title: BIM upregulation and ROS-dependent necroptosis mediate the antitumor effects of the HDACi Givinostat and Sorafenib in Hodgkin lymphoma cell line xenografts.
abstract: Relapsed/refractory Hodgkin's lymphoma (HL) is an unmet medical need requiring new therapeutic options. Interactions between the histone deacetylase inhibitor Givinostat and the RAF/MEK/ERK inhibitor Sorafenib were examined in HDLM-2 and L-540 HL cell lines. Exposure to Givinostat/Sorafenib induced a synergistic inhibition of cell growth (range, 70-80%) and a marked increase in cell death (up to 96%) due to increased H3 and H4 acetylation and strong mitochondrial injury. Gene expression profiling indicated that the synergistic effects of Givinostat/Sorafenib treatment are associated with the modulation of cell cycle and cell death pathways. Exposure to Givinostat/Sorafenib resulted in sustained production of reactive oxygen species (ROS) and activation of necroptotic cell death. The necroptosis inhibitor Necrostatin-1 prevented Givinostat/Sorafenib-induced ROS production, mitochondrial injury, activation of BH3-only protein BIM and cell death. Knockdown experiments identified BIM as a key signaling molecule that mediates Givinostat/Sorafenib-induced oxidative death of HL cells. Furthermore, in vivo xenograft studies demonstrated a 50% reduction in tumor burden (P<0.0001), a 5- to 15-fold increase in BIM expression (P < 0.0001) and a fourfold increase in tumor necrosis in Givinostat/Sorafenib-treated animals compared with mice that received single agents. These results provide a rationale for exploring Givinostat/Sorafenib combination in relapsed/refractory HL.
authors.affiliation: 1] Department of Oncology and Hematology, Humanitas Cancer Center - Humanitas Clinical and Research Center, Milano, Italy [2] Department of Medical Biotechnology and Translational Medicine, University of Milano, Milano, Italy.
authors.forename: S L
authors.initials: SL
authors.lastname: Locatelli
authors.affiliation: Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale Tumori, Milano, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Cleris
authors.affiliation: 1] Department of Oncology and Hematology, Humanitas Cancer Center - Humanitas Clinical and Research Center, Milano, Italy [2] Department of Medical Biotechnology and Translational Medicine, University of Milano, Milano, Italy.
authors.forename: G G
authors.initials: GG
authors.lastname: Stirparo
authors.affiliation: Department of Oncology and Hematology, Humanitas Cancer Center - Humanitas Clinical and Research Center, Milano, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Tartari
authors.affiliation: Department of Oncology and Hematology, Humanitas Cancer Center - Humanitas Clinical and Research Center, Milano, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Saba
authors.affiliation: Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Pierdominici
authors.affiliation: 1] Department of Therapeutic Research and Medicine Evaluation, Istituto Superiore di Sanità, Rome, Italy [2] Istituto San Raffaele Sulmona, Sulmona, Italy.
authors.forename: W
authors.initials: W
authors.lastname: Malorni
authors.affiliation: Pathology Department, CRO Aviano, Aviano, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Carbone
authors.affiliation: Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale Tumori, Milano, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Anichini
authors.affiliation: 1] Department of Oncology and Hematology, Humanitas Cancer Center - Humanitas Clinical and Research Center, Milano, Italy [2] Department of Medical Biotechnology and Translational Medicine, University of Milano, Milano, Italy.
authors.forename: C
authors.identifier: 0000000331440124
authors.initials: C
authors.lastname: Carlo-Stella
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000502:physiology*; D000072224:Bcl-2-Like Protein 11; D002219:Carbamates / Q000008:administration & dosage*; D045744:Cell Line, Tumor; D056572:Histone Deacetylase Inhibitors / Q000008:administration & dosage*; D006689:Hodgkin Disease / Q000188:drug therapy* / Q000473:pathology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D051379:Mice; D016513:Mice, SCID; D009336:Necrosis; D009536:Niacinamide / Q000008:administration & dosage / Q000031:analogs & derivatives*; D010671:Phenylurea Compounds / Q000008:administration & dosage*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D000077157:Sorafenib; D015854:Up-Regulation; D023041:Xenograft Model Antitumor Assays
chemical_list: D051017:Apoptosis Regulatory Proteins; C000606775:BCL2L11 protein, human; D000072224:Bcl-2-Like Protein 11; C000606776:Bcl2l11 protein, mouse; D002219:Carbamates; D056572:Histone Deacetylase Inhibitors; D007093:Imidazoles; D007211:Indoles; D008565:Membrane Proteins; D010671:Phenylurea Compounds; D011518:Proto-Oncogene Proteins; D017382:Reactive Oxygen Species; C507699:necrostatin-1; D009536:Niacinamide; C575255:givinostat; D000077157:Sorafenib