Development and characterization of a monoclonal antibody blocking human TRPM4 channel

Abstract TRPM4 is a calcium-activated non-selective monovalent cation channel implicated in diseases such as stroke. Lack of potent and selective inhibitors remains a major challenge for studying TRPM4. Using a polypeptide from rat TRPM4, we have generated a polyclonal antibody M4P which could allev...

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Autores principales:	See Wee Low, Yahui Gao, Shunhui Wei, Bo Chen, Bernd Nilius, Ping Liao
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4e73928161fe44fea160522f5b96ca9b
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spelling	oai:doaj.org-article:4e73928161fe44fea160522f5b96ca9b2021-12-02T15:53:10ZDevelopment and characterization of a monoclonal antibody blocking human TRPM4 channel10.1038/s41598-021-89935-52045-2322https://doaj.org/article/4e73928161fe44fea160522f5b96ca9b2021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89935-5https://doaj.org/toc/2045-2322Abstract TRPM4 is a calcium-activated non-selective monovalent cation channel implicated in diseases such as stroke. Lack of potent and selective inhibitors remains a major challenge for studying TRPM4. Using a polypeptide from rat TRPM4, we have generated a polyclonal antibody M4P which could alleviate reperfusion injury in a rat model of stroke. Here, we aim to develop a monoclonal antibody that could block human TRPM4 channel. Two mouse monoclonal antibodies M4M and M4M1 were developed to target an extracellular epitope of human TRPM4. Immunohistochemistry and western blot were used to characterize the binding of these antibodies to human TRPM4. Potency of inhibition was compared using electrophysiological methods. We further evaluated the therapeutic potential on a rat model of middle cerebral artery occlusion. Both M4M and M4M1 could bind to human TRPM4 channel on the surface of live cells. Prolonged incubation with TRPM4 blocking antibody internalized surface TRPM4. Comparing to M4M1, M4M is more effective in blocking human TRPM4 channel. In human brain microvascular endothelial cells, M4M successfully inhibited TRPM4 current and ameliorated hypoxia-induced cell swelling. Using wild type rats, neither antibody demonstrated therapeutic potential on stroke. Human TRPM4 channel can be blocked by a monoclonal antibody M4M targeting a key antigenic sequence. For future clinical translation, the antibody needs to be humanized and a transgenic animal carrying human TRPM4 sequence is required for in vivo characterizing its therapeutic potential.See Wee LowYahui GaoShunhui WeiBo ChenBernd NiliusPing LiaoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q See Wee Low Yahui Gao Shunhui Wei Bo Chen Bernd Nilius Ping Liao Development and characterization of a monoclonal antibody blocking human TRPM4 channel
description	Abstract TRPM4 is a calcium-activated non-selective monovalent cation channel implicated in diseases such as stroke. Lack of potent and selective inhibitors remains a major challenge for studying TRPM4. Using a polypeptide from rat TRPM4, we have generated a polyclonal antibody M4P which could alleviate reperfusion injury in a rat model of stroke. Here, we aim to develop a monoclonal antibody that could block human TRPM4 channel. Two mouse monoclonal antibodies M4M and M4M1 were developed to target an extracellular epitope of human TRPM4. Immunohistochemistry and western blot were used to characterize the binding of these antibodies to human TRPM4. Potency of inhibition was compared using electrophysiological methods. We further evaluated the therapeutic potential on a rat model of middle cerebral artery occlusion. Both M4M and M4M1 could bind to human TRPM4 channel on the surface of live cells. Prolonged incubation with TRPM4 blocking antibody internalized surface TRPM4. Comparing to M4M1, M4M is more effective in blocking human TRPM4 channel. In human brain microvascular endothelial cells, M4M successfully inhibited TRPM4 current and ameliorated hypoxia-induced cell swelling. Using wild type rats, neither antibody demonstrated therapeutic potential on stroke. Human TRPM4 channel can be blocked by a monoclonal antibody M4M targeting a key antigenic sequence. For future clinical translation, the antibody needs to be humanized and a transgenic animal carrying human TRPM4 sequence is required for in vivo characterizing its therapeutic potential.
format	article
author	See Wee Low Yahui Gao Shunhui Wei Bo Chen Bernd Nilius Ping Liao
author_facet	See Wee Low Yahui Gao Shunhui Wei Bo Chen Bernd Nilius Ping Liao
author_sort	See Wee Low
title	Development and characterization of a monoclonal antibody blocking human TRPM4 channel
title_short	Development and characterization of a monoclonal antibody blocking human TRPM4 channel
title_full	Development and characterization of a monoclonal antibody blocking human TRPM4 channel
title_fullStr	Development and characterization of a monoclonal antibody blocking human TRPM4 channel
title_full_unstemmed	Development and characterization of a monoclonal antibody blocking human TRPM4 channel
title_sort	development and characterization of a monoclonal antibody blocking human trpm4 channel
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/4e73928161fe44fea160522f5b96ca9b
work_keys_str_mv	AT seeweelow developmentandcharacterizationofamonoclonalantibodyblockinghumantrpm4channel AT yahuigao developmentandcharacterizationofamonoclonalantibodyblockinghumantrpm4channel AT shunhuiwei developmentandcharacterizationofamonoclonalantibodyblockinghumantrpm4channel AT bochen developmentandcharacterizationofamonoclonalantibodyblockinghumantrpm4channel AT berndnilius developmentandcharacterizationofamonoclonalantibodyblockinghumantrpm4channel AT pingliao developmentandcharacterizationofamonoclonalantibodyblockinghumantrpm4channel
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Development and characterization of a monoclonal antibody blocking human TRPM4 channel

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