TREM2 shedding by cleavage at the H157‐S158 bond is accelerated for the Alzheimer's disease‐associated H157Y variant
Abstract We have characterised the proteolytic cleavage events responsible for the shedding of triggering receptor expressed on myeloid cells 2 (TREM2) from primary cultures of human macrophages, murine microglia and TREM2‐expressing human embryonic kidney (HEK293) cells. In all cell types, a solubl...
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Springer Nature
2017-08-01
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| Series: | EMBO Molecular Medicine |
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| Online Access: | https://doi.org/10.15252/emmm.201707673 |
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| author | Peter Thornton Jean Sevalle Michael J Deery Graham Fraser Ye Zhou Sara Ståhl Elske H Franssen Roger B Dodd Seema Qamar Beatriz Gomez Perez‐Nievas Louise SC Nicol Susanna Eketjäll Jefferson Revell Clare Jones Andrew Billinton Peter H St George‐Hyslop Iain Chessell Damian C Crowther |
| author_facet | Peter Thornton Jean Sevalle Michael J Deery Graham Fraser Ye Zhou Sara Ståhl Elske H Franssen Roger B Dodd Seema Qamar Beatriz Gomez Perez‐Nievas Louise SC Nicol Susanna Eketjäll Jefferson Revell Clare Jones Andrew Billinton Peter H St George‐Hyslop Iain Chessell Damian C Crowther |
| author_sort | Peter Thornton |
| collection | DOAJ |
| description | Abstract We have characterised the proteolytic cleavage events responsible for the shedding of triggering receptor expressed on myeloid cells 2 (TREM2) from primary cultures of human macrophages, murine microglia and TREM2‐expressing human embryonic kidney (HEK293) cells. In all cell types, a soluble 17 kDa N‐terminal cleavage fragment was shed into the conditioned media in a constitutive process that is inhibited by G1254023X and metalloprotease inhibitors and siRNA targeting ADAM10. Inhibitors of serine proteases and matrix metalloproteinases 2/9, and ADAM17 siRNA did not block TREM2 shedding. Peptidomimetic protease inhibitors highlighted a possible cleavage site, and mass spectrometry confirmed that shedding occurred predominantly at the H157‐S158 peptide bond for both wild‐type and H157Y human TREM2 and for the wild‐type murine orthologue. Crucially, we also show that the Alzheimer's disease‐associated H157Y TREM2 variant was shed more rapidly than wild type from HEK293 cells, possibly by a novel, batimastat‐ and ADAM10‐siRNA‐independent, sheddase activity. These insights offer new therapeutic targets for modulating the innate immune response in Alzheimer's and other neurological diseases. |
| format | Article |
| id | doaj-art-c069149e8d3140e48dd7f555f57c2d71 |
| institution | DOAJ |
| issn | 1757-4676 1757-4684 |
| language | English |
| publishDate | 2017-08-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | EMBO Molecular Medicine |
| spelling | doaj-art-c069149e8d3140e48dd7f555f57c2d712025-08-20T03:06:00ZengSpringer NatureEMBO Molecular Medicine1757-46761757-46842017-08-019101366137810.15252/emmm.201707673TREM2 shedding by cleavage at the H157‐S158 bond is accelerated for the Alzheimer's disease‐associated H157Y variantPeter Thornton0Jean Sevalle1Michael J Deery2Graham Fraser3Ye Zhou4Sara Ståhl5Elske H Franssen6Roger B Dodd7Seema Qamar8Beatriz Gomez Perez‐Nievas9Louise SC Nicol10Susanna Eketjäll11Jefferson Revell12Clare Jones13Andrew Billinton14Peter H St George‐Hyslop15Iain Chessell16Damian C Crowther17Neuroscience, Innovative Medicines and Early Development, AstraZeneca, Granta ParkTanz Centre for Research in Neurodegenerative Diseases, University of TorontoCambridge Centre for Proteomics, University of CambridgeNeuroscience, Innovative Medicines and Early Development, AstraZeneca, Granta ParkTanz Centre for Research