Comparison of Anti-oncotic Effect of TRPM4 Blocking Antibody in Neuron, Astrocyte and Vascular Endothelial Cell Under Hypoxia

In stroke and other neurological diseases, Transient Receptor Potential Melastatin 4 (TRPM4) has been reported to cause oncotic cell death which is due to an excessive influx of sodium ions. Following stroke, hypoxia condition activates TRPM4 channel, and the sodium influx via TRPM4 is further enhan...

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Main Authors: Shunhui Wei, See Wee Low, Charlene Priscilla Poore, Bo Chen, Yahui Gao, Bernd Nilius, Ping Liao
Format: Article
Language:English
Published: Frontiers Media S.A. 2020-10-01
Series:Frontiers in Cell and Developmental Biology
Subjects:
Online Access:https://www.frontiersin.org/article/10.3389/fcell.2020.562584/full
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spelling doaj-4b214e4019784ec6963c5bf2b9a082512020-11-25T03:44:58ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2020-10-01810.3389/fcell.2020.562584562584Comparison of Anti-oncotic Effect of TRPM4 Blocking Antibody in Neuron, Astrocyte and Vascular Endothelial Cell Under HypoxiaShunhui Wei0See Wee Low1Charlene Priscilla Poore2Bo Chen3Yahui Gao4Bernd Nilius5Ping Liao6Ping Liao7Ping Liao8Calcium Signaling Laboratory, Department of Research, National Neuroscience Institute, Singapore, SingaporeCalcium Signaling Laboratory, Department of Research, National Neuroscience Institute, Singapore, SingaporeCalcium Signaling Laboratory, Department of Research, National Neuroscience Institute, Singapore, SingaporeCalcium Signaling Laboratory, Department of Research, National Neuroscience Institute, Singapore, SingaporeCalcium Signaling Laboratory, Department of Research, National Neuroscience Institute, Singapore, SingaporeDepartment of Cellular and Molecular Medicine, KU Leuven, Leuven, BelgiumCalcium Signaling Laboratory, Department of Research, National Neuroscience Institute, Singapore, SingaporeDuke-NUS Medical School, Singapore, SingaporeHealth and Social Sciences, Singapore Institute of Technology, Singapore, SingaporeIn stroke and other neurological diseases, Transient Receptor Potential Melastatin 4 (TRPM4) has been reported to cause oncotic cell death which is due to an excessive influx of sodium ions. Following stroke, hypoxia condition activates TRPM4 channel, and the sodium influx via TRPM4 is further enhanced by an increased TRPM4 expression. However, the effect of TRPM4 inhibition on oncotic cell death, particularly during the acute stage, remains largely unknown. Recently, we have developed a polyclonal antibody M4P that specifically inhibits TRPM4 channel. M4P blocks the channel via binding to a region close to the channel pore from extracellular space. Using M4P, we evaluated the acute effect of blocking TRPM4 in neurons, astrocytes, and vascular endothelial cells. In a rat stroke model, M4P co-localized with neuronal marker NeuN and endothelial marker vWF, whereas few GFAP positive astrocytes were stained by M4P in the ipsilateral hemisphere. When ATP was acutely depleted in cultured cortical neurons and microvascular endothelial cells, cell swelling was induced. Application of M4P significantly blocked TRPM4 current and attenuated oncosis. TUNEL assay, PI staining and western blot on cleaved Caspase-3 revealed that M4P could ameliorate apoptosis after 24 h hypoxia exposure. In contrast, acute ATP depletion in cultured astrocytes failed to demonstrate an increase of cell volume, and application of M4P or control IgG had no effect on cell volume change. When TRPM4 was overexpressed in astrocytes, acute ATP depletion successfully induced oncosis which could be suppressed by M4P treatment. Our results demonstrate that comparing to astrocytes, neurons, and vascular endothelial cells are more vulnerable to hypoxic injury. During the acute stage of stroke, blocking TRPM4 channel could protect neurons and vascular endothelial cells from oncotic cell death.https://www.frontiersin.org/article/10.3389/fcell.2020.562584/fullischemic strokeTRPM4swellingcell deaththerapeutic antibody
