Inhibition of NMDA receptor function with an anti-GluN1-S2 antibody impairs human platelet function and thrombosis
GluN1 is a mandatory component of N-methyl-D-aspartate receptors (NMDARs) best known for their roles in the brain, but with increasing evidence for relevance in peripheral tissues, including platelets. Certain anti-GluN1 antibodies reduce brain infarcts in rodent models of ischaemic stroke. There is...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2017-11-01
|
| Series: | Platelets |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/09537104.2017.1280149 |
| _version_ | 1827816995268067328 |
|---|---|
| author | Taryn N. Green Justin R. Hamilton Marie-Christine Morel-Kopp Zhaohua Zheng Ting-Yu T. Chen James I. Hearn Peng P. Sun Jack U. Flanagan Deborah Young P. Alan Barber Matthew J. During Christopher M. Ward Maggie L. Kalev-Zylinska |
| author_facet | Taryn N. Green Justin R. Hamilton Marie-Christine Morel-Kopp Zhaohua Zheng Ting-Yu T. Chen James I. Hearn Peng P. Sun Jack U. Flanagan Deborah Young P. Alan Barber Matthew J. During Christopher M. Ward Maggie L. Kalev-Zylinska |
| author_sort | Taryn N. Green |
| collection | DOAJ |
| description | GluN1 is a mandatory component of N-methyl-D-aspartate receptors (NMDARs) best known for their roles in the brain, but with increasing evidence for relevance in peripheral tissues, including platelets. Certain anti-GluN1 antibodies reduce brain infarcts in rodent models of ischaemic stroke. There is also evidence that human anti-GluN1 autoantibodies reduce neuronal damage in stroke patients, but the underlying mechanism is unclear. This study investigated whether anti-GluN1-mediated neuroprotection involves inhibition of platelet function. Four commercial anti-GluN1 antibodies were screened for their abilities to inhibit human platelet aggregation. Haematological parameters were examined in rats vaccinated with GluN1. Platelet effects of a mouse monoclonal antibody targeting the glycine-binding region of GluN1 (GluN1-S2) were tested in assays of platelet activation, aggregation and thrombus formation. The epitope of anti-GluN1-S2 was mapped and the mechanism of antibody action modelled using crystal structures of GluN1. Our work found that rats vaccinated with GluN1 had a mildly prolonged bleeding time and carried antibodies targeting mostly GluN1-S2. The monoclonal anti-GluN1-S2 antibody (from BD Biosciences) inhibited activation and aggregation of human platelets in the presence of adrenaline, adenosine diphosphate, collagen, thrombin and a protease-activated receptor 1-activating peptide. When human blood was flowed over collagen-coated surfaces, anti-GluN1-S2 impaired thrombus growth and stability. The epitope of anti-GluN1-S2 was mapped to α-helix H located within the glycine-binding clamshell of GluN1, where the antibody binding was computationally predicted to impair opening of the NMDAR channel. Our results indicate that anti-GluN1-S2 inhibits function of human platelets, including dense granule release and thrombus growth. Findings add to the evidence that platelet NMDARs regulate thrombus formation and suggest a novel mechanism by which anti-GluN1 autoantibodies limit stroke-induced neuronal damage. |
| first_indexed | 2024-03-12T00:28:12Z |
| format | Article |
| id | doaj.art-777a1eb7851b450190f5c33b151b9622 |
| institution | Directory Open Access Journal |
| issn | 0953-7104 1369-1635 |
| language | English |
| last_indexed | 2024-03-12T00:28:12Z |
| publishDate | 2017-11-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Platelets |
| spelling | doaj.art-777a1eb7851b450190f5c33b151b96222023-09-15T10:31:57ZengTaylor & Francis GroupPlatelets0953-71041369-16352017-11-0128879981110.1080/09537104.2017.12801491280149Inhibition of NMDA receptor function with an anti-GluN1-S2 antibody impairs human platelet function and thrombosisTaryn N. Green0Justin R. Hamilton1Marie-Christine Morel-Kopp2Zhaohua Zheng3Ting-Yu T. Chen4James I. Hearn5Peng P. Sun6Jack U. Flanagan7Deborah Young8P. Alan Barber9Matthew J. During10Christopher M. Ward11Maggie L. Kalev-Zylinska12University of AucklandAustralian Centre for Blood Diseases, Monash UniversityRoyal North Shore HospitalAustralian Centre for Blood Diseases, Monash UniversityUniversity of AucklandUniversity of AucklandUniversity of AucklandAuckland Cancer Society Research Centre, University of AucklandUniversity of AucklandAuckland City HospitalUniversity of AucklandRoyal North Shore HospitalUniversity of AucklandGluN1 