Selective Inhibition of Matrix Metalloproteinase-14 Blocks Tumor Growth, Invasion, and Angiogenesis

Devy, Laetitia; Huang, Lili; Naa, Laurent; Yanamandra, Niranjan; Pieters, Henk; Frans, Nicolas; Chang, Edward; Qian, Tao; Vanhove, Marc; Lejeune, Aude; Gool, Reinoud van; Sexton, Daniel J.; Kuang, Guannan; Rank, Douglas; Hogan, Sean A.; Pazmany, Csaba; Lu, Yu; Schoonbroodt, Sonia; Nixon, Andrew E.; Ladner, Robert C.; Hoet, Rene; Henderikx, Paula; TenHoor, Christopher; Rabbani, Shafaat A.; Valentino, Maria Luisa; Wood, Clive R.; Dransfield, Daniel T.

doi:10.1158/0008-5472.can-08-3255

Cited by 312 publications

(299 citation statements)

References 34 publications

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“…It was reported that a selective MT1-MMP inhibitor almost completely blocked proMMP-2 processing in HUVECs (28). However, MT1-MMP depletion was not relevant to MMP-2 depletion in RECK-depleted HUVECs in our experiment.…”

Section: Discussioncontrasting

confidence: 61%

The β1-Integrin–Dependent Function of RECK in Physiologic and Tumor Angiogenesis

Miki

Shamma

Kitajima

et al. 2010

Molecular Cancer Research

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Vascular endothelial cells produce considerable amounts of matrix metalloproteinases (MMP), including MMP-2, MMP-9, and membrane type 1 (MT1)-MMP. However, little is known about the regulatory mechanisms of these protease activities exhibited during vascular development. A glycosylphosphatidylinositol-anchored glycoprotein, reversion-inducing cysteine-rich protein with Kazal motifs (RECK), has been shown to attenuate MMP-2 maturation by directly interacting with MT1-MMP. Here, we show that an angiogenic factor angiopoietin-1 induces RECK expression in human umbilical vein endothelial cells (HUVEC), and RECK depletion in these cells results in defective vascular tube formation and cellular senescence. We further observed that RECK depletion downregulates β1-integrin activation, which was associated with decreased autophosphorylation of focal adhesion kinase and increased expression of a cyclin-dependent kinase inhibitor p21 CIP1 . In agreement, significant downregulation of β1-integrin activity was observed in vascular endothelial cells in Reck−/− mouse embryos. In HUVECs, specific inhibition of MMP-2 significantly antagonized the effect of RECK depletion on β1-integrin signaling, cell proliferation, and tube elongation. Furthermore, we observed that hypervascular tumor-derived cell lines can induce high RECK expression in convoluted vascular endothelial cells, and this in turn supports tumor growth. Targeting RECK specifically in tumor-associated vascular endothelial cells resulted in tumor regression. Therefore, we propose that RECK in tumor vascular endothelial cells can be an interesting target of cancer treatment via abortion of tumor angiogenesis.

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Section: Discussioncontrasting

confidence: 61%

The β1-Integrin–Dependent Function of RECK in Physiologic and Tumor Angiogenesis

Miki

Shamma

Kitajima

et al. 2010

Molecular Cancer Research

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“…The presence of 70% of the inhibitory clones among the binders is remarkably high, especially if compared with previous studies of the inhibitory antibodies (hit rates of 2.5% and 17%) (17,36). The inhibition IC 50 values of Fabs 3A2, 3E2, and 3D9 were 9.7 nM, 42 nM, and 61 nM, respectively, and the IC 50 values of three additional Fabs were in 240-420 nM range (Fig.…”

Section: Significancementioning

confidence: 59%

“…The inhibitor/binder ratio was significantly improved when a specific selection procedure was added to phage panning (17). Thus, the use of TIMP-2, a natural protein inhibitor of MMP-14, as a competitive eluent of the antigen-binding clones from the MMP-14 bait led to the discovery of 12 inhibitory constructs from 70 affinity clones (17% hit rate).…”

Section: Discussionmentioning

confidence: 99%

“…As a result, broad-spectrum hydroxamates failed in cancer clinical trials due to their low overall efficacy and side effects (13). Alternatively, antibody-based MMP inhibitors are emerging as both research tools and potential therapeutic agents (10,(17)(18)(19)(20)(21) because of (i) high affinity and specificity due to the large antigen-antibody interaction area and multiple complementarity-determining regions (CDRs), (ii) long half-life and well-defined action mechanisms, (iii) low immunogenicity and toxicity, and (iv) multiple MMPs potentially targetable by antibodies (9).…”

mentioning

confidence: 99%

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Active-site MMP-selective antibody inhibitors discovered from convex paratope synthetic libraries

