In vivo evaluation of monoclonal antibody M4M using a humanised rat model of stroke demonstrates attenuation of reperfusion injury via blocking human TRPM4 channel
BACKGROUND: Blocking Transient Receptor Potential Melastatin 4 (TRPM4) in rodents by our antibody M4P has shown to attenuate cerebral ischaemia-reperfusion injury. Since M4P does not interact with human TRPM4, the therapeutic potential of blocking human TRPM4 remains unclear. We developed a monoclonal antibody M4M that inhibited human TRPM4 in cultured cells. However, M4M has no effect on stroke outcome in wild-type rats. Therefore, M4M needs to be evaluated on animal models expressing human TRPM4.
METHODS: We generated a humanised rat model using the CRISPR/Cas technique to knock-in (KI) the human TRPM4 antigen sequence.
RESULTS: In primary neurons from human TRPM4 KI rats, M4M binds to hypoxic neurons, but not normoxic nor wild-type neurons. Electrophysiological studies showed that M4M blocked ATP depletion-induced activation of TRPM4 and inhibited hypoxia-associated cell volume increase. In a stroke model, administration of M4M reduced infarct volume in KI rats. Rotarod test and Neurological deficit score revealed improvement following M4M treatment.
CONCLUSION: M4M selectively binds and inhibits hypoxia-induced human TRPM4 channel activation in neurons from the humanised rat model, with no effect on healthy neurons. Use of M4M in stroke rats showed functional improvements, suggesting the potential for anti-human TRPM4 antibodies in treating acute ischaemic stroke patients.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:32 |
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| Enthalten in: |
Journal of drug targeting - 32(2024), 4 vom: 20. Apr., Seite 413-422 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Poore, Charlene Priscilla [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 03.04.2024 Date Revised 03.04.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1080/1061186X.2024.2313522 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM368351696 |
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| 100 | 1 | |a Poore, Charlene Priscilla |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a In vivo evaluation of monoclonal antibody M4M using a humanised rat model of stroke demonstrates attenuation of reperfusion injury via blocking human TRPM4 channel |
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| 500 | |a Date Revised 03.04.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a BACKGROUND: Blocking Transient Receptor Potential Melastatin 4 (TRPM4) in rodents by our antibody M4P has shown to attenuate cerebral ischaemia-reperfusion injury. Since M4P does not interact with human TRPM4, the therapeutic potential of blocking human TRPM4 remains unclear. We developed a monoclonal antibody M4M that inhibited human TRPM4 in cultured cells. However, M4M has no effect on stroke outcome in wild-type rats. Therefore, M4M needs to be evaluated on animal models expressing human TRPM4 | ||
| 520 | |a METHODS: We generated a humanised rat model using the CRISPR/Cas technique to knock-in (KI) the human TRPM4 antigen sequence | ||
| 520 | |a RESULTS: In primary neurons from human TRPM4 KI rats, M4M binds to hypoxic neurons, but not normoxic nor wild-type neurons. Electrophysiological studies showed that M4M blocked ATP depletion-induced activation of TRPM4 and inhibited hypoxia-associated cell volume increase. In a stroke model, administration of M4M reduced infarct volume in KI rats. Rotarod test and Neurological deficit score revealed improvement following M4M treatment | ||
| 520 | |a CONCLUSION: M4M selectively binds and inhibits hypoxia-induced human TRPM4 channel activation in neurons from the humanised rat model, with no effect on healthy neurons. Use of M4M in stroke rats showed functional improvements, suggesting the potential for anti-human TRPM4 antibodies in treating acute ischaemic stroke patients | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a TRPM4 channel | |
| 650 | 4 | |a acute ischaemic stroke | |
| 650 | 4 | |a humanised rat model | |
| 650 | 4 | |a hypoxia | |
| 650 | 4 | |a monoclonal antibody | |
| 650 | 4 | |a reperfusion injury | |
| 650 | 7 | |a Transient Receptor Potential Channels |2 NLM | |
| 650 | 7 | |a Antibodies, Monoclonal |2 NLM | |
| 650 | 7 | |a TRPM Cation Channels |2 NLM | |
| 650 | 7 | |a TRPM4 protein, human |2 NLM | |
| 650 | 7 | |a TRPM4 protein, rat |2 NLM | |
| 700 | 1 | |a Wei, Shunhui |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Bo |e verfasserin |4 aut | |
| 700 | 1 | |a Low, See Wee |e verfasserin |4 aut | |
| 700 | 1 | |a Tan, Jeslyn Si Qi |e verfasserin |4 aut | |
| 700 | 1 | |a Lee, Andy Thiam-Huat |e verfasserin |4 aut | |
| 700 | 1 | |a Nilius, Bernd |e verfasserin |4 aut | |
| 700 | 1 | |a Liao, Ping |e verfasserin |4 aut | |
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