title: Effects of a novel Nodal-targeting monoclonal antibody in melanoma.
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.
authors.affiliation: Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.|Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
authors.forename: Luigi
authors.initials: L
authors.lastname: Strizzi
authors.affiliation: Istituto di Biostrutture e Bioimmagini del CNR and CIRPeB, Università Federico II di Napoli, Naples, Italy.
authors.forename: Annamaria
authors.initials: A
authors.lastname: Sandomenico
authors.affiliation: Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.
authors.forename: Naira V
authors.initials: NV
authors.lastname: Margaryan
authors.affiliation: Istituto di Biostrutture e Bioimmagini del CNR and CIRPeB, Università Federico II di Napoli, Naples, Italy.
authors.forename: Annalia
authors.initials: A
authors.lastname: Focà
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università Federico II di Napoli, Naples, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Sanguigno
authors.affiliation: Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Bodenstine
authors.affiliation: Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.
authors.forename: Grace S
authors.initials: GS
authors.lastname: Chandler
authors.affiliation: Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.
authors.forename: David W
authors.initials: DW
authors.lastname: Reed
authors.affiliation: Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.
authors.forename: Alina
authors.initials: A
authors.lastname: Gilgur
authors.affiliation: Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.
authors.forename: Elisabeth A
authors.initials: EA
authors.lastname: Seftor
authors.affiliation: Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.|Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
authors.forename: Richard E B
authors.initials: RE
authors.lastname: Seftor
authors.affiliation: Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.|Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.|Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
authors.forename: Zhila
authors.initials: Z
authors.lastname: Khalkhali-Ellis
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università Federico II di Napoli, Naples, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Leonardi
authors.affiliation: Istituto di Biostrutture e Bioimmagini del CNR and CIRPeB, Università Federico II di Napoli, Naples, Italy.
authors.forename: Menotti
authors.initials: M
authors.lastname: Ruvo
authors.affiliation: Cancer Biology and Epigenomics Program, Stanley Manne Children's Research Institute, Ann and Robert H. Lurie Children's Hospital of Chicago, Chicago, IL, USA.|Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
authors.forename: Mary J C
authors.initials: MJ
authors.lastname: Hendrix
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001943:Breast Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D056744:Cyclin B1 / Q000096:biosynthesis; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay; D048049:Extracellular Signal-Regulated MAP Kinases / Q000096:biosynthesis; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D008545:Melanoma / Q000473:pathology*; D051379:Mice; D055457:Nodal Protein / Q000037:antagonists & inhibitors* / Q000097:blood / Q000276:immunology; D010091:Oximes / Q000494:pharmacology; D048493:Proto-Oncogene Proteins B-raf / Q000037:antagonists & inhibitors* / Q000235:genetics; D051899:Smad2 Protein / Q000096:biosynthesis; D020349:Surface Plasmon Resonance
chemical_list: D000911:Antibodies, Monoclonal; C534699:CCNB1 protein, human; D056744:Cyclin B1; D007093:Imidazoles; C526007:NODAL protein, human; D055457:Nodal Protein; D010091:Oximes; C497782:SMAD2 protein, human; D051899:Smad2 Protein; D050760:Cyclin-Dependent Kinase Inhibitor p27; D048493:Proto-Oncogene Proteins B-raf; D048049:Extracellular Signal-Regulated MAP Kinases; C561627:dabrafenib
keywords: ELISA; Nodal; antibody; cancer; therapy