Anticoagulation Targeting Membrane-Bound Anionic Phospholipids Improved Outcomes of Traumatic Brain Injury in Mice

Blood ◽  
2021 ◽  
Author(s):  
Xinlong Dong ◽  
Wei Liu ◽  
Yu Shen ◽  
Katie L Houck ◽  
Mengchen Yang ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Endothelial Cells ◽  
Brain Injury ◽  
Extracellular Vesicles ◽  
Annexin V ◽  
Hypercoagulable State ◽  
Anionic Phospholipid ◽  
Anionic Phospholipids ◽  
Fluid Percussion Injury ◽  
Tissue Edema

Severe traumatic brain injury (TBI) often causes an acute systemic hypercoagulable state that rapidly develops into consumptive coagulopathy. We have recently demonstrated that TBI-induced coagulopathy (TBI-IC) is initiated and disseminated by brain-derived extracellular vesicles (BDEVs) and propagated by extracellular vesicles (EVs) from endothelial cells and platelets. Here, we present results from a study designed to test the hypothesis that anticoagulation targeting anionic phospholipid-expressing EVs prevents TBI-IC and improves the outcomes of mice subjected to severe TBI. We evaluated the effects of a fusion protein (ANV-6L15) for improving the outcomes of TBI. ANV-6L15 combines the phosphatidylserine (PS)-binding annexin V with a peptide anticoagulant modified to preferentially target extrinsic coagulation. We found that ANV-6L15 reduced intracranial hematoma by 70.2%, improved neurological function, and reduced death by 56.8% in mice subjected to fluid percussion injury at 1.9 atm. It protected the TBI mice by preventing vascular leakage, tissue edema, and the TBI-induced hypercoagulable state. We further showed that the extrinsic tenase complex was formed on the surfaces of circulating EVs, with the highest level found on BDEVs. Phospholipidomic analysis detected the highest levels of PS on BDEVs, as compared to EVs from endothelial cells and platelets (79.1, 15.2, and 3.5 nM/mg of protein, respectively). These findings demonstrate that TBI-IC results from a trauma-induced hypercoagulable state and may be treated by anticoagulation targeting on the anionic phospholipid-expressing membrane of EVs from the brain and other cells.

scholarly journals Neurodegeneration and Sensorimotor Deficits in the Mouse Model of Traumatic Brain Injury

2017 ◽  
Author(s):  
Saurav Bhowmick ◽  
Veera D'Mello ◽  
Nizmi Ponery ◽  
P.M. Abdul-Muneer
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Caspase 3 ◽  
Annexin V ◽  
Screen Test ◽  
In Vivo Model ◽  
Tunel Staining ◽  
Sensorimotor Deficits ◽  
Fluid Percussion Injury

Traumatic brain injury (TBI) can result in persistent sensorimotor and cognitive deficits, which occur through a cascade of deleterious pathophysiological events over time. In this study, we investigated the hypothesis that neurodegeneration caused by TBI leads to impairments in sensorimotor function. TBI induces the activation of the caspase-3 enzyme, which triggers cell apoptosis in an in vivo model of fluid percussion injury (FPI). We analyzed caspase-3 mediated apoptosis by TUNEL staining and PARP and annexin V western blotting. We correlated the neurodegeneration with sensorimotor deficits by conducting the animal behavioral tests including grid walk, balance beam, inverted screen test, and climb test. Our study demonstrated that the excess cell death or neurodegeneration correlated with the neuronal dysfunction and sensorimotor impairments associated with TBI.

scholarly journals Rapid Accumulation of Endogenous Tau Oligomers in a Rat Model of Traumatic Brain Injury

2013 ◽  
Vol 288 (23) ◽  
pp. 17042-17050 ◽  
Author(s):  
Bridget E. Hawkins ◽  
Shashirekha Krishnamurthy ◽  
Diana L. Castillo-Carranza ◽  
Urmi Sengupta ◽  
Donald S. Prough ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
The United States ◽  
Tau Proteins ◽  
Diagnostic Tools ◽  
Phosphorylated Tau ◽  
Fluid Percussion Injury ◽  
Rapid Accumulation ◽  
Severe Tbi ◽  
Effective Treatments

