title: TGFβR1 Blockade with Galunisertib (LY2157299) Enhances Anti-Neuroblastoma Activity of the Anti-GD2 Antibody Dinutuximab (ch14.18) with Natural Killer Cells.
abstract: OBJECTIVE
  Immunotherapy of high-risk neuroblastoma using the anti-GD2 antibody dinutuximab induces antibody-dependent cell-mediated cytotoxicity (ADCC). Galunisertib, an inhibitor of TGFβR1, was examined for its ability to enhance the efficacy of dinutuximab in combination with human ex vivo activated NK (aNK) cells against neuroblastoma.
  
  
  METHODS
  TGFB1 and TGFBR1 mRNA expression was determined for 249 primary neuroblastoma tumors by microarray analysis. The ability of galunisertib to inhibit SMAD activity induced by neuroblastoma patient blood and bone marrow plasmas in neuroblastoma cells was tested. The impact of galunisertib on TGFβ1-induced inhibition of aNK cytotoxicity and ADCC in vitro and on anti-neuroblastoma activity in NOD-scid gamma (NSG) mice was determined.
  
  
  RESULTS
  Neuroblastomas express TGFB1 and TGFBR1 mRNA. Galunisertib suppressed SMAD activation in neuroblastoma cells induced by exogenous TGFβ1 or by patient blood and bone marrow plasma, and suppressed SMAD2 phosphorylation in human neuroblastoma cells growing in NSG mice. In NK cells treated in vitro with exogenous TGFβ1, galunisertib suppressed SMAD2 phosphorylation and restored the expression of DNAM-1, NKp30, and NKG2D cytotoxicity receptors and the TRAIL death ligand, the release of perforin and granzyme A, and the direct cytotoxicity and ADCC of aNK cells against neuroblastoma cells. Addition of galunisertib to adoptive cell therapy with aNK cells plus dinutuximab reduced tumor growth and increased survival of mice injected with two neuroblastoma cell lines or a patient-derived xenograft.
  
  
  CONCLUSIONS
  Galunisertib suppresses activation of SMAD2 in neuroblastomas and aNK cells, restores NK cytotoxic mechanisms, and increases the efficacy of dinutuximab with aNK cells against neuroblastoma tumors. Clin Cancer Res; 23(3); 804-13. ©2016 AACRSee related commentary by Zenarruzabeitia et al., p. 615.
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California.|Department of Pediatrics, Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Hung C
authors.initials: HC
authors.lastname: Tran
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California.
authors.forename: Zesheng
authors.initials: Z
authors.lastname: Wan
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Sheard
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California.
authors.forename: Jianping
authors.initials: J
authors.lastname: Sun
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California.|Department of Surgery, Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Jeremy R
authors.initials: JR
authors.lastname: Jackson
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California.
authors.forename: Jemily
authors.initials: J
authors.lastname: Malvar
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California.
authors.forename: Yibing
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Pathology and Laboratory Medicine, Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Larry
authors.initials: L
authors.lastname: Wang
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California.|Department of Pediatrics, Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Richard
authors.initials: R
authors.lastname: Sposto
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California.|Department of Surgery, Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Eugene S
authors.initials: ES
authors.lastname: Kim
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California.|Department of Pediatrics, Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Shahab
authors.initials: S
authors.lastname: Asgharzadeh
authors.affiliation: Children's Hospital Los Angeles and the Saban Research Institute, Los Angeles, California. rseeger@chla.usc.edu.|Department of Pediatrics, Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Seeger
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000074322:Antineoplastic Agents, Immunological / Q000494:pharmacology; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic; D004357:Drug Synergism; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D016219:Immunotherapy, Adoptive; D007694:Killer Cells, Natural / Q000637:transplantation*; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000502:physiology; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology*; D010766:Phosphorylation / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D011720:Pyrazoles / Q000494:pharmacology*; D011804:Quinolines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012334:RNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D051899:Smad2 Protein / Q000037:antagonists & inhibitors / Q000378:metabolism; D013047:Specific Pathogen-Free Organisms; D053773:Transforming Growth Factor beta1 / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D023041:Xenograft Model Antitumor Assays
chemical_list: D000911:Antibodies, Monoclonal; D000074322:Antineoplastic Agents, Immunological; D009363:Neoplasm Proteins; D011720:Pyrazoles; D011804:Quinolines; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D018125:Receptors, Transforming Growth Factor beta; C497782:SMAD2 protein, human; D051899:Smad2 Protein; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; C557799:LY-2157299; C112746:dinutuximab; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625841:TGFBR1 protein, human; C000625842:Tgfbr1 protein, mouse