is a mandatory component of N-methyl-D-aspartate receptors (NMDARs) best known for their roles in the brain, but with increasing evidence for relevance in peripheral tissues, including platelets. Certain anti-GluN1 antibodies reduce brain infarcts in rodent models of ischaemic stroke. There is also evidence that human anti-GluN1 autoantibodies reduce neuronal damage in stroke patients, but the underlying mechanism is unclear. This study investigated whether anti-GluN1-mediated neuroprotection involves inhibition of platelet function. Four commercial anti-GluN1 antibodies were screened for their abilities to inhibit human platelet aggregation. Haematological parameters were examined in rats vaccinated with GluN1. Platelet effects of a mouse monoclonal antibody targeting the glycine-binding region of GluN1 (GluN1-S2) were tested in assays of platelet activation, aggregation and thrombus formation. The epitope of anti-GluN1-S2 was mapped and the mechanism of antibody action modelled using crystal structures of GluN1. Our work found that rats vaccinated with GluN1 had a mildly prolonged bleeding time and carried antibodies targeting mostly GluN1-S2. The monoclonal anti-GluN1-S2 antibody (from BD Biosciences) inhibited activation and aggregation of human platelets in the presence of adrenaline, adenosine diphosphate, collagen, thrombin and a protease-activated receptor 1-activating peptide. When human blood was flowed over collagen-coated surfaces, anti-GluN1-S2 impaired thrombus growth and stability. The epitope of anti-GluN1-S2 was mapped to α-helix H located within the glycine-binding clamshell of GluN1, where the antibody binding was computationally predicted to impair opening of the NMDAR channel. Our results indicate that anti-GluN1-S2 inhibits function of human platelets, including dense granule release and thrombus growth. Findings add to the evidence that platelet NMDARs regulate thrombus formation and suggest a novel mechanism by which anti-GluN1 autoantibodies limit stroke-induced neuronal damage.http://dx.doi.org/10.1080/09537104.2017.1280149calciumglutamateion channelplatelet inhibitorstroke |
| spellingShingle | Taryn N. Green Justin R. Hamilton Marie-Christine Morel-Kopp Zhaohua Zheng Ting-Yu T. Chen James I. Hearn Peng P. Sun Jack U. Flanagan Deborah Young P. Alan Barber Matthew J. During Christopher M. Ward Maggie L. Kalev-Zylinska Inhibition of NMDA receptor function with an anti-GluN1-S2 antibody impairs human platelet function and thrombosis Platelets calcium glutamate ion channel platelet inhibitor stroke |
| title | Inhibition of NMDA receptor function with an anti-GluN1-S2 antibody impairs human platelet function and thrombosis |
| title_full | Inhibition of NMDA receptor function with an anti-GluN1-S2 antibody impairs human platelet function and thrombosis |
| title_fullStr | Inhibition of NMDA receptor function with an anti-GluN1-S2 antibody impairs human platelet function and thrombosis |
| title_full_unstemmed | Inhibition of NMDA receptor function with an anti-GluN1-S2 antibody impairs human platelet function and thrombosis |
| title_short | Inhibition of NMDA receptor function with an anti-GluN1-S2 antibody impairs human platelet function and thrombosis |
| title_sort | inhibition of nmda receptor function with an anti glun1 s2 antibody impairs human platelet function and thrombosis |
| topic | calcium glutamate ion channel platelet inhibitor stroke |
| url | http://dx.doi.org/10.1080/09537104.2017.1280149 |
| work_keys_str_mv | AT tarynngreen inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT justinrhamilton inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT mariechristinemorelkopp inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT zhaohuazheng inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT tingyutchen inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT jamesihearn inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT pengpsun inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT jackuflanagan inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT deborahyoung inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT palanbarber inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT matthewjduring inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT christophermward inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis AT maggielkalevzylinska inhibitionofnmdareceptorfunctionwithanantiglun1s2antibodyimpairshumanplateletfunctionandthrombosis |