Traumatic brain injury (TBI) is a serious problem that affects millions of people in the United States alone. Multiple concussions or even a single moderate to severe TBI can also predispose individuals to develop a pathologically distinct form of tauopathy-related dementia at an early age. No effective treatments are currently available for TBI or TBI-related dementia; moreover, only recently has insight been gained regarding the mechanisms behind their connection. Here, we used antibodies to detect oligomeric and phosphorylated Tau proteins in a non-transgenic rodent model of parasagittal fluid percussion injury. Oligomeric and phosphorylated Tau proteins were detected 4 and 24 h and 2 weeks post-TBI in injured, but not sham control rats. These findings suggest that diagnostic tools and therapeutics that target only toxic forms of Tau may provide earlier detection and safe, more effective treatments for tauopathies associated with repetitive neurotrauma.

scholarly journals Potential biomarkers to detect traumatic brain injury by the profiling of salivary extracellular vesicles

2019 ◽  
Vol 234 (8) ◽  
pp. 14377-14388 ◽  
Author(s):  
Yan Cheng ◽  
Mandy Pereira ◽  
Neha Raukar ◽  
John L. Reagan ◽  
Mathew Queseneberry ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Extracellular Vesicles ◽  
Potential Biomarkers

Ligand-regulated secretion of recombinant annexin V from cultured thyroid epithelial cells

2002 ◽  
Vol 282 (6) ◽  
pp. C1313-C1321 ◽  
Author(s):  
Xiuqiong Wang ◽  
Marcia A. Kaetzel ◽  
Sung E. Yoo ◽  
Paul S. Kim ◽  
John R. Dedman
Keyword(s):  
Mammalian Cells ◽  
Secretory Pathway ◽  
External Surface ◽  
Annexin V ◽  
Therapeutic Agents ◽  
Anionic Phospholipid ◽  
Thyroid Cells ◽  
Tissue Ischemia ◽  
Anionic Phospholipids ◽  
Activated Platelets

The exposure of anionic phospholipids on the external surface of injured endothelial cells and activated platelets is a primary biological signal to initiate blood coagulation. Disease conditions that promote the formation of ectopic thrombi result in tissue ischemia. Annexins, Ca2+-dependent anionic phospholipid binding proteins, are potential therapeutic agents for the inhibition of coagulation. We have designed a transgene that targets secretion of annexin V from cultured thyroid cells under the control of doxycycline. Our results indicate that annexin V in the endoplasmic reticulum (ER)/Golgi lumen does not affect the synthesis, processing, and secretion of thyroglobulin. ER luminal Ca2+was moderately increased and can be released by inositol 1,4,5-trisphosphate. Our study demonstrates that targeting and secretion of annexin V through the secretory pathway of mammalian cells does not adversely affect cellular function. Regulated synthesis and release of annexin V may exert anticoagulatory and anti-inflammatory effects systemically and may prove useful in further developing therapeutic strategies for conditions including antiphospholipid syndrome.

scholarly journals Oyxsterols induce membrane procoagulant activity in monocytic THP-1 cells

1996 ◽  
Vol 314 (3) ◽  
pp. 1027-1033 ◽  
Author(s):  
Karine AUPEIX ◽  
Florence TOTI ◽  
Nathalie SATTA ◽  
Pierre BISCHOFF ◽  
Jean-Marie FREYSSINET
Keyword(s):  
Tissue Factor ◽  
Annexin V ◽  
Antigen Expression ◽  
Resting Cells ◽  
Dependent Manner ◽  
Anionic Phospholipid ◽  
Monocytic Cells ◽  
Anionic Phospholipids ◽  
Phosphatidylserine Exposure ◽  
And Control

