TY - JOUR
T1 - Targeting the MMP-14/MMP-2/integrin v3 axis with multispecific N-TIMP2– based antagonists for cancer therapy
AU - Yosef, Gal
AU - Arkadash, Valeria
AU - Papo, Niv
N1 - Publisher Copyright: © 2018 Yosef et al. Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2018/8/24
Y1 - 2018/8/24
N2 - The pathophysiological functions of the signaling molecules matrix metalloproteinase-14 (MMP-14) and integrinv3 in various types of cancer are believed to derive from their collaborative activity in promoting invasion, metastasis, and angiogenesis, as shown in vitro and in vivo. The two effectors act in concert in a cell-specific manner through the localization of pro-MMP-2 to the cell surface, where it is processed to intermediate and matured MMP-2. The matured MMP-2 product is localized to the cell surface via its binding to integrinv3. The MMP-14/MMP-2/integrinv3 axis thus constitutes an attractive putative target for therapeutic interventions, but the development of inhibitors that target this axis remains an unfulfilled task. To address the lack of such multitarget inhibitors, we have established a combinatorial approach that is based on flow cytometry screening of a yeast-displayed N-TIMP2 (N-terminal domain variant of tissue inhibitor of metalloproteinase-2) mutant library. On the basis of this screening, we generated protein monomers and a heterodimer that contain monovalent and bivalent binding epitopes to MMP-14 and integrinv3. Among these proteins, the bi-specific heterodimer, which bound strongly to both MMP-14 and integrinv3, exhibited superior ability to inhibit MMP-2 activation and displayed the highest inhibitory activity in cell-based models of a MMP-14-, MMP-2-, and integrinv3-dependent glioblastoma and of endothelial cell invasiveness and endothelial capillary tube formation. These assays enabled us to show the superiority of the combined target effects of the inhibitors and to investigate separately the role each of the three signaling molecules in various malignant processes.
AB - The pathophysiological functions of the signaling molecules matrix metalloproteinase-14 (MMP-14) and integrinv3 in various types of cancer are believed to derive from their collaborative activity in promoting invasion, metastasis, and angiogenesis, as shown in vitro and in vivo. The two effectors act in concert in a cell-specific manner through the localization of pro-MMP-2 to the cell surface, where it is processed to intermediate and matured MMP-2. The matured MMP-2 product is localized to the cell surface via its binding to integrinv3. The MMP-14/MMP-2/integrinv3 axis thus constitutes an attractive putative target for therapeutic interventions, but the development of inhibitors that target this axis remains an unfulfilled task. To address the lack of such multitarget inhibitors, we have established a combinatorial approach that is based on flow cytometry screening of a yeast-displayed N-TIMP2 (N-terminal domain variant of tissue inhibitor of metalloproteinase-2) mutant library. On the basis of this screening, we generated protein monomers and a heterodimer that contain monovalent and bivalent binding epitopes to MMP-14 and integrinv3. Among these proteins, the bi-specific heterodimer, which bound strongly to both MMP-14 and integrinv3, exhibited superior ability to inhibit MMP-2 activation and displayed the highest inhibitory activity in cell-based models of a MMP-14-, MMP-2-, and integrinv3-dependent glioblastoma and of endothelial cell invasiveness and endothelial capillary tube formation. These assays enabled us to show the superiority of the combined target effects of the inhibitors and to investigate separately the role each of the three signaling molecules in various malignant processes.
UR - http://www.scopus.com/inward/record.url?scp=85052144946&partnerID=8YFLogxK
U2 - https://doi.org/10.1074/jbc.RA118.004406
DO - https://doi.org/10.1074/jbc.RA118.004406
M3 - Article
SN - 0021-9258
VL - 293
SP - 13310
EP - 13326
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 34
ER -