A bi-specific inhibitor targeting IL-17A and MMP-9 reduces invasion and motility in MDA-MB-231 cells

Abstract

The cytokine IL-17A is associated with the progression of various cancers, but little is known about the molecular cross-talk between IL-17A and other tumor-promoting factors. Previous studies have shown that the IL-17A-mediated invasion of breast cancer cells can be inhibited by selective antagonists of the matrix metalloproteinase 9 (MMP-9), suggesting that the cross-talk between IL-17A and MMP-9 may promote cancer invasiveness and metastasis. Here, we present a novel strategy for developing cancer therapeutics, based on the simultaneous binding and inhibition of both IL-17A and MMP-9. To this end, we use a bi-specific heterodimeric fusion protein, comprising a natural inhibitor of MMPs (N-TIMP2) fused with an engineered extracellular domain (V3) of the IL-17A receptor. We show that, as compared with the mono-specific inhibitors of IL-17A (V3) and MMP-9 (N-TIMP2), the engineered bi-specific fusion protein inhibits both MMP-9 activation and IL-17A-induced cytokine secretion from fibroblasts and exhibits a synergistic inhibition of both the migration and invasion of breast cancer cells. Our findings demonstrate, for the first time, that dual targeting of inflammatory (IL-17A) and extracellular matrix remodeling (MMP) pathways can potentially be used as a novel therapeutic approach against cancer. Moreover, the platform developed here for generating the bi-specific IL-17A/MMP-9 inhibitor can be utilized for generating bi-specific inhibitors for other cytokines and MMPs.

Metalloproteinases and Their Inhibitors: Potential for the Development of New Therapeutics

“…The isolated TIMP-1 mutants showed an order of magnitude improvement in binding to MMP-3 catalytic domain (MMP-3cd). Although most of the focus for engineering TIMPs were focused on the N-terminal/inhibitory domain of TIMPs [103,251,252], it was shown that cooperation between Nand C-terminal domains of TIMP-1 improved binding to the target MMP-3.…”

“…The TIMP interaction surface with MMPs contains six main regions (CTC motif at the N-terminus, AB-loop, Cconnector loop, GH-loop, EF-loop, and MTL-loop), which resemble monoclonal antibody (mAb) IgG The isolated TIMP-1 mutants showed an order of magnitude improvement in binding to MMP-3 catalytic domain (MMP-3cd). Although most of the focus for engineering TIMPs were focused on the N-terminal/inhibitory domain of TIMPs [103,251,252], it was shown that cooperation between N-and C-terminal domains of TIMP-1 improved binding to the target MMP-3.…”

The role of Th-17 cells and IL-17 in the metastatic spread of breast cancer: As a means of prognosis and therapeutic target

“…Thus, cancer develops after migration to other anatomic sites, which are called secondary tumors ( 96 ). NF-kB key transcription factor plays a role in the expression and activity of MMPs ( 16 , 17 , 97 ). This, in turn, defines as many of the effects of IL-17A that are correlated with the TRAF-6-mediated activation of NF-kB.…”

“…As discussed earlier, IL-17A mediates cancer cell invasiveness and metastasis via MMP-2, MMP-9, and MMP-13. Furthermore, IL-17A stimulates MMP-9 mRNA expression, and MMP-9 inhibitors can inhibit the IL-17A-dependent invasion and metastasis of BCCs ( 17 ). The relation between IL-17A and its downstream MMP activity and breast cancer metastasis through MAPK and NF-Kb suggests the possibility of various strategies connected with blocking these checkpoints and kinase enzyme activity.…”

“…For the effective migration and metastasis of breast cancer cells in the vascular or lymphatic drainage system, chemical mediators such as calcium-dependent zinc-containing endopeptidases like MMPs must be required for the degradation of the ECM as well as VEGF and IL-8 for vascularization during intravasation and extravasation processes, respectively, and reach to the bone ( 15 ). A study showed that the expression and activation of MMPs are mediated through TNF-α and IL-1 secreted by tumor cells, and IL-17A secreted from the microenvironment plays a role on the regulation of different MMPs ( 16 , 17 ). There are five major classes of MMPs depending on their function and the substrates that they digest, including matrilysins (MMP-7 and MMP-26), collagenases (MMP-1, MMP-8, and MMP-13), stromelysins (MMP-3, MMP-10, and MMP-11), gelatinase (MMP-2 and MMP-9), and membrane-associated metalloproteinases (MMP-14, MMP-15, MMP-16, MMP-17, MMP-23A/B, MMP-24, and MMP-25) ( 18 , 19 ).…”