Targeting thrombomodulin to circulating red blood cells augments its protective effects in models of endotoxemia and ischemia-reperfusion injury

Ronald Carnemolla, Carlos H. Villa, Colin F. Greineder, Sergei Zaitsev, Kruti R. Patel, M. Anna Kowalska, Dmitriy N. Atochin, Douglas B. Cines, Don L. Siegel, Charles T. Esmon, Vladimir R. Muzykantov

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8 Citations (Scopus)

Abstract

Endothelial thrombomodulin (TM) regulates coagulation and inflammation via several mechanisms, including production of activated protein C (APC). Recombinant APC and soluble fragments of TM (sTM) have been tested in settings associated with insufficiency of the endogenous TM/APC pathway, such as sepsis. We previously designed a fusion protein of TM [single-chain variable fragment antibody (scFv)/TM] targeted to red bloodcells (RBCs) to improvepharmacokinetics andantithrombotic effectswithout increasingbleeding.Here, scFv/TM was studied in mouse models of systemic inflammation and ischemia-reperfusion injury. Injected concomitantly with or before endotoxin, scFv/TM provided more potent protection against liver injury and release of pathological mediators than sTM, showing similar efficacy at up to 50-fold lower doses. scFv/TM provided protection when injected after endotoxin, whereas sTMdid not, and augmentedAPCproduction by thrombin~50-fold more than sTM. However, scFv/TM injected after endotoxin did not reduce thrombin/antithrombin complexes; nor did antibodies thatblockAPCanticoagulant activity suppress theprophylactic anti-inflammatory effect of scFv/TM. Therefore, similar to endogenous TM, RBC-anchored scFv/TM activates several protective pathways. Finally, scFv/TM wasmore effective at reducing cerebral infarct volume and alleviated neurological deficits than sTM after cerebral ischemia/reperfusion injury. These results indicate that RBC-targeted scFv/TM exerts multifaceted cytoprotective effects and may find utility in systemic and focal inflammatory and ischemic disorders.-Carnemolla, R., Villa, C. H., Greineder, C. F., Zaitseva, S., Patel, K. R., Kowalska, M. A., Atochin, D.N., Cines, D. B., Siegel, D. L., Esmon, C.T., Muzykantov, V.R.Targeting thrombomodulinto circulating red blood cells augments its protective effects inmodels of endotoxemia and ischemia-reperfusion injury.

Original languageEnglish
Pages (from-to)761-770
Number of pages10
JournalFASEB Journal
Volume31
Issue number2
DOIs
Publication statusPublished - 1 Feb 2017
Externally publishedYes

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Keywords

  • Coagulation
  • Drug delivery
  • Inflammation
  • Pharmacokinetics
  • Sepsis

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Carnemolla, R., Villa, C. H., Greineder, C. F., Zaitsev, S., Patel, K. R., Kowalska, M. A., Atochin, D. N., Cines, D. B., Siegel, D. L., Esmon, C. T., & Muzykantov, V. R. (2017). Targeting thrombomodulin to circulating red blood cells augments its protective effects in models of endotoxemia and ischemia-reperfusion injury. FASEB Journal, 31(2), 761-770. https://doi.org/10.1096/fj.201600912R