Limits...
ADAM8 expression in invasive breast cancer promotes tumor dissemination and metastasis.

Romagnoli M, Mineva ND, Polmear M, Conrad C, Srinivasan S, Loussouarn D, Barillé-Nion S, Georgakoudi I, Dagg Á, McDermott EW, Duffy MJ, McGowan PM, Schlomann U, Parsons M, Bartsch JW, Sonenshein GE - EMBO Mol Med (2013)

Bottom Line: Circulating tumor cells and brain metastases were also significantly reduced.Furthermore, antibody treatment of established tumors profoundly decreased metastases in a resection model.As a non-essential protein under physiological conditions, ADAM8 represents a promising novel target for treatment of TNBCs, which currently lack targeted therapies and frequently progress with fatal dissemination.

View Article: PubMed Central - PubMed

Affiliation: Department of Developmental, Molecular and Chemical Biology, Tufts University School of Medicine, Boston, MA, USA.

ABSTRACT
The transmembrane metalloprotease-disintegrin ADAM8 mediates cell adhesion and shedding of ligands, receptors and extracellular matrix components. Here, we report that ADAM8 is abundantly expressed in breast tumors and derived metastases compared to normal tissue, especially in triple-negative breast cancers (TNBCs). Furthermore, high ADAM8 levels predicted poor patient outcome. Consistently, ADAM8 promoted an aggressive phenotype of TNBC cells in culture. In a mouse orthotopic model, tumors derived from TNBC cells with ADAM8 knockdown failed to grow beyond a palpable size and displayed poor vascularization. Circulating tumor cells and brain metastases were also significantly reduced. Mechanistically, ADAM8 stimulated both angiogenesis through release of VEGF-A and transendothelial cell migration via β1-integrin activation. In vivo, treatment with an anti-ADAM8 antibody from the time of cell inoculation reduced primary tumor burden and metastases. Furthermore, antibody treatment of established tumors profoundly decreased metastases in a resection model. As a non-essential protein under physiological conditions, ADAM8 represents a promising novel target for treatment of TNBCs, which currently lack targeted therapies and frequently progress with fatal dissemination.

Show MeSH

Related in: MedlinePlus

A–E MDA-MB-231 shCtrl-3 cells were injected into the MFP of female mice. Animals were treated with either 0.5 mg/kg anti-ADAM8 (anti-A8, Mab1031, n = 9) or isotype-matched control (IgG2B, n = 8) in i.p. injection twice weekly. Tumor volume was measured on the indicated days (mean ± s.e.m.). * P = 0.02 (day 11), #P = 0.01 (day 13), †P = 2.1E-4 (day 17), ‡P = 1.5E-4 (day 20), Student's t-test (A). At the end of the experiment, tumors were weighed (mean ± s.e.m.). * P = 5.8E-4, Student's t-test (B) and the presence of brain metastases was examined by fluorescent microscopy (C). Angiogenesis was evaluated by CD31 immunohistochemical staining of tumor sections. Vessel density for each mouse is given as the average number of vessels in 2 slides/tumor (5 peritumoral hot spots/slide) and representative pictures are shown (D). P: Peritumoral area, T: Tumor. Bar: 100 μm. VEGF-A levels in the tumor extracts were determined by WB and normalized to Coomassie staining (E). * P = 0.04 (D), * P = 0.03 (E), Student's t-test.F–G Scheme of experimental design (F). Metastases to the brain and lungs were examined by fluorescent microscopy. Representative images and frequency of metastases (percentage of animals positive per group: IgG2B, n = 8; anti-A8, n = 9) are presented (G). Bar: 250 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3927960&req=5

