.org/10.1161/STROKEAHA.111.635250.

    Article  CAS  PubMed  Google Scholar 

  28. European Stroke Organisation Executive C, Committee ESOW. Guidelines for management of ischaemic stroke and transient ischaemic attack 2008. Cerebrovasc Dis. 2008;25(5):457–507. https://doi.org/10.1159/000131083.

    Article  Google Scholar 

  29. Hawkins BT, Davis TP. The blood-brain barrier/neurovascular unit in health and disease. Pharmacol Rev. 2005;57(2):173–85. https://doi.org/10.1124/pr.57.2.4.

    Article  CAS  PubMed  Google Scholar 

  30. Abdelsaid M, Prakash R, Li W, Coucha M, Hafez S, Johnson MH, et al. Metformin treatment in the period after stroke prevents nitrative stress and restores angiogenic signaling in the brain in diabetes. Diabetes. 2015;64(5):1804–17. https://doi.org/10.2337/db14-1423.

    Article  CAS  PubMed  Google Scholar 

  31. Rosell A, Lo EH. Multiphasic roles for matrix metalloproteinases after stroke. Curr Opin Pharmacol. 2008;8(1):82–9. https://doi.org/10.1016/j.coph.2007.12.001.

    Article  CAS  PubMed  Google Scholar 

  32. Ning R, Chopp M, Yan T, Zacharek A, Zhang C, Roberts C, et al. Tissue plasminogen activator treatment of stroke in type-1 diabetes rats. Neuroscience. 2012;222:326–32. https://doi.org/10.1016/j.neuroscience.2012.07.018.

    Article  CAS  PubMed  Google Scholar 

  33. Orset C, Haelewyn B, Allan SM, Ansar S, Campos F, Cho TH, et al. Efficacy of alteplase in a mouse model of acute ischemic stroke: a retrospective pooled analysis. Stroke. 2016;47(5):1312–8. https://doi.org/10.1161/STROKEAHA.116.012238.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Montaner J, Molina CA, Monasterio J, Abilleira S, Arenillas JF, Ribo M, et al. Matrix metalloproteinase-9 pretreatment level predicts intracranial hemorrhagic complications after thrombolysis in human stroke. Circulation. 2003;107(4):598–603.

    Article  CAS  PubMed  Google Scholar 

  35. Inzitari D, Giusti B, Nencini P, Gori AM, Nesi M, Palumbo V, et al. MMP9 variation after thrombolysis is associated with hemorrhagic transformation of lesion and death. Stroke. 2013;44(10):2901–3. https://doi.org/10.1161/Strokeaha.113.002274.

    Article  CAS  PubMed  Google Scholar 

  36. Kowluru RA, Mohammad G, dos Santos JM, Zhong Q. Abrogation of MMP-9 gene protects against the development of retinopathy in diabetic mice by preventing mitochondrial damage. Diabetes. 2011;60(11):3023–33. https://doi.org/10.2337/db11-0816.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Desilles JP, Syvannarath V, Ollivier V, Journe C, Delbosc S, Ducroux C, et al. Exacerbation of thromboinflammation by hyperglycemia precipitates cerebral infarct growth and hemorrhagic transformation. Stroke. 2017;48(7):1932–40. https://doi.org/10.1161/STROKEAHA.117.017080.

    Article  PubMed  Google Scholar 

  38. Kahles T, Brandes RP. NADPH oxidases as therapeutic targets in ischemic stroke. Cell Mol Life Sci. 2012;69(14):2345–63. https://doi.org/10.1007/s00018-012-1011-8.

    Article  CAS  PubMed  Google Scholar 

  39. Liu J, Jin X, Liu KJ, Liu W. Matrix metalloproteinase-2-mediated occludin degradation and caveolin-1-mediated claudin-5 redistribution contribute to blood-brain barrier damage in early ischemic stroke stage. J Neurosci. 2012;32(9):3044–57. https://doi.org/10.1523/JNEUROSCI.6409-11.2012.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Jiao H, Wang Z, Liu Y, Wang P, Xue Y. Specific role of tight junction proteins claudin-5, occludin, and ZO-1 of the blood-brain barrier in a focal cerebral ischemic insult. J Mol Neurosci. 2011;44(2):130–9. https://doi.org/10.1007/s12031-011-9496-4.

    Article  CAS  PubMed  Google Scholar 

  41. Borsello T, Clarke PG, Hirt L, Vercelli A, Repici M, Schorderet DF, et al. A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia. Nat Med. 2003;9(9):1180–6. https://doi.org/10.1038/nm911.

    Article  CAS  PubMed  Google Scholar 

  42. Ou Z, Tao MX, Gao Q, Zhang XL, Yang Y, Zhou JS, et al. Up-regulation of angiotensin-converting enzyme in response to acute ischemic stroke via ERK/NF-kappaB pathway in spontaneously hypertensive rats. Oncotarget. 2017;8(57):97041–51. https://doi.org/10.18632/oncotarget.21156.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Jung JE, Kim GS, Chen H, Maier CM, Narasimhan P, Song YS, et al. Reperfusion and neurovascular dysfunction in stroke: from basic mechanisms to potential strategies for neuroprotection. Mol Neurobiol. 2010;41(2–3):172–9. https://doi.org/10.1007/s12035-010-8102-z.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Newsholme P, Cruzat VF, Keane KN, Carlessi R, de Bittencourt PI, Jr. Molecular mechanisms of ROS production and oxidative stress in diabetes. The Biochemical Journal. 2016;473(24):4527–50. https://doi.org/10.1042/BCJ20160503C.