Oxidized cholesterol compounds or oxysterols are thought to be potent membrane-destabilizing agents. Anionic phospholipids, chiefly phosphatidylserine, have a procoagulant potential due to their ability to favour the membrane assembly of the characteristic clotting enzyme complexes including the tissue factor-dependent initiating complex. However, in resting cells, phosphatidylserine is sequestered in the inner leaflet of the plasma membrane. When THP-1 monocytic cells were cultured in the presence of 7β-hydroxycholesterol (7β-OH) or 25-hydroxycholesterol (25-OH), prothrombinase, which reflects anionic phospholipid exposure and tissue factor (TF) procoagulant activities, increased in a time- and dose-dependent manner. 7β-OH appeared 1.5- to 2-fold more potent than 25-OH. Interestingly, no effect of cholesterol itself could be detected on procoagulant activities. Nevertheless, no difference in TF activity could be detected between oxysterol-treated and control cells after disruption. TF antigen expression was the same in oxysterol-treated and control cells as shown by flow cytometry. In contrast, the use of labelled annexin V, a protein probe of anionic phospholipids, revealed an elevated number of cells with exposed phosphatidylserine. Because the latter also constitutes a signal for phagocyte recognition of apoptotic cells and fragments, and a proportion of cells displayed altered morphology with condensed chromatin and membrane blebs, analysis of DNA was performed and indicated apoptosis in oxysterol-treated cells. Hence, oxysterol-induced phosphatidylserine exposure and enhanced TF activity may result from apoptosis. These results suggest relationships between oxysterol and the amplification of coagulation reactions by monocytic cells resulting from induced phosphatidylserine exposure.

scholarly journals Micro Ribonucleic Acid Profiles of Traumatic Brain Injury Isolated From Plasma Extracellular Vesicles

Neurosurgery ◽  
2019 ◽  
Vol 66 (Supplement_1) ◽  
Author(s):  
Ross Puffer ◽  
Luz Cumba-Garcia ◽  
Benjamin T Himes ◽  
David O Okonkwo ◽  
Ian F Parney
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Extracellular Vesicles ◽  
Rna Isolation ◽  
Extracellular Vesicle ◽  
Micro Rna ◽  
Altered Consciousness ◽  
Rna Sequences ◽  
Altered Level ◽  
Level Of Consciousness

Abstract INTRODUCTION Extracellular vesicles (EVs) are membrane-bound particles released by the majority of human cells, including cells within the central nervous system. They may represent a diagnostic or prognostic target obtainable in peripheral blood of neurotrauma patients. We have isolated micro RNA sequences contained within EVs of 15 patients with traumatic brain injury (TBI) and compared them to miRNA sequences from 5 healthy controls. METHODS Extracellular vesicles were isolated from 15 TBI subjects, including 6 mild TBI (Glasgow Coma Scale (GCS) 13-15), 3 moderate TBI (GCS 9-12), and 6 severe TBI (GCS 3-8), as well as 5 healthy control. EVs were analyzed using nanoparticle tracking analysis. Samples underwent RNA isolation and extraction, followed by miRNA sequencing and analysis. RESULTS TBI patients presenting with an altered level of consciousness (GCS = 14) had a significantly higher mean extracellular vesicle size compared to subjects with normal GCS (mean + /− sem = 108.3 nm + /− 7.7 nm vs 89.2 nm + /− 6.7 nm; P < .04). GFAP ELISA of the samples demonstrated significantly higher GFAP concentration in subjects with altered level of consciousness (GCS = 14) as compared to those with normal GCS (mean + /− sem GFAP concentration 2204.2 pg/mL + /− 1067.2 pg/mL vs. 207.8 pg/mL + /− 270.8 pg/mL, P = .05). We identified 9 miRNA sequences that were found in a significantly higher proportion in patients with altered consciousness compared to controls, as well as 2 miRNA sequences that were significantly downregulated in subjects with altered consciousness as compared to controls. CONCLUSION EVs may contain brain specific biomarkers that are released in greater quantities after TBI. These molecules can be isolated from plasma and sequenced. Further analysis will better elucidate the final pathways affected by these up and downregulated miRNA sequences. Analysis of EVs in subjects with TBI may allow for the identification of novel diagnostic and potentially prognostic biomarkers.

