Effects of ADAM10 and ADAM17 Inhibitors on Natural Killer Cell Expansion and Antibody-dependent Cellular Cytotoxicity Against Breast Cancer Cells In Vitro

Anticancer Res. 2017 Oct;37(10):5507-5513. doi: 10.21873/anticanres.11981.

Abstract

Background/aim: The inhibition of a disintegrin and metalloproteinase (ADAM) has the potential to become a novel approach for natural killer (NK) cell-based cancer immunotherapy. Thus, the aim of this study was to investigate the influence of ADAM10 and ADAM17 inhibitors on expanded NK cell to enhance antibody-dependent cellular cytotoxicity (ADCC) in breast cancer cell lines.

Materials and methods: NK cells were expanded in medium supplemented with an ADAM10 or ADAM17 inhibitor to prevent the shedding of soluble CD16/FcγRIII. The expression level of CD16 and production of interferon-gamma (IFN-γ) was detected by flow cytometry using specific antibodies. ADCC activity of expanded NK cells was estimated in trastuzumab treated breast cancer cell lines such as MCF-7, MDA-MB-231, SKBR3, and BT-474 cells.

Results: The ADAM17 inhibitor increased the purity of expanded NK cells to 90% after 14 days at 5 and 10 μM in vitro (p=0.043). However, the expansion rate of NK cells was decreased at 10 μM of the ADAM 17 inhibitor (p=0.043). Inhibition of ADAM10 suppressed the expansion of NK cells, although the NK purity was increased at 1 μM of the inhibitor. The expression of CD16 was significantly increased at 1 and 5 μM of the ADAM17 inhibitor (p=0.046, 0.028, respectively) during the culturing period. Inhibition of ADAM10 reduced the expression of CD16 on NK cells. The cytotoxic activity of the ADAM17 inhibitor treated NK cells against MCF-7 (p=0.039) and BT-474 (p=0.027) cells was significantly elevated. The ADCC activity of NK cells treated with 5 μM of ADAM17 inhibitor was significantly increased against SKBR-3 and BT-474 (p=0.027). Inhibition of ADAM17 increased the production of IFN-γ in expanded NK cells.

Conclusion: The inhibition of ADAM17 enhanced the purity of expanded NK cells and the ADCC activity of these cells against trastuzumab treated breast cancer cell lines.

Keywords: ADAM inhibitor; CD16 receptor; NK cell; breast neoplasms.

MeSH terms

  • ADAM10 Protein / antagonists & inhibitors*
  • ADAM10 Protein / metabolism
  • ADAM17 Protein / antagonists & inhibitors*
  • ADAM17 Protein / metabolism
  • Amyloid Precursor Protein Secretases / antagonists & inhibitors*
  • Amyloid Precursor Protein Secretases / metabolism
  • Antibody-Dependent Cell Cytotoxicity / drug effects*
  • Antineoplastic Agents / pharmacology*
  • Antineoplastic Combined Chemotherapy Protocols / pharmacology
  • Breast Neoplasms / drug therapy*
  • Breast Neoplasms / enzymology
  • Breast Neoplasms / immunology
  • Breast Neoplasms / pathology
  • Cell Proliferation / drug effects*
  • Coculture Techniques
  • Dose-Response Relationship, Drug
  • Female
  • GPI-Linked Proteins / metabolism
  • Humans
  • Interferon-gamma / metabolism
  • Killer Cells, Natural / drug effects*
  • Killer Cells, Natural / enzymology
  • Killer Cells, Natural / immunology
  • Lymphocyte Activation / drug effects*
  • MCF-7 Cells
  • Membrane Proteins / antagonists & inhibitors*
  • Membrane Proteins / metabolism
  • Protease Inhibitors / pharmacology*
  • Receptors, IgG / metabolism
  • Time Factors
  • Trastuzumab / pharmacology
  • Tumor Microenvironment

Substances

  • Antineoplastic Agents
  • FCGR3B protein, human
  • GPI-Linked Proteins
  • IFNG protein, human
  • Membrane Proteins
  • Protease Inhibitors
  • Receptors, IgG
  • Interferon-gamma
  • Amyloid Precursor Protein Secretases
  • ADAM10 Protein
  • ADAM10 protein, human
  • ADAM17 Protein
  • ADAM17 protein, human
  • Trastuzumab