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Journal article
Conformation-Specific Inhibitory Anti-MMP-7 Monoclonal Antibody Sensitizes Pancreatic Ductal Adenocarcinoma Cells to Chemotherapeutic Cell Kill
Cancers, Vol.13(7), 1679
02/Apr/2021
Abstract
Matrix metalloproteases (MMPs) undergo post-translational modifications including pro-domain shedding. The activated forms of these enzymes are effective drug targets, but generating potent biological inhibitors against them remains challenging. We report the generation of anti-MMP-7 inhibitory monoclonal antibody (GSM-192), using an alternating immunization strategy with an active site mimicry antigen and the activated enzyme. Our protocol yielded highly selective anti-MMP-7 monoclonal antibody, which specifically inhibits MMP-7′s enzyme activity with high affinity (IC50 = 132 ± 10 nM). The atomic model of the MMP-7-GSM-192 Fab complex exhibited antibody binding to unique epitopes at the rim of the enzyme active site, sterically preventing entry of substrates into the catalytic cleft. In human PDAC biopsies, tissue staining with GSM-192 showed characteristic spatial distribution of activated MMP-7. Treatment with GSM-192 in vitro induced apoptosis via stabilization of cell surface Fas ligand and retarded cell migration. Co-treatment with GSM-192 and chemotherapeutics, gemcitabine and oxaliplatin elicited a synergistic effect. Our data illustrate the advantage of precisely targeting catalytic MMP-7 mediated disease specific activity.
Details
- Title
- Conformation-Specific Inhibitory Anti-MMP-7 Monoclonal Antibody Sensitizes Pancreatic Ductal Adenocarcinoma Cells to Chemotherapeutic Cell Kill
- Creators
- Vishnu Mohan - 972WIS_INST___120Jean P Gaffney - 972WIS_INST___120Inna Solomonov - 972WIS_INST___120Maxim Levin - 972WIS_INST___120Mordehay Klepfish - 972WIS_INST___120Sophia Akbareian - University of East AngliaBarbara Grünwald - Technical University MunichOrly Dym - 972WIS_INST___589Miriam Eisenstein - 972WIS_INST___111Kenneth H Yu - Memorial Sloan Kettering Cancer CenterDavid P Kelsen - Memorial Sloan Kettering Cancer CenterAchim Krüger - Technical University MunichDylan R Edwards - University of East AngliaIrit Sagi (Corresponding Author) - 972WIS_INST___120
- Resource Type
- Journal article
- Publication Details
- Cancers, Vol.13(7), 1679; 02/Apr/2021
- Number of pages
- 21
- Publisher
- MDPI
- Language
- English
- DOI
- https://doi.org/10.3390/cancers13071679
- Grants
- Grant note
- The authors thank Anjana Shenoy for help in improving the manuscript draft. The authors also thank Assaf Hanuna, Anna Aloshin, Yael Udi, Orith Leitner and Hedva Hamawi for experimental assistance. The image in Figure 5 was created with BioRender.com. Funding - I.S. (Irit Sagi), is an incumbent of the Maurizio Pontecorvo Professorial Chair, and has received funding from the European Union Framework Programme 7 [FP7/2007-2013] “SaveMe, under grant agreement no. 263307, European Union’s Horizon 2020 research and innovation programme (grant agreement No. [801126]), European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No. [695437]), Eu 2020 EDIT consortium, Israeli Science Foundation (1800/19, 1226/13), the USA-Israel Binational Science Foundation (712506-01), The Ambach fund, The Kimmelman center at the WIS, Cynthia and Andrew Adelson fund, Rose Family Fund for Crohn’s and Colitis Research, The Mireille & Murray Steinberg Family Foundation, the German–Israeli Foundation for Scientific Research and Development (GIF), the European Research Council AdG (THZCALORIMETRY-DLV-695437) and The Thompson Family Foundation, Inc. grant “Targeting Pancreas Cancer MSKCC Core Grant”. D.R.E. acknowledges project grant support from Big C (14-09R). Author contributions - I.S. (Irit Sagi), and I.S. (Inna Solomonov), supervised the project, and helped analyze and interpret the data. I.S. (Irit Sagi), V.M. and J.P.G. designed the research. Identification of the antibody was performed by J.P.G. Biochemical characterization was conducted by V.M. and J.P.G., V.M., J.P.G. and M.K. contributed towards the FACS based assays. O.D. conducted crystallization, and solved the crystal structure of GSM-192. M.E. performed protein docking and model analysis. M.L. performed multiple sequence alignment and structural comparison with orthologs/paralogs. V.M. conducted the migration experiments, immunohistochemistry, co-treatment, and synergy determining experiments. S.A., B.G., A.K. and D.R.E. performed lentiviral silencing. K.H.Y. and D.P.K. collected human biopsies. Figures were prepared by V.M. with contributions from I.S. (Irit Sagi), I.S. (Inna Solomonov), S.A., O.D., M.L. and M.E. The manuscript was written by V.M. with contributions from all authors. All authors have read and agreed to the published version of the manuscript.
- Record Identifier
- 993265552403596
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