scholarly journals Implantation of Neuronal Stem Cells Enhances Object Recognition without Increasing Neurogenesis after Lateral Fluid Percussion Injury in Mice

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Laura B. Ngwenya ◽  
Sarmistha Mazumder ◽  
Zachary R. Porter ◽  
Amy Minnema ◽  
Duane J. Oswald ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Stem Cell ◽  
Object Recognition ◽  
Brain Injury ◽  
Cell Transplantation ◽  
Hippocampal Neurogenesis ◽  
Cognitive Recovery ◽  
Fluid Percussion Injury ◽  
Immature Neurons ◽  
Lateral Fluid Percussion

Cognitive deficits after traumatic brain injury (TBI) are debilitating and contribute to the morbidity and loss of productivity of over 10 million people worldwide. Cell transplantation has been linked to enhanced cognitive function after experimental traumatic brain injury, yet the mechanism of recovery is poorly understood. Since the hippocampus is a critical structure for learning and memory, supports adult neurogenesis, and is particularly vulnerable after TBI, we hypothesized that stem cell transplantation after TBI enhances cognitive recovery by modulation of endogenous hippocampal neurogenesis. We performed lateral fluid percussion injury (LFPI) in adult mice and transplanted embryonic stem cell-derived neural progenitor cells (NPC). Our data confirm an injury-induced cognitive deficit in novel object recognition, a hippocampal-dependent learning task, which is reversed one week after NPC transplantation. While LFPI alone promotes hippocampal neurogenesis, as revealed by doublecortin immunolabeling of immature neurons, subsequent NPC transplantation prevents increased neurogenesis and is not associated with morphological maturation of endogenous injury-induced immature neurons. Thus, NPC transplantation enhances cognitive recovery early after LFPI without a concomitant increase in neuron numbers or maturation.

HOW DOES ANESTHESIA AFFECT VARIOUS LEVELS OF EXPERIMENTAL TRAUMATIC BRAIN INJURY?

2011 ◽  
Vol 04 (04) ◽  
pp. 409-420 ◽  
Author(s):  
BARBIRO-MICHAELY EFRAT ◽  
MANOR TAMAR ◽  
ROGATSKY GENNADY ◽  
MAYEVSKY AVRAHAM
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Severe Injuries ◽  
Doppler Flowmetry ◽  
Fluid Percussion Injury ◽  
Physiological Properties ◽  
Mitochondrial Redox State ◽  
Severe Tbi ◽  
Four Levels ◽  
Injured Patients

The use of anesthetics is a well-known treatment for severely injured patients. In the present study we tested the pathophysiology of several levels of injury damage in a rat model and also tested the effect of Equithesin on brain vitality in these models. Traumatic Brain Injury (TBI) was induced using the fluid percussion injury model in four levels: mild, moderate and two levels of severe TBI. Brain real-time evaluation was performed by the multiparametric monitoring assembly (MPA) which enable cerebral blood flow (CBF) monitoring by laser Doppler flowmetry, mitochondrial NADH (Nicotinamide adenine dinucleotide) monitoring by the fluorometric technique, ionic homehostasis using special mini-electrodes, intracranial pressure (ICP) by the ICP camino device and needle electrodes for ECoG (Electrocorticogram) recording. Our results showed high correlation between the level of impact and the extent of changes in the physiological properties of the injury as indicated by the changes in all parameters monitored using the MPA device. Moreover, Equithesin improved CBF, ionic extracellular level and mitochondrial redox state following mild and moderate TBI while in severe TBI, Equithesin did not improve the metabolic state of the cerebral cortex, although it decreased the mortality rate from 66% to 20%, and following extra-severe TBI level, Equithesin did not improve survival rate. In conclusion it seems that Equithesin's protective effect exists under mild to moderate levels of injury and not in case of severe injuries.

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