La Trobe
1161715_Siegel,P_2021.pdf (2.29 MB)

A DARPin targeting activated Mac-1 is a novel diagnostic tool and potential anti-inflammatory agent in myocarditis, sepsis and myocardial infarction

Download (2.29 MB)
journal contribution
posted on 10.05.2021, 00:23 by PM Siegel, I Bojti, N Bassler, J Holien, U Flierl, Xiaowei WangXiaowei Wang, P Waggershauser, X Tonnar, C Vedecnik, C Lamprecht, I Stankova, T Li, T Helbing, D Wolf, N Anto-Michel, LS Mitre, J Ehrlich, L Orlean, I Bender, A Przewosnik, M Mauler, L Hollederer, M Moser, C Bode, MW Parker, Karlheinz PeterKarlheinz Peter, P Diehl
The monocyte β -integrin Mac-1 is crucial for leukocyte–endothelium interaction, rendering it an attractive therapeutic target for acute and chronic inflammation. Using phage display, a Designed-Ankyrin-Repeat-Protein (DARPin) was selected as a novel binding protein targeting and blocking the α I-domain, an activation-specific epitope of Mac-1. This DARPin, named F7, specifically binds to activated Mac-1 on mouse and human monocytes as determined by flow cytometry. Homology modelling and docking studies defined distinct interaction sites which were verified by mutagenesis. Intravital microscopy showed reduced leukocyte–endothelium adhesion in mice treated with this DARPin. Using mouse models of sepsis, myocarditis and ischaemia/reperfusion injury, we demonstrate therapeutic anti-inflammatory effects. Finally, the activated Mac-1-specific DARPin is established as a tool to detect monocyte activation in patients receiving extra-corporeal membrane oxygenation, as well as suffering from sepsis and ST-elevation myocardial infarction. The activated Mac-1-specific DARPin F7 binds preferentially to activated monocytes, detects inflammation in critically ill patients, and inhibits monocyte and neutrophil function as an efficient new anti-inflammatory agent. 2 M

Funding

Patrick Siegel was supported by a Fellowship from the German Cardiological Society (DGK) and Philipp Diehl by a Fellowship from the German Research Foundation (DFG) and Monash University. Philipp Diehl and Istvan Bojti received a grant from the German Heart Research Foundation. Jessica Holien was supported by a joint Cure Cancer/Leukaemia Foundation Postdoctoral Fellowship. Karlheinz Peter was supported by the National Health and Medical Research Council of Australia (NHMRC) and the Australian Research Council (ARC). Michael Parker was supported by the National Health and Medical Research Council of Australia (NHMRC) and the Australian Cancer Research Foundation. Dennis Wolf received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (grant agreement No 853425) and from the "Else Kroner-Fresenius-Stiftung" (2019_A22).

History

Publication Date

15/03/2021

Journal

Basic Research in Cardiology

Volume

116

Issue

1

Article Number

17

Pagination

24p.

Publisher

Springer

ISSN

0300-8428

Rights Statement

The Author reserves all moral rights over the deposited text and must be credited if any re-use occurs. Documents deposited in OPAL are the Open Access versions of outputs published elsewhere. Changes resulting from the publishing process may therefore not be reflected in this document. The final published version may be obtained via the publisher’s DOI. Please note that additional copyright and access restrictions may apply to the published version.