Bivalent Inhibitor with Selectivity for Trimeric MMP-9 Amplifies Neutrophil Chemotaxis and Enables Functional Studies on MMP-9 Proteoforms

Abstract

<jats:p>A fundamental part of the immune response to infection or injury is leukocyte migration. Matrix metalloproteinases (MMPs) are a class of secreted or cell-bound endopeptidases, implicated in every step of the process of inflammatory cell migration. Hence, specific inhibition of MMPs is an interesting approach to control inflammation. We evaluated the potential of a bivalent carboxylate inhibitor to selectively inhibit the trimeric proteoform of MMP-9 and compared this with a corresponding monovalent inhibitor. The bivalent inhibitor efficiently inhibited trimeric MMP-9 (IC50 = 0.1 nM), with at least 500-fold selectivity for MMP-9 trimers over monomers. Surprisingly, in a mouse model for chemotaxis, the bivalent inhibitor amplified leukocyte influxes towards lipopolysaccharide-induced inflammation. We verified by microscopic and flow cytometry analysis increased amounts of neutrophils. In a mouse model for endotoxin shock, mice treated with the bivalent inhibitor had significantly increased levels of MMP-9 in plasma and lungs, indicative for increased inflammation. In conclusion, we propose a new role for MMP-9 trimers in tempering excessive neutrophil migration. In addition, we have identified a small molecule inhibitor with a high selectivity for the trimeric proteoform of MMP-9, which will allow further research on the functions of MMP-9 proteoforms.</jats:p>

Internal Disulfide Bonding and Glycosylation of Interleukin-7 Protect Against Proteolytic Inactivation by Neutrophil Metalloproteinases and Serine Proteases

“…Recombinant human MMP-9 was produced, purified, activated and tested as described ( 27 ). Recombinant human proMMP-2, proMMP-7, proMMP-8 and ‘a disintegrin and metalloproteases’ (ADAM)17 were purchased from R&D Systems and activated as previously described ( 28 ). Active human NE was purchased from Abcam (cat.…”

“…Neutrophil degranulates were collected from human blood granulocytes stimulated with N-formylmethionyl-leucyl-fenylalanine (fMLF), as previously described ( 21 , 28 ). All blood donors gave written informed consent in accordance with the Declaration of Helsinki.…”

Hypoxia- and Inflammation-Related Transcription Factor SP3 May Be Involved in Platelet Activation and Inflammation in Intracranial Hemorrhage

“…TIMP1 and PLAUR are involved in inflammation and hypoxia-related progression in ICH. MMP-9 is expressed in inflamed tissues and is involved in the inflammatory process ( 61 ). TIMP-1 is a natural inhibitor of MMP-9 ( 62 ).…”

Identification of differentially expressed genes, transcription factors, microRNAs and pathways in neutrophils of sepsis patients through bioinformatics analysis

“…Meanwhile, AKT1 phosphorylation could also be stimulated by lncRNA MALAT1 based on the recruitment of EZH2 and decrease of BRCA1 expression, thus accelerating inflammatory response to aggravating sepsis (9). Furthermore, in terms of MMP-9, it has been reported that the administration of inhibitors to inhibit the trimeric proteoform of MMP-9 selectively would increase the presence of neutrophils in the inflammation triggering site, suggesting a possible beneficial role of MMP-9 trimers in inhibiting excessive neutrophil migration or anti-inflammatory effect (10). Reckens et al found that MMP-9(-/-) mice developed more severe damage to distant organs during infection, implying that MMP-9 may be significantly involved in the effective host response to E. coli peritonitis (11).…”