in Neurodegenerative Diseases, University of TorontoAstraZeneca Translational Sciences Centre, Karolinska InstitutetNeuroscience, Innovative Medicines and Early Development, AstraZeneca, Granta ParkDepartment of Clinical Neurosciences, Cambridge Institute for Medical Research, University of CambridgeDepartment of Clinical Neurosciences, Cambridge Institute for Medical Research, University of CambridgeNeuroscience, Innovative Medicines and Early Development, AstraZeneca, Granta ParkMedImmune Limited, Granta ParkCardiovascular and Metabolic Diseases, Innovative Medicines and Early Development, AstraZeneca, ICMCMedImmune Limited, Granta ParkMedImmune Limited, Granta ParkNeuroscience, Innovative Medicines and Early Development, AstraZeneca, Granta ParkTanz Centre for Research in Neurodegenerative Diseases, University of TorontoNeuroscience, Innovative Medicines and Early Development, AstraZeneca, Granta ParkNeuroscience, Innovative Medicines and Early Development, AstraZeneca, Granta ParkAbstract We have characterised the proteolytic cleavage events responsible for the shedding of triggering receptor expressed on myeloid cells 2 (TREM2) from primary cultures of human macrophages, murine microglia and TREM2‐expressing human embryonic kidney (HEK293) cells. In all cell types, a soluble 17 kDa N‐terminal cleavage fragment was shed into the conditioned media in a constitutive process that is inhibited by G1254023X and metalloprotease inhibitors and siRNA targeting ADAM10. Inhibitors of serine proteases and matrix metalloproteinases 2/9, and ADAM17 siRNA did not block TREM2 shedding. Peptidomimetic protease inhibitors highlighted a possible cleavage site, and mass spectrometry confirmed that shedding occurred predominantly at the H157‐S158 peptide bond for both wild‐type and H157Y human TREM2 and for the wild‐type murine orthologue. Crucially, we also show that the Alzheimer's disease‐associated H157Y TREM2 variant was shed more rapidly than wild type from HEK293 cells, possibly by a novel, batimastat‐ and ADAM10‐siRNA‐independent, sheddase activity. These insights offer new therapeutic targets for modulating the innate immune response in Alzheimer's and other neurological diseases.https://doi.org/10.15252/emmm.201707673genetic riskmicroglianeurodegenerationneuroinflammation |
| spellingShingle | Peter Thornton Jean Sevalle Michael J Deery Graham Fraser Ye Zhou Sara Ståhl Elske H Franssen Roger B Dodd Seema Qamar Beatriz Gomez Perez‐Nievas Louise SC Nicol Susanna Eketjäll Jefferson Revell Clare Jones Andrew Billinton Peter H St George‐Hyslop Iain Chessell Damian C Crowther TREM2 shedding by cleavage at the H157‐S158 bond is accelerated for the Alzheimer's disease‐associated H157Y variant EMBO Molecular Medicine genetic risk microglia neurodegeneration neuroinflammation |
| title | TREM2 shedding by cleavage at the H157‐S158 bond is accelerated for the Alzheimer's disease‐associated H157Y variant |
| title_full | TREM2 shedding by cleavage at the H157‐S158 bond is accelerated for the Alzheimer's disease‐associated H157Y variant |
| title_fullStr | TREM2 shedding by cleavage at the H157‐S158 bond is accelerated for the Alzheimer's disease‐associated H157Y variant |
| title_full_unstemmed | TREM2 shedding by cleavage at the H157‐S158 bond is accelerated for the Alzheimer's disease‐associated H157Y variant |
| title_short | TREM2 shedding by cleavage at the H157‐S158 bond is accelerated for the Alzheimer's disease‐associated H157Y variant |
| title_sort | trem2 shedding by cleavage at the h157 s158 bond is accelerated for the alzheimer s disease associated h157y variant |
| topic | genetic risk microglia neurodegeneration neuroinflammation |
| url | https://doi.org/10.15252/emmm.201707673 |
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