High-Resolution Confocal Fluorescence Imaging of Serine Hydrolase Activity in Cryosections - Application to Glioma Brain Unveils Activity Hotspots Originating from Tumor-Associated Neutrophils

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dc.contributor.author Aaltonen, Niina
dc.contributor.author Singha, Prosanta K.
dc.contributor.author Jakupovic, Hermina
dc.contributor.author Wirth, Thomas
dc.contributor.author Samaranayake, Haritha
dc.contributor.author Pasonen-Seppanen, Sanna
dc.contributor.author Rilla, Kirsi
dc.contributor.author Varjosalo, Markku
dc.contributor.author Edgington-Mitchell, Laura E.
dc.contributor.author Kasperkiewicz, Paulina
dc.contributor.author Drag, Marcin
dc.contributor.author Kälvälä, Sara
dc.contributor.author Moisio, Eemeli
dc.contributor.author Savinainen, Juha R.
dc.contributor.author Laitinen, Jarmo T.
dc.date.accessioned 2020-07-07T08:42:02Z
dc.date.available 2020-07-07T08:42:02Z
dc.date.issued 2020-03-15
dc.identifier.citation Aaltonen , N , Singha , P K , Jakupovic , H , Wirth , T , Samaranayake , H , Pasonen-Seppanen , S , Rilla , K , Varjosalo , M , Edgington-Mitchell , L E , Kasperkiewicz , P , Drag , M , Kälvälä , S , Moisio , E , Savinainen , J R & Laitinen , J T 2020 , ' High-Resolution Confocal Fluorescence Imaging of Serine Hydrolase Activity in Cryosections - Application to Glioma Brain Unveils Activity Hotspots Originating from Tumor-Associated Neutrophils ' , Biological Procedures Online , vol. 22 , no. 1 , 6 . https://doi.org/10.1186/s12575-020-00118-4
dc.identifier.other PURE: 140212825
dc.identifier.other PURE UUID: c0c698a8-9bf4-44a0-bc12-2ed174a583e2
dc.identifier.other WOS: 000520970600001
dc.identifier.other ORCID: /0000-0002-1340-9732/work/77087683
dc.identifier.uri http://hdl.handle.net/10138/317397
dc.description.abstract Background Serine hydrolases (SHs) are a functionally diverse family of enzymes playing pivotal roles in health and disease and have emerged as important therapeutic targets in many clinical conditions. Activity-based protein profiling (ABPP) using fluorophosphonate (FP) probes has been a powerful chemoproteomic approach in studies unveiling roles of SHs in various biological systems. ABPP utilizes cell/tissue proteomes and features the FP-warhead, linked to a fluorescent reporter for in-gel fluorescence imaging or a biotin tag for streptavidin enrichment and LC-MS/MS-based target identification. Existing ABPP approaches characterize global SH activity based on mobility in gel or MS-based target identification and cannot reveal the identity of the cell-type responsible for an individual SH activity originating from complex proteomes. Results Here, by using an activity probe with broad reactivity towards the SH family, we advance the ABPP methodology to glioma brain cryosections, enabling for the first time high-resolution confocal fluorescence imaging of global SH activity in the tumor microenvironment. Tumor-associated cell types were identified by extensive immunohistochemistry on activity probe-labeled sections. Tissue-ABPP indicated heightened SH activity in glioma vs. normal brain and unveiled activity hotspots originating from tumor-associated neutrophils (TANs), rather than tumor-associated macrophages (TAMs). Thorough optimization and validation was provided by parallel gel-based ABPP combined with LC-MS/MS-based target verification. Conclusions Our study advances the ABPP methodology to tissue sections, enabling high-resolution confocal fluorescence imaging of global SH activity in anatomically preserved complex native cellular environment. To achieve global portrait of SH activity throughout the section, a probe with broad reactivity towards the SH family members was employed. As ABPP requires no a priori knowledge of the identity of the target, we envisage no imaginable reason why the presently described approach would not work for sections regardless of species and tissue source. en
dc.format.extent 21
dc.language.iso eng
dc.relation.ispartof Biological Procedures Online
dc.rights cc_by
dc.rights.uri info:eu-repo/semantics/openAccess
dc.subject Activity-based protein profiling (ABPP)
dc.subject Brain cryosection
dc.subject Glioblastoma multiforme (GBM)
dc.subject Immunohistochemistry
dc.subject Serine hydrolase activity
dc.subject Neutrophil serine protease (NSP)
dc.subject TAMRA-FP probe
dc.subject Tumor-associated neutrophils
dc.subject INHIBITORS
dc.subject DISCOVERY
dc.subject PROTEASES
dc.subject AUTORADIOGRAPHY
dc.subject METABOLISM
dc.subject PROTEOMICS
dc.subject LANDSCAPE
dc.subject PROBES
dc.subject CELLS
dc.subject ABHD6
dc.subject 1182 Biochemistry, cell and molecular biology
dc.title High-Resolution Confocal Fluorescence Imaging of Serine Hydrolase Activity in Cryosections - Application to Glioma Brain Unveils Activity Hotspots Originating from Tumor-Associated Neutrophils en
dc.type Article
dc.contributor.organization University Management
dc.contributor.organization Institute of Biotechnology
dc.contributor.organization Helsinki Institute of Life Science HiLIFE, Joint Activities
dc.contributor.organization Molecular Systems Biology
dc.description.reviewstatus Peer reviewed
dc.relation.doi https://doi.org/10.1186/s12575-020-00118-4
dc.relation.issn 1480-9222
dc.rights.accesslevel openAccess
dc.type.version publishedVersion

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