title: Natural killer cell lines preferentially kill clonogenic multiple myeloma cells and decrease myeloma engraftment in a bioluminescent xenograft mouse model.
abstract: BACKGROUND
  Novel therapies capable of targeting drug resistant clonogenic MM cells are required for more effective treatment of multiple myeloma. This study investigates the cytotoxicity of natural killer cell lines against bulk and clonogenic multiple myeloma and evaluates the tumor burden after NK cell therapy in a bioluminescent xenograft mouse model.
  
  
  METHODS
  The cytotoxicity of natural killer cell lines was evaluated against bulk multiple myeloma cell lines using chromium release and flow cytometry cytotoxicity assays. Selected activating receptors on natural killer cells were blocked to determine their role in multiple myeloma recognition. Growth inhibition of clonogenic multiple myeloma cells was assessed in a methylcellulose clonogenic assay in combination with secondary replating to evaluate the self-renewal of residual progenitors after natural killer cell treatment. A bioluminescent mouse model was developed using the human U266 cell line transduced to express green fluorescent protein and luciferase (U266eGFPluc) to monitor disease progression in vivo and assess bone marrow engraftment after intravenous NK-92 cell therapy.
  
  
  RESULTS
  Three multiple myeloma cell lines were sensitive to NK-92 and KHYG-1 cytotoxicity mediated by NKp30, NKp46, NKG2D and DNAM-1 activating receptors. NK-92 and KHYG-1 demonstrated 2- to 3-fold greater inhibition of clonogenic multiple myeloma growth, compared with killing of the bulk tumor population. In addition, the residual colonies after treatment formed significantly fewer colonies compared to the control in a secondary replating for a cumulative clonogenic inhibition of 89-99% at the 20:1 effector to target ratio. Multiple myeloma tumor burden was reduced by NK-92 in a xenograft mouse model as measured by bioluminescence imaging and reduction in bone marrow engraftment of U266eGFPluc cells by flow cytometry.
  
  
  CONCLUSIONS
  This study demonstrates that NK-92 and KHYG-1 are capable of killing clonogenic and bulk multiple myeloma cells. In addition, multiple myeloma tumor burden in a xenograft mouse model was reduced by intravenous NK-92 cell therapy. Since multiple myeloma colony frequency correlates with survival, our observations have important clinical implications and suggest that clinical studies of NK cell lines to treat MM are warranted.
authors.affiliation: Cell Therapy Program, Princess Margaret Hospital, 610 University Ave, Toronto, ON, Canada.
authors.forename: Brenna E
authors.initials: BE
authors.lastname: Swift
authors.forename: Brent A
authors.initials: BA
authors.lastname: Williams
authors.forename: Yoko
authors.initials: Y
authors.lastname: Kosaka
authors.forename: Xing-Hua
authors.initials: XH
authors.lastname: Wang
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Medin
authors.forename: Sowmya
authors.initials: S
authors.lastname: Viswanathan
authors.forename: Joaquin
authors.initials: J
authors.lastname: Martinez-Lopez
authors.forename: Armand
authors.initials: A
authors.lastname: Keating
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002999:Clone Cells; D003602:Cytotoxicity, Immunologic; D005434:Flow Cytometry; D017930:Genes, Reporter; D006801:Humans; D007118:Immunoassay; D007155:Immunologic Factors / Q000008:administration & dosage; D016219:Immunotherapy, Adoptive; D007275:Injections, Intravenous; D007694:Killer Cells, Natural / Q000276:immunology* / Q000378:metabolism / Q000637:transplantation; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements; D051379:Mice; D009101:Multiple Myeloma / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D055607:Receptors, Natural Killer Cell / Q000037:antagonists & inhibitors / Q000276:immunology; D014183:Transplantation, Heterologous; D014410:Tumor Stem Cell Assay
chemical_list: D007155:Immunologic Factors; D055607:Receptors, Natural Killer Cell; D008156:Luciferases