Harnessing Micrometer-scale TPA Beads for High Plasmin Generation and Accelerated Fibrinolysis

Overview

Journal bioRxiv

Date 2024 Nov 22

PMID 39574757

Authors

Matthew J Osmond

Fabrice Dabertrand

Nidia Quillinan

Enming J Su

Daniel A Lawrence

David W M Marr

Keith B Neeves

Affiliations

Soon will be listed here.

Abstract

Rapid restoration of blood flow is critical in treating acute ischemic stroke. Current fibrinolytic therapies using tissue plasminogen activator (tPA) are limited by low recanalization rates and risks of off-target bleeding. Here, we present a strategy using tPA immobilized on micrometer-scale beads to enhance local plasmin generation. We synthesized tPA-functionalized beads of varying sizes (0.1 μm and 1.0 μm) and evaluated their efficacy. assays demonstrated that 1.0 μm tPA-beads generated higher plasmin generation compared to free tPA and 0.1 μm beads, overcoming antiplasmin inhibition and promoting a self-propagating wave of fibrinolysis. In a murine model of acute ischemic stroke, intravenous administration of 1.0 μm tPA-beads at doses nearly two orders of magnitude lower than the standard free tPA dose led to rapid and near-complete thrombus removal within minutes. This approach addresses kinetic and transport limitations of current therapies and may reduce the risk of hemorrhagic complications.

References

Huang Y, Gu B, Salles-Crawley I, Taylor K, Yu L, Ren J . Fibrinogen-mimicking, multiarm nanovesicles for human thrombus-specific delivery of tissue plasminogen activator and targeted thrombolytic therapy. Sci Adv. 2021; 7(23). PMC: 8172176. DOI: 10.1126/sciadv.abf9033. View

Wu J, Siddiqui K, Diamond S . Transport phenomena and clot dissolving therapy: an experimental investigation of diffusion-controlled and permeation-enhanced fibrinolysis. Thromb Haemost. 1994; 72(1):105-12. View

Roth D, Piekarek M, Paulsson M, Christ H, Bloch W, Krieg T . Plasmin modulates vascular endothelial growth factor-A-mediated angiogenesis during wound repair. Am J Pathol. 2006; 168(2):670-84. PMC: 1606492. DOI: 10.2353/ajpath.2006.050372. View

Diamond S . Engineering design of optimal strategies for blood clot dissolution. Annu Rev Biomed Eng. 2001; 1:427-62. DOI: 10.1146/annurev.bioeng.1.1.427. View

Su E, Fredriksson L, Geyer M, Folestad E, Cale J, Andrae J . Activation of PDGF-CC by tissue plasminogen activator impairs blood-brain barrier integrity during ischemic stroke. Nat Med. 2008; 14(7):731-7. PMC: 2811427. DOI: 10.1038/nm1787. View

Modi N, Fox N, Clow F, Tanswell P, Cannon C, Van de Werf F . Pharmacokinetics and pharmacodynamics of tenecteplase: results from a phase II study in patients with acute myocardial infarction. J Clin Pharmacol. 2000; 40(5):508-15. DOI: 10.1177/00912700022009125. View

Kim P, Stewart R, Lipson S, Nesheim M . The relative kinetics of clotting and lysis provide a biochemical rationale for the correlation between elevated fibrinogen and cardiovascular disease. J Thromb Haemost. 2007; 5(6):1250-6. DOI: 10.1111/j.1538-7836.2007.02426.x. View

Kolev K, Longstaff C . Bleeding related to disturbed fibrinolysis. Br J Haematol. 2016; 175(1):12-23. PMC: 5096260. DOI: 10.1111/bjh.14255. View

Matosevic B, Knoflach M, Werner P, Pechlaner R, Zangerle A, Ruecker M . Fibrinogen degradation coagulopathy and bleeding complications after stroke thrombolysis. Neurology. 2013; 80(13):1216-24. DOI: 10.1212/WNL.0b013e3182897015. View

10.

Chen J, Yang P, Ma Y, Tu S, Lu Y . Targeted delivery of tissue plasminogen activator by binding to silica-coated magnetic nanoparticle. Int J Nanomedicine. 2012; 7:5137-49. PMC: 3463402. DOI: 10.2147/IJN.S36197. View

11.

Marder V, Novokhatny V . Direct fibrinolytic agents: biochemical attributes, preclinical foundation and clinical potential. J Thromb Haemost. 2009; 8(3):433-44. DOI: 10.1111/j.1538-7836.2009.03701.x. View

12.

Lijnen H, Collen D . Mechanisms of physiological fibrinolysis. Baillieres Clin Haematol. 1995; 8(2):277-90. DOI: 10.1016/s0950-3536(05)80268-9. View

13.

Tasci T, Disharoon D, Schoeman R, Rana K, Herson P, Marr D . Enhanced Fibrinolysis with Magnetically Powered Colloidal Microwheels. Small. 2017; 13(36). PMC: 7927958. DOI: 10.1002/smll.201700954. View

14.

Klegerman M, Zou Y, McPherson D . Fibrin targeting of echogenic liposomes with inactivated tissue plasminogen activator. J Liposome Res. 2008; 18(2):95-112. DOI: 10.1080/08982100802118482. View

15.

Zaidat O, Suarez J, Sunshine J, Tarr R, Alexander M, Smith T . Thrombolytic therapy of acute ischemic stroke: correlation of angiographic recanalization with clinical outcome. AJNR Am J Neuroradiol. 2005; 26(4):880-4. PMC: 7977112. View

16.

Wu J, Diamond S . Tissue plasminogen activator (tPA) inhibits plasmin degradation of fibrin. A mechanism that slows tPA-mediated fibrinolysis but does not require alpha 2-antiplasmin or leakage of intrinsic plasminogen. J Clin Invest. 1995; 95(6):2483-90. PMC: 295930. DOI: 10.1172/JCI117949. View

17.

Wang Y, Tsirka S, Strickland S, Stieg P, Soriano S, Lipton S . Tissue plasminogen activator (tPA) increases neuronal damage after focal cerebral ischemia in wild-type and tPA-deficient mice. Nat Med. 1998; 4(2):228-31. DOI: 10.1038/nm0298-228. View

18.

Sun Y, Kuo Y, Chen H, Short-Miller J, Smucker M, Kuan C . A murine photothrombotic stroke model with an increased fibrin content and improved responses to tPA-lytic treatment. Blood Adv. 2020; 4(7):1222-1231. PMC: 7160278. DOI: 10.1182/bloodadvances.2019000782. View

19.

Thomalla G, Schwark C, Sobesky J, Bluhmki E, Fiebach J, Fiehler J . Outcome and symptomatic bleeding complications of intravenous thrombolysis within 6 hours in MRI-selected stroke patients: comparison of a German multicenter study with the pooled data of ATLANTIS, ECASS, and NINDS tPA trials. Stroke. 2006; 37(3):852-8. DOI: 10.1161/01.STR.0000204120.79399.72. View

20.

Nagai N, Suzuki Y, Van Hoef B, Lijnen H, Collen D . Effects of plasminogen activator inhibitor-1 on ischemic brain injury in permanent and thrombotic middle cerebral artery occlusion models in mice. J Thromb Haemost. 2005; 3(7):1379-84. DOI: 10.1111/j.1538-7836.2005.01466.x. View