fig07: A–E MDA-MB-231 shCtrl-3 cells were injected into the MFP of female mice. Animals were treated with either 0.5 mg/kg anti-ADAM8 (anti-A8, Mab1031, n = 9) or isotype-matched control (IgG2B, n = 8) in i.p. injection twice weekly. Tumor volume was measured on the indicated days (mean ± s.e.m.). * P = 0.02 (day 11), #P = 0.01 (day 13), †P = 2.1E-4 (day 17), ‡P = 1.5E-4 (day 20), Student's t-test (A). At the end of the experiment, tumors were weighed (mean ± s.e.m.). * P = 5.8E-4, Student's t-test (B) and the presence of brain metastases was examined by fluorescent microscopy (C). Angiogenesis was evaluated by CD31 immunohistochemical staining of tumor sections. Vessel density for each mouse is given as the average number of vessels in 2 slides/tumor (5 peritumoral hot spots/slide) and representative pictures are shown (D). P: Peritumoral area, T: Tumor. Bar: 100 μm. VEGF-A levels in the tumor extracts were determined by WB and normalized to Coomassie staining (E). * P = 0.04 (D), * P = 0.03 (E), Student's t-test.F–G Scheme of experimental design (F). Metastases to the brain and lungs were examined by fluorescent microscopy. Representative images and frequency of metastases (percentage of animals positive per group: IgG2B, n = 8; anti-A8, n = 9) are presented (G). Bar: 250 μm.

Mentions: To validate ADAM8 as a therapeutic target for TNBC, the Mab1031 ADAM8 antibody was selected for use in an orthotopic model. Mice were treated with either 0.5 mg/kg of Mab1031 antibody ( n = 9) or a control isotype-matched IgG2B ( n = 8) by intra-peritoneal injection twice weekly from the day of implantation of shCtrl-3 cells. After 3 weeks, anti-ADAM8 treatment had significantly decreased primary tumor burden by approximately 50% (Fig 7A) and tumor weight by approximately 36% (Fig 7B). In animals treated with anti-ADAM8, a substantial reduction was also seen in the size and numbers of brain metastases per mouse as well as in the overall frequency (Fig 7C). Surrounding angiogenesis (Fig 7D) and VEGF-A levels in the tumors (Fig 7E) were similarly reduced by the ADAM8 antibody. Using the well-established ADAM8 metalloproteinase substrate CD23 (Koller et al, 2009) in an in vitro analysis, a substantial reduction in ADAM8 activity was observed upon addition of the anti-ADAM8 antibody (supplementary Fig S9).


ADAM8 expression in invasive breast cancer promotes tumor dissemination and metastasis.

Romagnoli M, Mineva ND, Polmear M, Conrad C, Srinivasan S, Loussouarn D, Barillé-Nion S, Georgakoudi I, Dagg Á, McDermott EW, Duffy MJ, McGowan PM, Schlomann U, Parsons M, Bartsch JW, Sonenshein GE - EMBO Mol Med (2013)

A–E MDA-MB-231 shCtrl-3 cells were injected into the MFP of female mice. Animals were treated with either 0.5 mg/kg anti-ADAM8 (anti-A8, Mab1031, n = 9) or isotype-matched control (IgG2B, n = 8) in i.p. injection twice weekly. Tumor volume was measured on the indicated days (mean ± s.e.m.). * P = 0.02 (day 11), #P = 0.01 (day 13), †P = 2.1E-4 (day 17), ‡P = 1.5E-4 (day 20), Student's t-test (A). At the end of the experiment, tumors were weighed (mean ± s.e.m.). * P = 5.8E-4, Student's t-test (B) and the presence of brain metastases was examined by fluorescent microscopy (C). Angiogenesis was evaluated by CD31 immunohistochemical staining of tumor sections. Vessel density for each mouse is given as the average number of vessels in 2 slides/tumor (5 peritumoral hot spots/slide) and representative pictures are shown (D). P: Peritumoral area, T: Tumor. Bar: 100 μm. VEGF-A levels in the tumor extracts were determined by WB and normalized to Coomassie staining (E). * P = 0.04 (D), * P = 0.03 (E), Student's t-test.F–G Scheme of experimental design (F). Metastases to the brain and lungs were examined by fluorescent microscopy. Representative images and frequency of metastases (percentage of animals positive per group: IgG2B, n = 8; anti-A8, n = 9) are presented (G). Bar: 250 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3927960&req=5