    Article  CAS  PubMed  Google Scholar 

  45. Suh SW, Shin BS, Ma H, Van Hoecke M, Brennan AM, Yenari MA, et al. Glucose and NADPH oxidase drive neuronal superoxide formation in stroke. Ann Neurol. 2008;64(6):654–63. https://doi.org/10.1002/ana.21511.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Liu W, Sood R, Chen Q, Sakoglu U, Hendren J, Cetin O, et al. Normobaric hyperoxia inhibits NADPH oxidase-mediated matrix metalloproteinase-9 induction in cerebral microvessels in experimental stroke. J Neurochem. 2008;107(5):1196–205. https://doi.org/10.1111/j.1471-4159.2008.05664.x.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Miller AA, Dusting GJ, Roulston CL, Sobey CG. NADPH-oxidase activity is elevated in penumbral and non-ischemic cerebral arteries following stroke. Brain Res. 2006;1111(1):111–6. https://doi.org/10.1016/j.brainres.2006.06.082.

    Article  CAS  PubMed  Google Scholar 

  48. Ribo M, Montaner J, Molina CA, Arenillas JF, Santamarina E, Alvarez-Sabin J. Admission fibrinolytic profile predicts clot lysis resistance in stroke patients treated with tissue plasminogen activator. Thromb Haemost. 2004;91(6):1146–51. https://doi.org/10.1160/TH04-02-0097.

    Article  CAS  PubMed  Google Scholar 

  49. Domingueti CP, Dusse LM, Carvalho M, de Sousa LP, Gomes KB, Fernandes AP. Diabetes mellitus: the linkage between oxidative stress, inflammation, hypercoagulability and vascular complications. J Diabetes Complicat. 2016;30(4):738–45. https://doi.org/10.1016/j.jdiacomp.2015.12.018.

    Article  Google Scholar 

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Acknowledgements

We thank Lara Montori for the help with the experimental work and Rosa Tordera and Mikel Aleixo from Pharmacology and Toxicology Department, University of Navarra for their technical assistance with behavioural tests.

Funding

This work was supported by the Ministerio de Economía y Competitividad, Instituto de Salud Carlos III [FIS PI15/01807] and Federal Ministry of Education and Research (FEDER) funds; grants from the Spanish Society of Thrombosis and Haemostasis (SETH), Navarra Government (02/2015), Patrimonio Praga (Mexico) and Virto Group (Navarra, Spain); and PhD scholarship from the Asociación de Amigos de la Universidad de Navarra (ADA).

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Authors and Affiliations

  1. Atherothrombosis Research Laboratory, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain

    Manuel Navarro-Oviedo, Carmen Roncal, Agustina Salicio, Miriam Belzunce, Jose A. Rodríguez & Josune Orbe

  2. CIBER Cardiovascular (CIBERCV), Ministry of Economy and Competitiveness, ISCIII, Madrid, Spain

    Carmen Roncal, Agustina Salicio, Jose A. Rodríguez, Jose A. Páramo & Josune Orbe

  3. IdiSNA, Instituto de Investigación Sanitaria de Navarra, Pamplona, Spain

    Carmen Roncal, Estefanía Toledo, Jose A. Rodríguez, Jose A. Páramo, Roberto Muñoz & Josune Orbe

  4. Small Molecule Discovery Platform, Molecular Therapeutics Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain

    Obdulia Rabal

  5. Department of Preventive Medicine and Public Health, School of Medicine, University of Navarra, Pamplona, Spain

    Estefanía Toledo

  6. CIBER Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Ministry of Economy and Competitiveness, ISCIII, Pamplona, Spain

    Estefanía Toledo

  7. Neurology Department, Complejo Hospitalario de Navarra, Pamplona, Spain

    Beatriz Zandio & Roberto Muñoz

  8. Haematology Service, Clínica Universidad de Navarra, University of Navarra, Pamplona, Spain

    Jose A. Páramo

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  1. Manuel Navarro-Oviedo
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  2. Carmen Roncal
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Contributions

MNO participated in the design of the project, experimental work, statistical analysis, and wrote, reviewed, and edited the manuscript; CR participated in the design of the project and reviewed the manuscript. AS was in charge of the animal experiments; MB performed histological studies; OR participated in the design of the project and reviewed the manuscript; ET participated in the statistical analysis and reviewed the manuscript; BZ participated in the design of the project and reviewed the manuscript; JAR participated in the design of the project and reviewed the manuscript; JAP participated in the design of the project, supervised the work, and edited and reviewed the manuscript; RM participated in the design of the project and edited and reviewed the manuscript; JO was in charge of the whole project design, supervised the work, and wrote, edited, and reviewed this manuscript.

Corresponding author

Correspondence to Josune Orbe.

Ethics declarations

Experiments were performed in accordance with European Communities Council Directive guidelines (2010/63/EU) for the care and use of laboratory animals and were approved by the University of Navarra Animal Research Review Committee.

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

All animal experiments were performed in accordance with European Communities Council Directive guidelines (2010/63/EU) for the care and use of laboratory animals and were approved by the University of Navarra Animal Research Review Committee.

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Navarro-Oviedo, M., Roncal, C., Salicio, A. et al. MMP10 Promotes Efficient Thrombolysis After Ischemic Stroke in Mice with Induced Diabetes. Transl. Stroke Res. 10, 389–401 (2019). https://doi.org/10.1007/s12975-018-0652-9

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