Clin Res Cardiol (2023). https://doi.org/10.1007/s00392-023-02180-w

The novel Coronary Artery Disease Risk Factor Adamts-7 degrades TIMP-1 In Atherosclerosis

A. Sharifi1, M. Wierer2, T. A. Dang1, J. Milic3, A. Moggio1, J. Hinterdobler1, P. Müller1, J. Werner1, B. Stiller1, Z. Aherrahrou4, J. Erdmann5, S. Gul6, P. Gribbon6, L. Maegdefessel7, J. Bernhagen3, H. Sager1, M. Mann2, H. Schunkert1, T. Keßler1

1Klinik für Herz- und Kreislauferkrankungen, Deutsches Herzzentrum München, München; 2Max-Planck-Institut für Biochemie, Martinsried; 3Institut für Schlaganfall- und Demenzforschung, München; 4Institut für Kardiogenetik, Universität Lübeck, Lübeck; 5Institut für Kardiogenetik, Universitätsklinikum Schleswig-Holstein, Lübeck; 6Fraunhofer Institute for Translational Medicine and Pharmacology, Hamburg; 7Klinik für Vaskuläre und Endovaskuläre Chirurgie, Klinikum rechts der Isar der Technischen Universität München, München;

Background: The extracellular matrix (ECM) protease ADAMTS-7 was associated with coronary artery disease (CAD) in genome-wide association studies. ADAMTS-7 was shown to be expressed at all stages in human plaques. Mice lacking Adamts-7 displayed reduced atherosclerotic plaque formation. These findings render ADAMTS-7 a promising therapeutic target; the underlying molecular and cellular mechanisms remain, however, unknown. Methods and Results: Here, we aimed at identifying putative downstream targets of ADAMTS-7 in atherosclerotis. Using high-resolution mass spectrometry of atherosclerotic plaques in Apoe-/- and Apoe-/-Adamts7-/- mice, targets of Adamts-7 were identified. We specifically investigated ECM proteins using solubility profiling and identified the endogenous inhibitor of matrix metalloproteinases (MMP) Timp-1 as a novel target of Adamts-7. Adamts-7 and Timp-1 were co-localized in atherosclerotic plaques. Co-immunoprecipitation (Co-IP) experiments revealed binding of TIMP-1 to the catalytic domain of ADAMTS-7 and in vitro degradation assays demonstrated degradation of TIMP-1 by ADAMTS-7. Co-IP furthermore showed less binding of TIMP-1 to its canonical target MMP-9 in the presence of ADAMTS-7. This scaffolding and degradation of TIMP-1 by ADAMTS-7 subsequently impaired TIMP-1-mediated inhibition of MMP-9 in vitro. We next investigated collagen content in atherosclerotic plaques of Apoe-/- and Apoe-/- Adamts7-/- mice after Western diet. Picosirius red stainings of the aortic root revealed less collagen content as a readout of higher MMP-9 activity in Apoe-/- as compared to Apoe-/- Adamts7-/- mice. Since ADAMTS-7 might exert its pro-atherogenic effects through interaction with TIMP-1, we established a TIMP-1-ADAMTS-7-interaction assay based on Förster resonance energy transfer (FRET) to identify inhibitors of this protein-protein interaction. Conclusion and Outlook: TIMP-1 represents a novel target of ADAMTS-7. Its degradation can explain the role of this novel risk factor in CAD. The FRET-based protein-protein interaction assay can be used for high-throughput screening to identifiy inhibitors of atherosclerosic plaque formation and progression.

https://dgk.org/kongress_programme/jt2023/aV61.html