A novel immunotherapy targeting MMP-14 limits hypoxia, immune suppression and metastasis in triple-negative breast cancer models

Abstract

Matrix metalloproteinase-14 (MMP-14) is a clinically relevant target in metastatic cancers due to its role in tumor progression and metastasis. Since active MMP-14 is localized on the cell surface, it is amenable to antibody-mediated blockade in cancer, and here we describe our efforts to develop novel inhibitory anti-MMP-14 antibodies. A phage-displayed synthetic humanized Fab library was screened against the extracellular domain of MMP-14 and a panel of MMP14-specific Fabs were identified. A lead antibody that inhibits the catalytic domain of MMP-14 (Fab 3369) was identified and treatment of MDA-MB-231 breast cancer cells with Fab 3369 led to significant loss of extracellular matrix degradation and cell invasion abilities. In mammary orthotopic tumor xenograft assays, MMP-14 blockade by IgG 3369 limited tumor growth and metastasis. Analysis of tumor tissue sections revealed that MMP-14 blockade limited tumor neoangiogenesis and hypoxia. Similar effects of MMP-14 blockade in syngeneic 4T1 mammary tumors were observed, along with increased detection of cytotoxic immune cell markers. In conclusion, we show that immunotherapies targeting MMP-14 can limit immune suppression, tumor progression, and metastasis in triple-negative breast cancer.

Epitope‐specific affinity maturation improved stability of potent protease inhibitory antibodies

“…However, the high similarity of protein folding and catalytic chemistry among MMP family members presents a daunting challenge for the generation of highly selective compound inhibitors (Turk, ). Our studies (Lopez, Nam, Kaihara, Mustafa & Ge, ; Nam, Fang, Rodriguez, Lopez, & Ge, ; Nam, Rodriguez, Remacle, Strongin, & Ge, ), among others (Appleby et al, ; Devy et al, ; Ling et al, ; Udi et al, ), demonstrated the feasibility that antibody‐based inhibitors could exhibit the desired high selectivity. Particularly, Fab3A2 with 4.8 nM affinity, 9.7 nM potency, and high selectivity toward MMP‐14 was isolated from a library containing ultralong CDR H3s (Nam et al, ).…”

“…Our studies (Lopez, Nam, Kaihara, Mustafa & Ge, 2017;Nam, Fang, Rodriguez, Lopez, & Ge, 2017;Nam, Rodriguez, Remacle, Strongin, & Ge, 2016), among others (Appleby et al, 2017;Devy et al, 2009;Ling et al, 2017;Udi et al, 2015), demonstrated the feasibility that antibody-based inhibitors could exhibit the desired high selectivity.…”

Characterization and regulation of MT1‐MMP cell surface‐associated activity

“…However, gelatin is a substrate for multiple proteases, and MMPs were expressed on the surface of yeast cells (Diehl et al, 2011), which are quite different from mammalian cell surfaces. As several selective and/or secondary binding site (exosite) MT1-MMP inhibitors have been described (Levin, Udi, Solomonov, & Sagi, 2017;Ling et al, 2017;Pahwa et al, 2014;Santamaria & de Groot, 2018), the evaluation of activity at the cell surface will allow for examination of such inhibitors in a more native-like environment. This will also facilitate the development of inhibitors that may act indirectly on MT1-MMP activity.…”

Single peptides and combination modalities for triple negative breast cancer

“…Other inhibitory molecules include Fab R2C7 which is highly selective for MMP‐14 (Lopez, Nam, Kaihara, Mustafa, & Ge, ). Another MMP‐14 inhibitor that inhibits the catalytic domain of MMP‐14, Fab 3369, prevents the ECM degradation by MMP‐14 invasion of MDA‐MB‐231 cells (Ling et al, ). The blockade of MMP‐14 has been shown to overcome the immunosuppressive TME in metastatic cases BC (Li et al, ).…”

“…One of the MMP family members, MMP-14 can degrade collagen, which is required for invadopodia formation (Leong et al, 2014). Targeting the metastasis early events for example the degradation step of ECM and the cancer cells invasion, may proved beneficial effects in TNBC (Ling et al, 2017).…”