ADAMTS-7 Modulates Atherosclerotic Plaque Formation by Degradation of TIMP-1

Circ Res. 2023 Sep 29;133(8):674-686. doi: 10.1161/CIRCRESAHA.123.322737. Epub 2023 Sep 7.

Authors

M Amin Sharifi^{1

2}, Michael Wierer³, Tan An Dang^{1

2}, Jelena Milic⁴, Aldo Moggio¹, Nadja Sachs⁵, Moritz von Scheidt^{1

2}, Julia Hinterdobler^{1

2}, Philipp Müller^{1

2}, Julia Werner^{1

2}, Barbara Stiller¹, Zouhair Aherrahrou^{6

7}, Jeanette Erdmann^{6

7}, Andrea Zaliani^{8

9}, Mira Graettinger^{8

9}, Jeanette Reinshagen^{8

9}, Sheraz Gul^{8

9}, Philip Gribbon^{8

9}, Lars Maegdefessel^{2

5}, Jürgen Bernhagen^{2

4}, Hendrik B Sager^{1

2}, Matthias Mann³, Heribert Schunkert^{1

2}, Thorsten Kessler^{1

2}

Affiliations

¹ Department of Cardiology, German Heart Centre Munich, Technical University of Munich, Germany (M.A.S., T.A.D., A.M., M.v.S., J.H., P.M., J.W., B.S., H.B.S., H.S., T.K.).
² German Centre for Cardiovascular Research (DZHK e.V.), partner site Munich Heart Alliance, Germany (M.A.S., T.A.D., M.v.S., J.H., P.M., J.W., L.M., J.B., H.B.S., H.S., T.K.).
³ Department of Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry, Martinsried, Germany (M.W., M.M.).
⁴ Division of Vascular Biology, Institute for Stroke and Dementia Research, Ludwig Maximilian University of Munich, Germany (J.M., J.B.).
⁵ Vascular Biology and Experimental Vascular Medicine Unit, Department of Vascular and Endovascular Surgery, Klinikum rechts der Isar, Technical University Munich, Germany (N.S., L.M.).
⁶ Institute for Cardiogenetics and University Heart Centre Lübeck, University of Lübeck, Germany (Z.A., J.E.).
⁷ German Centre for Cardiovascular Research (DZHK e.V.), partner site Hamburg/Kiel/Lübeck, Germany (Z.A., J.E.).
⁸ Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Hamburg, Germany (A.Z., M.G., J.R., S.G., P.G.).
⁹ Fraunhofer Cluster of Excellence for Immune-Mediated Diseases (CIMD), Hamburg, Germany (A.Z., M.G., J.R., S.G., P.G.).

Abstract

Background: The ADAMTS7 locus was genome-wide significantly associated with coronary artery disease. Lack of the ECM (extracellular matrix) protease ADAMTS-7 (A disintegrin and metalloproteinase-7) was shown to reduce atherosclerotic plaque formation. Here, we sought to identify molecular mechanisms and downstream targets of ADAMTS-7 mediating the risk of atherosclerosis.

Methods: Targets of ADAMTS-7 were identified by high-resolution mass spectrometry of atherosclerotic plaques from Apoe^-/- and Apoe^-/-Adamts7^-/- mice. ECM proteins were identified using solubility profiling. Putative targets were validated using immunofluorescence, in vitro degradation assays, coimmunoprecipitation, and Förster resonance energy transfer-based protein-protein interaction assays. ADAMTS7 expression was measured in fibrous caps of human carotid artery plaques.

Results: In humans, ADAMTS7 expression was higher in caps of unstable as compared to stable carotid plaques. Compared to Apoe^-/- mice, atherosclerotic aortas of Apoe^-/- mice lacking Adamts-7 (Apoe^-/-Adamts7^-/-) contained higher protein levels of Timp-1 (tissue inhibitor of metalloprotease-1). In coimmunoprecipitation experiments, the catalytic domain of ADAMTS-7 bound to TIMP-1, which was degraded in the presence of ADAMTS-7 in vitro. ADAMTS-7 reduced the inhibitory capacity of TIMP-1 at its canonical target MMP-9 (matrix metalloprotease-9). As a downstream mechanism, we investigated collagen content in plaques of Apoe^-/- and Apoe^-/-Adamts7^-/- mice after a Western diet. Picrosirius red staining of the aortic root revealed less collagen as a readout of higher MMP-9 activity in Apoe^-/- as compared to Apoe^-/- Adamts7^-/- mice. To facilitate high-throughput screening for ADAMTS-7 inhibitors with the aim of decreasing TIMP-1 degradation, we designed a Förster resonance energy transfer-based assay targeting the ADAMTS-7 catalytic site.

Conclusions: ADAMTS-7, which is induced in unstable atherosclerotic plaques, decreases TIMP-1 stability reducing its inhibitory effect on MMP-9, which is known to promote collagen degradation and is likewise associated with coronary artery disease. Disrupting the interaction of ADAMTS-7 and TIMP-1 might be a strategy to increase collagen content and plaque stability for the reduction of atherosclerosis-related events.

Keywords: aorta; atherosclerosis; disintegrins; mice; solubility.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ADAMTS7 Protein* / genetics
Animals
Atherosclerosis* / genetics
Collagen / metabolism
Coronary Artery Disease* / genetics
Humans
Matrix Metalloproteinase 9
Mice
Mice, Knockout, ApoE
Plaque, Atherosclerotic* / metabolism
Tissue Inhibitor of Metalloproteinase-1* / genetics
Tissue Inhibitor of Metalloproteinase-1* / metabolism

Substances

ADAMTS7 Protein
Collagen
Matrix Metalloproteinase 9
Tissue Inhibitor of Metalloproteinase-1
ADAMTS7 protein, human
Adamts7 protein, mouse

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