title: Development of a Sox2 reporter system modeling cellular heterogeneity in glioma.
abstract: BACKGROUND
  Malignant gliomas are complex systems containing a number of factors that drive tumor initiation and progression, including genetic aberrations that lead to extensive cellular heterogeneity within the neoplastic compartment. Mouse models recapitulate these genetic aberrations, but readily observable heterogeneity remains challenging.
  
  
  METHODS
  To interrogate cellular heterogeneity in mouse glioma models, we utilized a replication-competent avian sarcoma-leukosis virus long terminal repeat with splice acceptor/tumor virus A (RCAS-tva) system to generate spontaneous mouse gliomas that contained a Sox2-enhanced green fluorescent protein (EGFP) reporter. Glial fibrillary acidic protein-tva mice were crossed with Sox2-EGFP mice, and tumors were initiated that contained a subpopulation of Sox2-EGFP-high cells enriched for tumor-initiating cell properties such as self-renewal, multilineage differentiation potential, and perivascular localization.
  
  
  RESULTS
  Following implantation into recipient mice, Sox2-EGFP-high cells generated tumors containing Sox2-EGFP-high and Sox2-EGFP-low cells. Kinomic analysis of Sox2-EGFP-high cells revealed activation of known glioma signaling pathways that are strongly correlated with patient survival including platelet-derived growth factor receptor beta, phosphoinositide-3 kinase, and vascular endothelial growth factor. Our functional analysis identified active feline sarcoma (Fes) signaling in Sox2-EGFP-high cells. Fes negatively correlated with glioma patient survival and was coexpressed with Sox2-positive cells in glioma xenografts and primary patient-derived tissue.
  
  
  CONCLUSIONS
  Our RCAS-tva/Sox2-EGFP model will empower closer examination of cellular heterogeneity and will be useful for identifying novel glioma pathways as well as testing preclinical treatment efficacy.
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (K.S., M.S., J.S.H., B.O., J.D.L.); Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (Q.W., J.N.R.); Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, Ohio (M.S., J.D.L.); Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama (K.W., A.B.H.); Image I.Q., Cleveland, Ohio (A.V.); Case Comprehensive Cancer Center, Cleveland, Ohio (J.N.R., J.D.L.).
authors.forename: Kevin
authors.initials: K
authors.lastname: Stoltz
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (K.S., M.S., J.S.H., B.O., J.D.L.); Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (Q.W., J.N.R.); Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, Ohio (M.S., J.D.L.); Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama (K.W., A.B.H.); Image I.Q., Cleveland, Ohio (A.V.); Case Comprehensive Cancer Center, Cleveland, Ohio (J.N.R., J.D.L.).
authors.forename: Maksim
authors.initials: M
authors.lastname: Sinyuk
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (K.S., M.S., J.S.H., B.O., J.D.L.); Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (Q.W., J.N.R.); Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, Ohio (M.S., J.D.L.); Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama (K.W., A.B.H.); Image I.Q., Cleveland, Ohio (A.V.); Case Comprehensive Cancer Center, Cleveland, Ohio (J.N.R., J.D.L.).
authors.forename: James S
authors.initials: JS
authors.lastname: Hale
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (K.S., M.S., J.S.H., B.O., J.D.L.); Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (Q.W., J.N.R.); Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, Ohio (M.S., J.D.L.); Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama (K.W., A.B.H.); Image I.Q., Cleveland, Ohio (A.V.); Case Comprehensive Cancer Center, Cleveland, Ohio (J.N.R., J.D.L.).
authors.forename: Qiulian
authors.initials: Q
authors.lastname: Wu
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (K.S., M.S., J.S.H., B.O., J.D.L.); Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (Q.W., J.N.R.); Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, Ohio (M.S., J.D.L.); Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama (K.W., A.B.H.); Image I.Q., Cleveland, Ohio (A.V.); Case Comprehensive Cancer Center, Cleveland, Ohio (J.N.R., J.D.L.).
authors.forename: Balint
authors.initials: B
authors.lastname: Otvos
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (K.S., M.S., J.S.H., B.O., J.D.L.); Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (Q.W., J.N.R.); Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, Ohio (M.S., J.D.L.); Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama (K.W., A.B.H.); Image I.Q., Cleveland, Ohio (A.V.); Case Comprehensive Cancer Center, Cleveland, Ohio (J.N.R., J.D.L.).
authors.forename: Kiera
authors.initials: K
authors.lastname: Walker
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (K.S., M.S., J.S.H., B.O., J.D.L.); Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (Q.W., J.N.R.); Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, Ohio (M.S., J.D.L.); Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama (K.W., A.B.H.); Image I.Q., Cleveland, Ohio (A.V.); Case Comprehensive Cancer Center, Cleveland, Ohio (J.N.R., J.D.L.).
authors.forename: Amit
authors.initials: A
authors.lastname: Vasanji
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (K.S., M.S., J.S.H., B.O., J.D.L.); Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (Q.W., J.N.R.); Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, Ohio (M.S., J.D.L.); Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama (K.W., A.B.H.); Image I.Q., Cleveland, Ohio (A.V.); Case Comprehensive Cancer Center, Cleveland, Ohio (J.N.R., J.D.L.).
authors.forename: Jeremy N
authors.initials: JN
authors.lastname: Rich
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (K.S., M.S., J.S.H., B.O., J.D.L.); Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (Q.W., J.N.R.); Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, Ohio (M.S., J.D.L.); Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama (K.W., A.B.H.); Image I.Q., Cleveland, Ohio (A.V.); Case Comprehensive Cancer Center, Cleveland, Ohio (J.N.R., J.D.L.).
authors.forename: Anita B
authors.initials: AB
authors.lastname: Hjelmeland
authors.affiliation: Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (K.S., M.S., J.S.H., B.O., J.D.L.); Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio (Q.W., J.N.R.); Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, Ohio (M.S., J.D.L.); Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama (K.W., A.B.H.); Image I.Q., Cleveland, Ohio (A.V.); Case Comprehensive Cancer Center, Cleveland, Ohio (J.N.R., J.D.L.).
authors.forename: Justin D
authors.initials: JD
authors.lastname: Lathia
mesh_terms: D000818:Animals; D001354:Avian Leukosis Virus / Q000235:genetics; D001358:Avian Sarcoma Viruses / Q000235:genetics; D001932:Brain Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology* / Q000821:virology; D004195:Disease Models, Animal; D017930:Genes, Reporter; D005822:Genetic Vectors; D005910:Glioma / Q000235:genetics / Q000378:metabolism / Q000473:pathology* / Q000821:virology; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology*; D055748:SOXB1 Transcription Factors / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D014407:Tumor Cells, Cultured
chemical_list: C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: RCAS/tva; Sox2; glioma; intratumoral heterogeneity