Effects of a novel Nodal-targeting monoclonal antibody in melanoma.

// Luigi Strizzi 1,2,* , Annamaria Sandomenico 5,* , Naira V. Margaryan 1 , Annalia Foca 5 , Luca Sanguigno 6 , Thomas M. Bodenstine 1 , Grace S. Chandler 1 , David W. Reed 1 , Alina Gilgur 1 , Elisabeth A. Seftor 1 , Richard E.B. Seftor 1,3 , Zhila Khalkhali-Ellis 1,3,4 , Antonio Leonardi 6 , Menotti Ruvo 5 and Mary J.C. Hendrix 1,4 1 Cancer Biology and Epigenomics Program, Stanley Manne Children’s Research Institute, Ann and Robert H. Lurie Children’s Hospital of Chicago, Chicago, IL, USA 2 Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA 3 Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, IL, USA 4 Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, USA 5 Istituto di Biostrutture e Bioimmagini del CNR and CIRPeB, Universita Federico II di Napoli, Naples, Italy 6 Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Universita Federico II di Napoli, Naples, Italy * These authors have contributed equally to this work Correspondence to: Mary J.C. Hendrix, email: // Menotti Ruvo, email: // Antonio Leonardi, email: // Keywords : Nodal, cancer, antibody, ELISA, therapy Received : July 28, 2015 Accepted : September 24, 2015 Published : October 09, 2015 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.