Effects of a novel Nodal-targeting monoclonal antibody in melanoma

Oncotarget. 2015 Oct 27;6(33):34071-86. doi: 10.18632/oncotarget.6049.

Abstract

Nodal is highly expressed in various human malignancies, thus supporting the rationale for exploring Nodal as a therapeutic target. Here, we describe the effects of a novel monoclonal antibody (mAb), 3D1, raised against human Nodal. In vitro treatment of C8161 human melanoma cells with 3D1 mAb shows reductions in anchorage-independent growth and vasculogenic network formation. 3D1 treated cells also show decreases of Nodal and downstream signaling molecules, P-Smad2 and P-ERK and of P-H3 and CyclinB1, with an increase in p27. Similar effects were previously reported in human breast cancer cells where Nodal expression was generally down-regulated; following 3D1 mAb treatment, both Nodal and P-H3 levels are reduced. Noteworthy is the reduced growth of human melanoma xenografts in Nude mice treated with 3D1 mAb, where immunostaining of representative tumor sections show diminished P-Smad2 expression. Similar effects both in vitro and in vivo were observed in 3D1 treated A375SM melanoma cells harboring the active BRAF(V600E) mutation compared to treatments with IgG control or a BRAF inhibitor, dabrafenib. Finally, we describe a 3D1-based ELISA for the detection of Nodal in serum samples from cancer patients. These data suggest the potential of 3D1 mAb for selecting and targeting Nodal expressing cancers.

Keywords: ELISA; Nodal; antibody; cancer; therapy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Breast Neoplasms / pathology*
  • Cell Line, Tumor
  • Cyclin B1 / biosynthesis
  • Cyclin-Dependent Kinase Inhibitor p27 / biosynthesis
  • Enzyme-Linked Immunosorbent Assay
  • Extracellular Signal-Regulated MAP Kinases / biosynthesis
  • Female
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Imidazoles / pharmacology
  • Melanoma / pathology*
  • Mice
  • Nodal Protein / antagonists & inhibitors*
  • Nodal Protein / blood
  • Nodal Protein / immunology
  • Oximes / pharmacology
  • Proto-Oncogene Proteins B-raf / antagonists & inhibitors*
  • Proto-Oncogene Proteins B-raf / genetics
  • Smad2 Protein / biosynthesis
  • Surface Plasmon Resonance

Substances

  • Antibodies, Monoclonal
  • CCNB1 protein, human
  • Cyclin B1
  • Imidazoles
  • NODAL protein, human
  • Nodal Protein
  • Oximes
  • SMAD2 protein, human
  • Smad2 Protein
  • Cyclin-Dependent Kinase Inhibitor p27
  • Proto-Oncogene Proteins B-raf
  • Extracellular Signal-Regulated MAP Kinases
  • dabrafenib