title: Synergistic anti-tumor activity and inhibition of angiogenesis by cotargeting of oncogenic and death receptor pathways in human melanoma.
abstract: Improving treatment of advanced melanoma may require the development of effective strategies to overcome resistance to different anti-tumor agents and to counteract relevant pro-tumoral mechanisms in the microenvironment. Here we provide preclinical evidence that these goals can be achieved in most melanomas, by co-targeting of oncogenic and death receptor pathways, and independently of their BRAF, NRAS, p53 and PTEN status. In 49 melanoma cell lines, we found independent susceptibility profiles for response to the MEK1/2 inhibitor AZD6244, the PI3K/mTOR inhibitor BEZ235 and the death receptor ligand TRAIL, supporting the rationale for their association. Drug interaction analysis indicated that a strong synergistic anti-tumor activity could be achieved by the three agents and the AZD6244-TRAIL association on 20/21 melanomas, including cell lines resistant to the inhibitors or to TRAIL. Mechanistically, synergy was explained by enhanced induction of caspase-dependent apoptosis, mitochondrial depolarization and modulation of key regulators of extrinsic and intrinsic cell death pathways, including c-FLIP, BIM, BAX, clusterin, Mcl-1 and several IAP family members. Moreover, silencing experiments confirmed the central role of Apollon downmodulation in promoting the apoptotic response of melanoma cells to the combinatorial treatments. In SCID mice, the AZD6244-TRAIL association induced significant growth inhibition of a tumor resistant to TRAIL and poorly responsive to AZD6244, with no detectable adverse events on body weight and tissue histology. Reduction in tumor volume was associated not only with promotion of tumor apoptosis but also with suppression of the pro-angiogenic molecules HIF1α, VEGFα, IL-8 and TGFβ1 and with inhibition of tumor angiogenesis. These results suggest that synergistic co-targeting of oncogenic and death receptor pathways can not only overcome melanoma resistance to different anti-tumor agents in vitro but can also promote pro-apoptotic effects and inhibition of tumor angiogenesis in vivo.
authors.affiliation: Human Tumors Immunobiology Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Grazia
authors.affiliation: Human Tumors Immunobiology Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Vegetti
authors.affiliation: San Raffaele Scientific Institute, URI, Milan, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Benigni
authors.affiliation: Human Tumors Immunobiology Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Penna
authors.affiliation: Human Tumors Immunobiology Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: V
authors.initials: V
authors.lastname: Perotti
authors.affiliation: Human Tumors Immunobiology Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Tassi
authors.affiliation: Human Tumors Immunobiology Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Bersani
authors.affiliation: Human Tumors Immunobiology Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Nicolini
authors.affiliation: Functional Genomics Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Canevari
authors.affiliation: 1] Department of Oncology and Hematology, Humanitas Cancer Center, Humanitas Clinical and Research Center, Rozzano, Italy [2] Department of Medical Biotechnology and Translational Medicine, University of Milan, Milan, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Carlo-Stella
authors.affiliation: Medical Oncology Unit 2, Department of Medical Oncology, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: A M
authors.initials: AM
authors.lastname: Gianni
authors.affiliation: Human Tumors Immunobiology Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Mortarini
authors.affiliation: Human Tumors Immunobiology Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, and Medical Oncology, Università degli Studi di Milano, Milan, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Anichini
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage*; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D001562:Benzimidazoles / Q000008:administration & dosage; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm; D004357:Drug Synergism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D008545:Melanoma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D051379:Mice; D016513:Mice, SCID; D009389:Neovascularization, Pathologic; D053218:Receptors, Death Domain / Q000235:genetics / Q000378:metabolism*; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000008:administration & dosage
chemical_list: C517975:AZD 6244; D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D001562:Benzimidazoles; D053218:Receptors, Death Domain; D053221:TNF-Related Apoptosis-Inducing Ligand