title: Pediatric medulloblastoma xenografts including molecular subgroup 3 and CD133+ and CD15+ cells are sensitive to killing by oncolytic herpes simplex viruses.
abstract: BACKGROUND
  Childhood medulloblastoma is associated with significant morbidity and mortality that is compounded by neurotoxicity for the developing brain caused by current therapies, including surgery, craniospinal radiation, and chemotherapy. Innate therapeutic resistance of some aggressive pediatric medulloblastoma has been attributed to a subpopulation of cells, termed cancer-initiating cells or cancer stemlike cells (CSCs), marked by the surface protein CD133 or CD15. Brain tumors characteristically contain areas of pathophysiologic hypoxia, which has been shown to drive the CSC phenotype leading to heightened invasiveness, angiogenesis, and metastasis. Novel therapies that target medulloblastoma CSCs are needed to improve outcomes and decrease toxicity. We hypothesized that oncolytic engineered herpes simplex virus (oHSV) therapy could effectively infect and kill pediatric medulloblastoma cells, including CSCs marked by CD133 or CD15.
  
  
  METHODS
  Using 4 human pediatric medulloblastoma xenografts, including 3 molecular subgroup 3 tumors, which portend worse patient outcomes, we determined the expression of CD133, CD15, and the primary HSV-1 entry molecule nectin-1 (CD111) by fluorescence activated cell sorting (FACS) analysis. Infectability and cytotoxicity of clinically relevant oHSVs (G207 and M002) were determined in vitro and in vivo by FACS, immunofluorescent staining, cytotoxicity assays, and murine survival studies.
  
  
  RESULTS
  We demonstrate that hypoxia increased the CD133+ cell fraction, while having the opposite effect on CD15 expression. We established that all 4 xenografts, including the CSCs, expressed CD111 and were highly sensitive to killing by G207 or M002.
  
  
  CONCLUSIONS
  Pediatric medulloblastoma, including Group 3 tumors, may be an excellent target for oHSV virotherapy, and a clinical trial in medulloblastoma is warranted.
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: Gregory K
authors.initials: GK
authors.lastname: Friedman
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: Blake P
authors.initials: BP
authors.lastname: Moore
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: Li
authors.initials: L
authors.lastname: Nan
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: Virginia M
authors.initials: VM
authors.lastname: Kelly
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: Tina
authors.initials: T
authors.lastname: Etminan
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: Catherine P
authors.initials: CP
authors.lastname: Langford
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: Hui
authors.initials: H
authors.lastname: Xu
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: Xiaosi
authors.initials: X
authors.lastname: Han
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: James M
authors.initials: JM
authors.lastname: Markert
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Beierle
authors.affiliation: Division of Pediatric Hematology and Oncology, Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, Alabama (G.K.F., B.P.M., L.N., V.M.K.); Science and Technology Honors Program, Department of Chemistry, University of Alabama at Birmingham, Birmingham, Alabama (T.E.); Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, Alabama (C.P.L., J.M.M., G.Y.G.); Department of Dermatology, University of Alabama at Birmingham, Birmingham, Alabama (H.X.); Department of Neurology, University of Alabama at Birmingham, Birmingham, Alabama (X.H.); Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama (E.A.B.).
authors.forename: G Yancey
authors.initials: GY
authors.lastname: Gillespie
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D017209:Apoptosis; D049109:Cell Proliferation; D002528:Cerebellar Neoplasms / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D002648:Child; D005647:Fucosyltransferases / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D016256:Lewis X Antigen / Q000378:metabolism*; D008527:Medulloblastoma / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D051379:Mice; D008819:Mice, Nude; D050130:Oncolytic Virotherapy; D050504:Oncolytic Viruses / Q000235:genetics*; D010455:Peptides / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014779:Virus Replication; D023041:Xenograft Model Antitumor Assays
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; D016256:Lewis X Antigen; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; C066693:FUT4 protein, human; D005647:Fucosyltransferases
keywords: CD133; CD15; HSV; medulloblastoma; virotherapy