title: Blockade of MMP14 activity in murine breast carcinomas: implications for macrophages, vessels, and radiotherapy.
abstract: BACKGROUND
  Matrix metalloproteinase (MMP) 14 may mediate tumor progression through vascular and immune-modulatory effects.
  
  
  METHODS
  Orthotopic murine breast tumors (4T1 and E0771 with high and low MMP14 expression, respectively; n = 5-10 per group) were treated with an anti-MMP14 inhibitory antibody (DX-2400), IgG control, fractionated radiation therapy, or their combination. We assessed primary tumor growth, transforming growth factor β (TGFβ) and inducible nitric oxide synthase (iNOS) expression, macrophage phenotype, and vascular parameters. A linear mixed model with repeated observations, with Mann-Whitney or analysis of variance with Bonferroni post hoc adjustment, was used to determine statistical significance. All statistical tests were two-sided.
  
  
  RESULTS
  DX-2400 inhibited tumor growth compared with IgG control treatment, increased macrophage numbers, and shifted the macrophage phenotype towards antitumor M1-like. These effects were associated with a reduction in active TGFβ and SMAD2/3 signaling. DX-2400 also transiently increased iNOS expression and tumor perfusion, reduced tissue hypoxia (median % area: control, 20.2%, interquartile range (IQR) = 6.4%-38.9%; DX-2400: 1.2%, IQR = 0.2%-3.2%, P = .044), and synergistically enhanced radiation therapy (days to grow to 800mm(3): control, 12 days, IQR = 9-13 days; DX-2400 plus radiation, 29 days, IQR = 26-30 days, P < .001) in the 4T1 model. The selective iNOS inhibitor, 1400W, abolished the effects of DX-2400 on vessel perfusion and radiotherapy. On the other hand, DX-2400 was not capable of inducing iNOS expression or synergizing with radiation in E0771 tumors.
  
  
  CONCLUSIONS
  MMP14 blockade decreased immunosuppressive TGFβ, polarized macrophages to an antitumor phenotype, increased iNOS, and improved tumor perfusion, resulting in reduced primary tumor growth and enhanced response to radiation therapy, especially in high MMP14-expressing tumors.
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Eleanor I
authors.initials: EI
authors.lastname: Ager
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Sergey V
authors.initials: SV
authors.lastname: Kozin
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Nathaniel D
authors.initials: ND
authors.lastname: Kirkpatrick
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Giorgio
authors.initials: G
authors.lastname: Seano
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: David P
authors.initials: DP
authors.lastname: Kodack
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Vasileios
authors.initials: V
authors.lastname: Askoxylakis
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Shom
authors.initials: S
authors.lastname: Goel
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Matija
authors.initials: M
authors.lastname: Snuderl
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Alona
authors.initials: A
authors.lastname: Muzikansky
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Dianne M
authors.initials: DM
authors.lastname: Finkelstein
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Dransfield
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Laetitia
authors.initials: L
authors.lastname: Devy
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD).
authors.forename: Yves
authors.initials: Y
authors.lastname: Boucher
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD). jain@steele.mgh.harvard.edu dai@steele.mgh.harvard.edu.
authors.forename: Dai
authors.initials: D
authors.lastname: Fukumura
authors.affiliation: Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston MA (EIA, SVK, NDK, GS, DPK, VA, YH, SG, MS, YB, DF, RKJ); Department of Surgery (Austin Health), University of Melbourne, Studley Road Heidelberg, VIC, Australia (EIA); Novogen, Hornsby, NSW, Australia (EIA); Novartis Institutes for BioMedical Research, Cambridge, MA (NDK); Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Camperdown, NSW, Australia (SG); Department of Biostatistics, Massachusetts General Hospital Biostatistics Center, Boston, Massachusetts (AM, DMF); Department of Pathology, NYU Langone Medical Center and Medical School, New York, NY (MS); Dyax Corp., Burlington, MA (DTD, LD); Departments of Medical Oncology and Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA (SG); Tokai Pharmaceuticals, Inc., Cambridge, MA (DTD); Merck Serono S. A., Geneva, Switzerland (LD). jain@steele.mgh.harvard.edu dai@steele.mgh.harvard.edu.
authors.forename: Rakesh K
authors.initials: RK
authors.lastname: Jain
mesh_terms: D000578:Amidines / Q000494:pharmacology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology* / Q000627:therapeutic use; D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D001596:Benzylamines / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000098:blood supply / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism* / Q000532:radiotherapy*; D045744:Cell Line, Tumor; D019583:Dose Fractionation, Radiation; D004791:Enzyme Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D008264:Macrophages / Q000187:drug effects* / Q000201:enzymology; D008325:Mammary Neoplasms, Experimental; D053511:Matrix Metalloproteinase 14 / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D009389:Neovascularization, Pathologic; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors / Q000187:drug effects* / Q000378:metabolism; D010641:Phenotype; D015398:Signal Transduction / Q000187:drug effects; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D000578:Amidines; D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D001596:Benzylamines; D004791:Enzyme Inhibitors; D007074:Immunoglobulin G; C104820:N-(3-(aminomethyl)benzyl)acetamidine; C497782:SMAD2 protein, human; C497790:SMAD3 protein, human; D051899:Smad2 Protein; D051900:Smad3 Protein; D016212:Transforming Growth Factor beta; C496319:NOS2 protein, human; D052247:Nitric Oxide Synthase Type II; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14