title: Selective lentiviral gene delivery to CD133-expressing human glioblastoma stem cells.
abstract: Glioblastoma multiforme (GBM) is a deadly primary brain malignancy. Glioblastoma stem cells (GSC), which have the ability to self-renew and differentiate into tumor lineages, are believed to cause tumor recurrence due to their resistance to current therapies. A subset of GSCs is marked by cell surface expression of CD133, a glycosylated pentaspan transmembrane protein. The study of CD133-expressing GSCs has been limited by the relative paucity of genetic tools that specifically target them. Here, we present CD133-LV, a lentiviral vector presenting a single chain antibody against CD133 on its envelope, as a vehicle for the selective transduction of CD133-expressing GSCs. We show that CD133-LV selectively transduces CD133+ human GSCs in dose-dependent manner and that transduced cells maintain their stem-like properties. The transduction efficiency of CD133-LV is reduced by an antibody that recognizes the same epitope on CD133 as the viral envelope and by shRNA-mediated knockdown of CD133. Conversely, the rate of transduction by CD133-LV is augmented by overexpression of CD133 in primary human GBM cultures. CD133-LV selectively transduces CD133-expressing cells in intracranial human GBM xenografts in NOD.SCID mice, but spares normal mouse brain tissue, neurons derived from human embryonic stem cells and primary human astrocytes. Our findings indicate that CD133-LV represents a novel tool for the selective genetic manipulation of CD133-expressing GSCs, and can be used to answer important questions about how these cells contribute to tumor biology and therapy resistance.
authors.affiliation: Department of Neurosurgery, NYU School of Medicine, New York, NY, United States of America; Helen L. and Martin S. Kimmel Center for Stem Cell Biology, NYU School of Medicine, New York, NY, United States of America.
authors.forename: N Sumru
authors.initials: NS
authors.lastname: Bayin
authors.affiliation: Department of Neurosurgery, NYU School of Medicine, New York, NY, United States of America; Medical Scientist Training Program, NYU School of Medicine, New York, NY, United States of America.
authors.forename: Aram S
authors.initials: AS
authors.lastname: Modrek
authors.affiliation: Department of Neurosurgery, NYU School of Medicine, New York, NY, United States of America.
authors.forename: August
authors.initials: A
authors.lastname: Dietrich
authors.affiliation: Department of Neurosurgery, NYU School of Medicine, New York, NY, United States of America.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Lebowitz
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Abel
authors.affiliation: Department of Neurosurgery, NYU School of Medicine, New York, NY, United States of America.
authors.forename: Hae-Ri
authors.initials: HR
authors.lastname: Song
authors.affiliation: Helen L. and Martin S. Kimmel Center for Stem Cell Biology, NYU School of Medicine, New York, NY, United States of America; Department of Cell Biology, NYU School of Medicine, New York, NY, United States of America; Ronald O. Perelman Department of Dermatology, NYU School of Medicine, New York, NY, United States of America.
authors.forename: Markus
authors.initials: M
authors.lastname: Schober
authors.affiliation: Department of Pathology, NYU School of Medicine, New York, NY, United States of America.
authors.forename: David
authors.initials: D
authors.lastname: Zagzag
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany; German Cancer Consortium, Heidelberg, Germany.
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Buchholz
authors.affiliation: Skirball Institute, NYU School of Medicine, New York, NY, United States of America.
authors.forename: Moses V
authors.initials: MV
authors.lastname: Chao
authors.affiliation: Department of Neurosurgery, NYU School of Medicine, New York, NY, United States of America; Helen L. and Martin S. Kimmel Center for Stem Cell Biology, NYU School of Medicine, New York, NY, United States of America; Brain Tumor Center, NYU School of Medicine, New York, NY, United States of America.
authors.forename: Dimitris G
authors.initials: DG
authors.lastname: Placantonakis
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000032:analysis / Q000276:immunology*; D001932:Brain Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology; D005909:Glioblastoma / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D006023:Glycoproteins / Q000032:analysis / Q000276:immunology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000276:immunology; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014411:Neoplastic Stem Cells / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D010455:Peptides / Q000032:analysis / Q000276:immunology*; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse