Influence of Anticoagulants and Heparin Contaminants on the Suitability of MMP-9 as a Blood-Derived Biomarker

• Published in: International journal of molecular sciences Vol. 25; no. 18; p. 10106
• Main Authors: Küper, Daniela, Klos, Josefin, Kühl, Friederike, Attili, Rozan, Brand, Korbinian, Weissenborn, Karin, Lichtinghagen, Ralf, Huber, René
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 20.09.2024; MDPI
• Subjects: Anticoagulants; Anticoagulants (Medicine); Anticoagulants - pharmacology; Biological markers; Biomarkers; blood sampling; Chemokines; Chondroitin sulfate; Chondroitin Sulfates - pharmacology; contaminants; Contamination; Cytokines; Drug adulteration; Drug Contamination; Fondaparinux - pharmacology; Health aspects; Heparan sulfate; heparin; Heparin - pharmacology; Hirudins - pharmacology; Humans; Hyaluronic acid; Hyaluronic Acid - pharmacology; Jurkat Cells; Lymphocytes; Matrix Metalloproteinase 9 - genetics; Matrix Metalloproteinase 9 - metabolism; Medical research; Medicine, Experimental; Metalloenzymes; MMP-9; Monocytes - drug effects; Monocytes - metabolism; over-sulfated chondroitin sulfate; Physiological aspects; THP-1 Cells; Germany; blood sampling; contaminants; heparin; anticoagulants; over-sulfated chondroitin sulfate; MMP-9
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms251810106

In contrast to other common anticoagulants such as citrate and low-molecular-weight heparin (LMWH), high-molecular-weight heparin (HMWH) induces the expression of matrix metalloproteinase (MMP)-9, which is also measured as a biomarker for stroke in blood samples. Mechanistically, HMWH-stimulated T cells produce cytokines that induce monocytic MMP-9 expression. Here, the influence of further anticoagulants (Fondaparinux, Hirudin, and Alteplase) and the heparin-contaminating glycosaminoglycans (GAG) hyaluronic acid (HA), dermatan sulfate (DS), chondroitin sulfate (CS), and over-sulfated CS (OSCS) on MMP-9 was analyzed to assess its suitability as a biomarker under various conditions. Therefore, starved Jurkat T cells were stimulated with anticoagulants/contaminants. Subsequently, starved monocytic THP-1 cells were incubated with the conditioned Jurkat supernatant, and MMP-9 mRNA levels were monitored (quantitative (q)PCR). Jurkat-derived mediators secreted in response to anticoagulants/contaminants were also assessed (proteome profiler array). The supernatants of HMWH-, Hirudin-, CS-, and OSCS-treated Jurkat cells comprised combinations of activating mediators and led to a significant (in the case of OSCS, dramatic) MMP-9 induction in THP-1. HA induced MMP-9 only in high concentrations, while LMWH, Fondaparinux, Alteplase, and DS had no effect. This indicates that depending on molecular weight and charge (but independent of anticoagulant activity), anticoagulants/contaminants provoke the expression of T-cell-derived cytokines/chemokines that induce monocytic MMP-9 expression, thus potentially impairing the diagnostic validity of MMP-9.