PROTEOLYTIC ACTIVITY OF MATRIX METALLOPROTEINASE (MMP)-7, MMP-13 AND CATHEPSIN G IMPAIRS INTEGRITY AND FUNCTION OF THERAPEUTIC MONOCLONAL ANTIBODIES USED IN INFLAMMATORY BOWEL DISEASES

CURCIARELLO, RENATA; TREMELLING MARK; DOCENA GUILLERMO; COMPAGNUCCI MALENA; SOBANDE TONI; MACDONALD THOMAS; CURCIARELLO, RENATA; SAMANTHA JONES; ALASTAIR WATSON; BIANCHERI PAOLO

Tucuman

Congreso; LVII REUNIÓN ANUAL SAI 2019; 2019

INTRODUCTION: Biologic therapy is highly effective in inflammatory bowel diseases (IBD). However, a significant proportion of patients fail to respond, and mechanisms underlying primary non-responsiveness are unclear. We have previously observed that proteolytic degradation by Human Neutrophil Elastase (HNE), MMP-3 and MMP-12, which are up-regulated in IBD inflamed mucosa, may contribute to primary non-responsiveness to anti-tumour necrosis factor (TNF)- agents in IBD. We hereby investigated the effect of other proteases on anti-TNF- and anti-integrin biological agents.METHODS: We co-incubated increasing concentrations of activated recombinant human MMP-7, MMP-13, and Cathepsin G with anti-TNF- agents Infliximab (IFX), Adalimumab (ADA), and Etanercept (ETA), or with anti-integrin agent Vedolizumab (Vedo). Digestions with pepsin and papain were performed as control for immunoglobulin´s proteolytic cleavage. We analysed the cleavage reaction products by western blot. The TNF-α neutralising ability of degraded biological agents was evaluated using a reporter cell line.RESULTS: IFX, ADA and Vedo were not degraded by MMP-7, MMP-13, nor Cathepsin G. ETA was degraded in a concentration-dependent manner by MMP-7 and MMP-13, but not by Cathepsin G. Degradation by MMP-7 and MMP-13 did not significantly impair the ability of Etanercept to neutralise soluble TNF-. Even though pepsin cleavage site in the lower hinge region of IgG is similar for MMP-7, pepsin digestion impaired TNF neutralisation ability of IFX and ETA.CONCLUSIONS: Despite being known to cleave immunoglobulins, MMP-7 and MMP-13 did not impair the integrity and function of Infliximab and Adalimumab. In keeping with our previous findings, Etanercept appears to be highly susceptible to proteases.