collection DOAJ
language English
format Article
sources DOAJ
author Shunhui Wei
See Wee Low
Charlene Priscilla Poore
Bo Chen
Yahui Gao
Bernd Nilius
Ping Liao
Ping Liao
Ping Liao
spellingShingle Shunhui Wei
See Wee Low
Charlene Priscilla Poore
Bo Chen
Yahui Gao
Bernd Nilius
Ping Liao
Ping Liao
Ping Liao
Comparison of Anti-oncotic Effect of TRPM4 Blocking Antibody in Neuron, Astrocyte and Vascular Endothelial Cell Under Hypoxia
Frontiers in Cell and Developmental Biology
ischemic stroke
TRPM4
swelling
cell death
therapeutic antibody
author_facet Shunhui Wei
See Wee Low
Charlene Priscilla Poore
Bo Chen
Yahui Gao
Bernd Nilius
Ping Liao
Ping Liao
Ping Liao
author_sort Shunhui Wei
title Comparison of Anti-oncotic Effect of TRPM4 Blocking Antibody in Neuron, Astrocyte and Vascular Endothelial Cell Under Hypoxia
title_short Comparison of Anti-oncotic Effect of TRPM4 Blocking Antibody in Neuron, Astrocyte and Vascular Endothelial Cell Under Hypoxia
title_full Comparison of Anti-oncotic Effect of TRPM4 Blocking Antibody in Neuron, Astrocyte and Vascular Endothelial Cell Under Hypoxia
title_fullStr Comparison of Anti-oncotic Effect of TRPM4 Blocking Antibody in Neuron, Astrocyte and Vascular Endothelial Cell Under Hypoxia
title_full_unstemmed Comparison of Anti-oncotic Effect of TRPM4 Blocking Antibody in Neuron, Astrocyte and Vascular Endothelial Cell Under Hypoxia
title_sort comparison of anti-oncotic effect of trpm4 blocking antibody in neuron, astrocyte and vascular endothelial cell under hypoxia
publisher Frontiers Media S.A.
series Frontiers in Cell and Developmental Biology
issn 2296-634X
publishDate 2020-10-01
description In stroke and other neurological diseases, Transient Receptor Potential Melastatin 4 (TRPM4) has been reported to cause oncotic cell death which is due to an excessive influx of sodium ions. Following stroke, hypoxia condition activates TRPM4 channel, and the sodium influx via TRPM4 is further enhanced by an increased TRPM4 expression. However, the effect of TRPM4 inhibition on oncotic cell death, particularly during the acute stage, remains largely unknown. Recently, we have developed a polyclonal antibody M4P that specifically inhibits TRPM4 channel. M4P blocks the channel via binding to a region close to the channel pore from extracellular space. Using M4P, we evaluated the acute effect of blocking TRPM4 in neurons, astrocytes, and vascular endothelial cells. In a rat stroke model, M4P co-localized with neuronal marker NeuN and endothelial marker vWF, whereas few GFAP positive astrocytes were stained by M4P in the ipsilateral hemisphere. When ATP was acutely depleted in cultured cortical neurons and microvascular endothelial cells, cell swelling was induced. Application of M4P significantly blocked TRPM4 current and attenuated oncosis. TUNEL assay, PI staining and western blot on cleaved Caspase-3 revealed that M4P could ameliorate apoptosis after 24 h hypoxia exposure. In contrast, acute ATP depletion in cultured astrocytes failed to demonstrate an increase of cell volume, and application of M4P or control IgG had no effect on cell volume change. When TRPM4 was overexpressed in astrocytes, acute ATP depletion successfully induced oncosis which could be suppressed by M4P treatment. Our results demonstrate that comparing to astrocytes, neurons, and vascular endothelial cells are more vulnerable to hypoxic injury. During the acute stage of stroke, blocking TRPM4 channel could protect neurons and vascular endothelial cells from oncotic cell death.
topic ischemic stroke
TRPM4
swelling
cell death
therapeutic antibody
url https://www.frontiersin.org/article/10.3389/fcell.2020.562584/full
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