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 language | English |
|---|---|
| Pages (from-to) | 761-770 |
| Number of pages | 10 |
| Journal | FASEB Journal |
| Volume | 31 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 1 Feb 2017 |
| Externally published | Yes |
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Keywords
- Coagulation
- Drug delivery
- Inflammation
- Pharmacokinetics
- Sepsis
ASJC Scopus subject areas
- Biotechnology
- Biochemistry
- Molecular Biology
- Genetics
Cite this
Targeting thrombomodulin to circulating red blood cells augments its protective effects in models of endotoxemia and ischemia-reperfusion injury. / Carnemolla, Ronald; Villa, Carlos H.; Greineder, Colin F.; Zaitsev, Sergei; Patel, Kruti R.; Kowalska, M. Anna; Atochin, Dmitriy N.; Cines, Douglas B.; Siegel, Don L.; Esmon, Charles T.; Muzykantov, Vladimir R.
In: FASEB Journal, Vol. 31, No. 2, 01.02.2017, p. 761-770.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Targeting thrombomodulin to circulating red blood cells augments its protective effects in models of endotoxemia and ischemia-reperfusion injury
AU - Carnemolla, Ronald
AU - Villa, Carlos H.
AU - Greineder, Colin F.
AU - Zaitsev, Sergei
AU - Patel, Kruti R.
AU - Kowalska, M. Anna
AU - Atochin, Dmitriy N.
AU - Cines, Douglas B.
AU - Siegel, Don L.
AU - Esmon, Charles T.
AU - Muzykantov, Vladimir R.
PY - 2017/2/1
Y1 - 2017/2/1
N2 - 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.
AB - 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.
KW - Coagulation
KW - Drug delivery
KW - Inflammation
KW - Pharmacokinetics
KW - Sepsis
UR - http://www.scopus.com/inward/record.url?scp=85011255374&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85011255374&partnerID=8YFLogxK
U2 - 10.1096/fj.201600912R
DO - 10.1096/fj.201600912R
M3 - Article
VL - 31
SP - 761
EP - 770
JO - FASEB Journal
JF - FASEB Journal
SN - 0892-6638
IS - 2
ER -