fig07: A–E MDA-MB-231 shCtrl-3 cells were injected into the MFP of female mice. Animals were treated with either 0.5 mg/kg anti-ADAM8 (anti-A8, Mab1031, n = 9) or isotype-matched control (IgG2B, n = 8) in i.p. injection twice weekly. Tumor volume was measured on the indicated days (mean ± s.e.m.). * P = 0.02 (day 11), #P = 0.01 (day 13), †P = 2.1E-4 (day 17), ‡P = 1.5E-4 (day 20), Student's t-test (A). At the end of the experiment, tumors were weighed (mean ± s.e.m.). * P = 5.8E-4, Student's t-test (B) and the presence of brain metastases was examined by fluorescent microscopy (C). Angiogenesis was evaluated by CD31 immunohistochemical staining of tumor sections. Vessel density for each mouse is given as the average number of vessels in 2 slides/tumor (5 peritumoral hot spots/slide) and representative pictures are shown (D). P: Peritumoral area, T: Tumor. Bar: 100 μm. VEGF-A levels in the tumor extracts were determined by WB and normalized to Coomassie staining (E). * P = 0.04 (D), * P = 0.03 (E), Student's t-test.F–G Scheme of experimental design (F). Metastases to the brain and lungs were examined by fluorescent microscopy. Representative images and frequency of metastases (percentage of animals positive per group: IgG2B, n = 8; anti-A8, n = 9) are presented (G). Bar: 250 μm.
Mentions: To validate ADAM8 as a therapeutic target for TNBC, the Mab1031 ADAM8 antibody was selected for use in an orthotopic model. Mice were treated with either 0.5 mg/kg of Mab1031 antibody ( n = 9) or a control isotype-matched IgG2B ( n = 8) by intra-peritoneal injection twice weekly from the day of implantation of shCtrl-3 cells. After 3 weeks, anti-ADAM8 treatment had significantly decreased primary tumor burden by approximately 50% (Fig 7A) and tumor weight by approximately 36% (Fig 7B). In animals treated with anti-ADAM8, a substantial reduction was also seen in the size and numbers of brain metastases per mouse as well as in the overall frequency (Fig 7C). Surrounding angiogenesis (Fig 7D) and VEGF-A levels in the tumors (Fig 7E) were similarly reduced by the ADAM8 antibody. Using the well-established ADAM8 metalloproteinase substrate CD23 (Koller et al, 2009) in an in vitro analysis, a substantial reduction in ADAM8 activity was observed upon addition of the anti-ADAM8 antibody (supplementary Fig S9).

Bottom Line: Circulating tumor cells and brain metastases were also significantly reduced.Furthermore, antibody treatment of established tumors profoundly decreased metastases in a resection model.As a non-essential protein under physiological conditions, ADAM8 represents a promising novel target for treatment of TNBCs, which currently lack targeted therapies and frequently progress with fatal dissemination.

View Article: PubMed Central - PubMed

Affiliation: Department of Developmental, Molecular and Chemical Biology, Tufts University School of Medicine, Boston, MA, USA.

ABSTRACT
The transmembrane metalloprotease-disintegrin ADAM8 mediates cell adhesion and shedding of ligands, receptors and extracellular matrix components. Here, we report that ADAM8 is abundantly expressed in breast tumors and derived metastases compared to normal tissue, especially in triple-negative breast cancers (TNBCs). Furthermore, high ADAM8 levels predicted poor patient outcome. Consistently, ADAM8 promoted an aggressive phenotype of TNBC cells in culture. In a mouse orthotopic model, tumors derived from TNBC cells with ADAM8 knockdown failed to grow beyond a palpable size and displayed poor vascularization. Circulating tumor cells and brain metastases were also significantly reduced. Mechanistically, ADAM8 stimulated both angiogenesis through release of VEGF-A and transendothelial cell migration via β1-integrin activation. In vivo, treatment with an anti-ADAM8 antibody from the time of cell inoculation reduced primary tumor burden and metastases. Furthermore, antibody treatment of established tumors profoundly decreased metastases in a resection model. As a non-essential protein under physiological conditions, ADAM8 represents a promising novel target for treatment of TNBCs, which currently lack targeted therapies and frequently progress with fatal dissemination.

Show MeSH
Related in: MedlinePlus