Nam

Rodríguez

Remacle

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

110

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Proteases are frequent pharmacological targets, and their inhibitors are valuable drugs in multiple pathologies. The catalytic mechanism and the active-site fold, however, are largely conserved among the protease classes, making the development of the selective inhibitors exceedingly challenging. In our departure from the conventional strategies, we reviewed the structure of known camelid inhibitory antibodies, which block enzyme activities via their unusually long, convex-shaped paratopes. We synthesized the human Fab antibody library (over 1.25 × 10 9 individual variants) that carried the extended, 23-to 27-residue, complementarity-determining region (CDR)-H3 segments. As a proof of principle, we used the catalytic domain of matrix metalloproteinase-14 (MMP-14), a promalignant protease and a drug target in cancer, as bait. In our screens, we identified 20 binders, of which 14 performed as potent and selective inhibitors of MMP-14 rather than as broad-specificity antagonists. Specifically, Fab 3A2 bound to MMP-14 in the vicinity of the active pocket with a high 4.8 nM affinity and was similarly efficient (9.7 nM) in inhibiting the protease cleavage activity. We suggest that the convex paratope antibody libraries described here could be readily generalized to facilitate the design of the antibody inhibitors to many additional enzymes. family members control various physiological and pathological processes. Multiple diseases are associated with altered MMP expression and aberrant proteolysis, including cancer (1), wound healing (2), inflammatory diseases (3, 4), neurological pain (5, 6), and hypertension (7). There is consensus among researchers that the individual MMPs are promising drug targets in diversified pathologies and that inhibitor specificity is required for selective and successful MMP therapies (8-10).However, achieving target specificity and selectivity in small-molecule MMP inhibitors is remarkably challenging (11,12). Because the catalytic mechanism and catalytic domain fold are conserved among the MMP/ADAM (a disintegrin and metalloproteinase)/ ADAMTS (ADAM with thrombospondin motifs) superfamily members, the available small-molecule inhibitors (most frequently, active-site zinc-chelating hydroxamates) target multiple proteinases, resulting in off-target side effects (8,(12)(13)(14). This aspect is problematic, given that some MMPs (e.g., MMP-14) are always protumorigenic, whereas some other MMPs are antitumorigenic in certain cancer microenvironments (15, 16). As a result, broad-spectrum hydroxamates failed in cancer clinical trials due to their low overall efficacy and side effects (13). Alternatively, antibody-based MMP inhibitors are emerging as both research tools and potential therapeutic agents (10, 17-21) because of (i) high affinity and specificity due to the large antigen-antibody interaction area and multiple complementarity-determining regions (CDRs), (ii) long half-life and well-defined action mechanisms, (iii) low immunogenicity and toxicity, and (iv) multiple MMPs potentially targe...

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“…30,31 To determine Figure 5c). The data presented above show that the second-generation MLVshRNA vectors are able to distribute shRNA to cultivated cells.…”

Section: Resultsmentioning

confidence: 99%

RNAi-mediated gene silencing in tumour tissue using replication-competent retroviral vectors

et al. 2011

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RNAi represents a powerful technology to specifically downregulate the expression of target genes. For cancer research and therapy, an efficient in vivo delivery system is supposed to distribute RNAi to all tumour cells upon systemic administration. We present replication-competent murine leukaemia virus (MLV) vectors, which deliver RNAi to tumour tissue upon tail vein injection. In HT1080 cells stably expressing GFP or luciferase, GFP expression was suppressed by more than 80% and luciferase (luc) activity by more than 90%, even when only 0.1% of the cells were initially infected with reporter gene specific vectors. To demonstrate its potential, PLK1-and MMP14-specific small hairpin RNA expression cassettes were applied in the system. Upon infection, PLK1 and MMP14 levels were reduced on mRNA and protein level. MLV-shPLK1-infected cells were arrested in the G2-phase and underwent apoptosis. MLV-shMMP14-infected cells showed reduced MMP2 activity, as well as substantially reduced invasion and tumour growth. In vivo, MLV-shLuc silenced luc expression in HT1080-luc tumour tissue by more than 80% and MLV-shPLK1 reduced tumour growth substantially, demonstrating the therapeutic relevance of this system. This RNAi vector system allows long-term downregulation of target gene expression as well as efficient delivery to and distribution throughout tumour tissue in vivo.

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Selective Inhibition of Matrix Metalloproteinase-14 Blocks Tumor Growth, Invasion, and Angiogenesis

Abstract: Inhibition of specific matrix metalloproteinases (MMP) is an attractive noncytotoxic approach to cancer therapy.

Cited by 312 publications

References 34 publications

The β1-Integrin–Dependent Function of RECK in Physiologic and Tumor Angiogenesis

The β1-Integrin–Dependent Function of RECK in Physiologic and Tumor Angiogenesis

Active-site MMP-selective antibody inhibitors discovered from convex paratope synthetic libraries

RNAi-mediated gene silencing in tumour tissue using replication-competent retroviral vectors

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