000157828 001__ 157828 000157828 005__ 20230915092359.0 000157828 0247_ $$2doi$$a10.1002/glia.23953 000157828 0247_ $$2pmid$$apmid:33314333 000157828 0247_ $$2ISSN$$a0894-1491 000157828 0247_ $$2ISSN$$a1098-1136 000157828 0247_ $$2altmetric$$aaltmetric:96174707 000157828 037__ $$aDZNE-2021-01285 000157828 041__ $$aEnglish 000157828 082__ $$a610 000157828 1001_ $$0P:(DE-2719)2811620$$aIbach, Melanie$$b0$$udzne 000157828 245__ $$aA reporter cell system for the triggering receptor expressed on myeloid cells 2 reveals differential effects of disease-associated variants on receptor signaling and activation by antibodies against the stalk region. 000157828 260__ $$aBognor Regis [u.a.]$$bWiley-Liss$$c2021 000157828 3367_ $$2DRIVER$$aarticle 000157828 3367_ $$2DataCite$$aOutput Types/Journal article 000157828 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article$$bjournal$$mjournal$$s1683797742_30879 000157828 3367_ $$2BibTeX$$aARTICLE 000157828 3367_ $$2ORCID$$aJOURNAL_ARTICLE 000157828 3367_ $$00$$2EndNote$$aJournal Article 000157828 520__ $$aThe triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor expressed on myeloid-derived cell types. The extracellular immunoglobulin-like domain of TREM2 binds anionic ligands including Apolipoprotein E and Amyloid-β. The transmembrane domain interacts with its adaptor protein DAP12/TYROBP that is responsible for propagation of downstream signaling upon ligand interaction. Several sequence variants of TREM2 have been linked to different neurodegenerative diseases including Alzheimer's disease. Here, we generated HEK 293 Flp-In cell lines stably expressing human TREM2 and DAP12 using a bicistronic construct with a T2A linker sequence allowing initial expression of both proteins in stoichiometric amounts. Cell biological and biochemical analyses revealed transport of TREM2 to the cell surface, and canonical sequential proteolytic processing and shedding of TREM2 (sTREM2). The functionality of this cell system was demonstrated by detection of phosphorylated spleen tyrosine kinase (SYK) upon stimulation of TREM2 with the anionic membrane lipid phosphatidylserine or anti-TREM2 antibodies. Using this cell model, we demonstrated impaired signaling of disease associated TREM2 variants. We also identified a monoclonal antibody against the stalk region of TREM2 with agonistic activity. Activation of TREM2-DAP12 signaling with the monoclonal antibody and the partial loss of function of disease associated variants were recapitulated in induced pluripotent stem cell derived microglia. Thus, this reporter cell model represents a suitable experimental system to investigate signaling of TREM2 variants, and for the identification of ligands and compounds that modulate TREM2-DAP12 signaling. MAIN POINTS: Disease associated variants impair the signaling activity of TREM2 by distinct mechanisms. Targeting the stalk region of TREM2 with bivalent antibodies activates TREM2 signaling. 