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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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

Title: High-Resolution Confocal Fluorescence Imaging of Serine Hydrolase Activity in Cryosections - Application to Glioma Brain Unveils Activity Hotspots Originating from Tumor-Associated Neutrophils
Author: Aaltonen, Niina; Singha, Prosanta K.; Jakupovic, Hermina; Wirth, Thomas; Samaranayake, Haritha; Pasonen-Seppanen, Sanna; Rilla, Kirsi; Varjosalo, Markku; Edgington-Mitchell, Laura E.; Kasperkiewicz, Paulina; Drag, Marcin; Kälvälä, Sara; Moisio, Eemeli; Savinainen, Juha R.; Laitinen, Jarmo T.
Other contributor: University of Helsinki, University of Eastern Finland (UEF)
University of Helsinki, University Management



Date: 2020-03-15
Language: eng
Number of pages: 21
Belongs to series: Biological Procedures Online
ISSN: 1480-9222
DOI: https://doi.org/10.1186/s12575-020-00118-4
URI: http://hdl.handle.net/10138/317397
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.
Subject: Activity-based protein profiling (ABPP)
Brain cryosection
Glioblastoma multiforme (GBM)
Immunohistochemistry
Serine hydrolase activity
Neutrophil serine protease (NSP)
TAMRA-FP probe
Tumor-associated neutrophils
INHIBITORS
DISCOVERY
PROTEASES
AUTORADIOGRAPHY
METABOLISM
PROTEOMICS
LANDSCAPE
PROBES
CELLS
ABHD6
1182 Biochemistry, cell and molecular biology
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