000157828 536__ $$0G:(DE-HGF)POF4-352$$a352 - Disease Mechanisms (POF4-352)$$cPOF4-352$$fPOF IV$$x0 000157828 588__ $$aDataset connected to CrossRef, PubMed, , Journals: pub.dzne.de 000157828 650_7 $$2Other$$aTREM2 variants 000157828 650_7 $$2Other$$aagonistic antibody 000157828 650_7 $$2Other$$areporter system 000157828 650_7 $$2Other$$asignaling 000157828 650_2 $$2MeSH$$aAlzheimer Disease 000157828 650_2 $$2MeSH$$aAntibodies, Monoclonal 000157828 650_2 $$2MeSH$$aCarrier Proteins 000157828 650_2 $$2MeSH$$aHEK293 Cells 000157828 650_2 $$2MeSH$$aHumans 000157828 650_2 $$2MeSH$$aLigands 000157828 650_2 $$2MeSH$$aMembrane Glycoproteins: genetics 000157828 650_2 $$2MeSH$$aMicroglia 000157828 650_2 $$2MeSH$$aMyeloid Cells 000157828 650_2 $$2MeSH$$aReceptors, Immunologic: genetics 000157828 7001_ $$aMathews, Mona$$b1 000157828 7001_ $$aLinnartz-Gerlach, Bettina$$b2 000157828 7001_ $$aTheil, Sandra$$b3 000157828 7001_ $$aKumar, Sathish$$b4 000157828 7001_ $$0P:(DE-2719)2812867$$aFeederle, Regina$$b5$$udzne 000157828 7001_ $$0P:(DE-2719)9000037$$aBrüstle, Oliver$$b6$$udzne 000157828 7001_ $$00000-0002-5071-5202$$aNeumann, Harald$$b7 000157828 7001_ $$0P:(DE-2719)9000869$$aWalter, Jochen$$b8$$udzne 000157828 773__ $$0PERI:(DE-600)1474828-9$$a10.1002/glia.23953$$gVol. 69, no. 5, p. 1126 - 1139$$n5$$p1126 - 1139$$tGlia$$v69$$x1098-1136$$y2021 000157828 8564_ $$uhttps://pub.dzne.de/record/157828/files/DZNE-2021-01285.pdf$$yOpenAccess 000157828 8564_ $$uhttps://pub.dzne.de/record/157828/files/DZNE-2021-01285.pdf?subformat=pdfa$$xpdfa$$yOpenAccess 000157828 909CO $$ooai:pub.dzne.de:157828$$pdnbdelivery$$pdriver$$pVDB$$popen_access$$popenaire 000157828 9101_ $$0I:(DE-HGF)0$$6P:(DE-2719)2811620$$aExternal Institute$$b0$$kExtern 000157828 9101_ $$0I:(DE-588)1065079516$$6P:(DE-2719)2812867$$aDeutsches Zentrum für Neurodegenerative Erkrankungen$$b5$$kDZNE 000157828 9101_ $$0I:(DE-HGF)0$$6P:(DE-2719)9000037$$aExternal Institute$$b6$$kExtern 000157828 9131_ $$0G:(DE-HGF)POF4-352$$1G:(DE-HGF)POF4-350$$2G:(DE-HGF)POF4-300$$3G:(DE-HGF)POF4$$4G:(DE-HGF)POF$$aDE-HGF$$bGesundheit$$lNeurodegenerative Diseases$$vDisease Mechanisms$$x0 000157828 9141_ $$y2021 000157828 915__ $$0StatID:(DE-HGF)0160$$2StatID$$aDBCoverage$$bEssential Science Indicators$$d2021-01-29 000157828 915__ $$0StatID:(DE-HGF)1190$$2StatID$$aDBCoverage$$bBiological Abstracts$$d2021-01-29 000157828 915__ $$0LIC:(DE-HGF)CCBYNCND4$$2HGFVOC$$aCreative Commons Attribution-NonCommercial-NoDerivs CC BY-NC-ND 4.0 000157828 915__ $$0StatID:(DE-HGF)3001$$2StatID$$aDEAL Wiley$$d2021-01-29$$wger 000157828 915__ $$0StatID:(DE-HGF)0113$$2StatID$$aWoS$$bScience Citation Index Expanded$$d2021-01-29 000157828 915__ $$0StatID:(DE-HGF)0510$$2StatID$$aOpenAccess 000157828 915__ $$0StatID:(DE-HGF)0420$$2StatID$$aNationallizenz$$d2022-11-08$$wger 000157828 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS$$d2022-11-08 000157828 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline$$d2022-11-08 000157828 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bClarivate Analytics Master Journal List$$d2022-11-08 000157828 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection$$d2022-11-08 000157828 915__ $$0StatID:(DE-HGF)1050$$2StatID$$aDBCoverage$$bBIOSIS Previews$$d2022-11-08 000157828 915__ $$0StatID:(DE-HGF)1030$$2StatID$$aDBCoverage$$bCurrent Contents - Life Sciences$$d2022-11-08 000157828 915__ $$0StatID:(DE-HGF)0100$$2StatID$$aJCR$$bGLIA : 2021$$d2022-11-08 000157828 915__ $$0StatID:(DE-HGF)9905$$2StatID$$aIF >= 5$$bGLIA : 2021$$d2022-11-08 000157828 9201_ $$0I:(DE-2719)1140004$$kAG Feederle$$lAntibody production$$x0 000157828 9201_ $$0I:(DE-2719)1013004$$kAG Vorberg$$lPrion Cell Biology$$x1 000157828 980__ $$ajournal 000157828 980__ $$aVDB 000157828 980__ $$aUNRESTRICTED 000157828 980__ $$aI:(DE-2719)1140004 000157828 980__ $$aI:(DE-2719)1013004 000157828 9801_ $$aFullTexts