title: Targeting by cmHsp70.1-antibody coated and survivin miRNA plasmid loaded nanoparticles to radiosensitize glioblastoma cells.
abstract: Nanoparticles (NP) as carriers for anti-cancer drugs have shown great promise. Specific targeting of NP to malignant cells, however, remains an unsolved problem. Conjugation of antibodies specific for tumor membrane antigens to NP represents one approach to improve specificity and to increase therapeutic efficacy. In the present study, for the first time a novel membrane heat shock protein (Hsp70)-specific antibody (cmHsp70.1) was coupled to human serum albumin (HSA) NP, loaded with microRNA (miRNA) plasmids to target the inhibitor of apoptosis protein survivin. The physicochemical properties of monodisperse miRNA-loaded NP showed a diameter of 180 nm to 220 nm, a plasmid incorporation of more than 95% and a surface binding capacity of the antibody of 70-80%. Antibody-conjugated NP displayed an increased cellular uptake in U87MG and LN229 glioblastoma cells compared to isotype control antibody, PEG-coupled controls and peripheral blood lymphocytes (PBL). Survivin expression was significantly reduced in cells treated with the Hsp70-miRNA-NP as compared to non-conjugated NP. Hsp70-miRNA-NP enhanced radiation-induced increase in caspase 3/7 activity and decrease in clonogenic cell survival. In summary, cmHsp70.1 miRNA-NP comprise an enhanced tumor cell uptake and increased therapeutic efficacy of radiation therapy in vitro and provide the basis for the development of antibody-based advanced carrier systems for a tumor cell specific targeting.
authors.affiliation: Department of Pharmaceutical Technology, University of Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt am Main, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Gaca
authors.affiliation: Department of Radiotherapy and Oncology, University Hospital Frankfurt am Main, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Reichert
authors.affiliation: Department of Radiation Oncology, Technische Universität München, Ismaninger Str. 22, 81675 München, Germany; CCG "Innate Immunity in Tumor Biology", Helmholtz Zentrum München (HMGU), Munich, Germany.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Multhoff
authors.affiliation: Department of Pharmaceutical Technology, University of Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt am Main, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Wacker
authors.affiliation: Department of Radiotherapy and Oncology, University Hospital Frankfurt am Main, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Hehlgans
authors.affiliation: Department of Radiation Oncology, Technische Universität München, Ismaninger Str. 22, 81675 München, Germany.
authors.forename: Claus
authors.initials: C
authors.lastname: Botzler
authors.affiliation: Department of Radiation Oncology, Technische Universität München, Ismaninger Str. 22, 81675 München, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Gehrmann
authors.affiliation: Department of Radiotherapy and Oncology, University Hospital Frankfurt am Main, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany.
authors.forename: Claus
authors.initials: C
authors.lastname: Rödel
authors.affiliation: Department of Pharmaceutical Technology, University of Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt am Main, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Kreuter
authors.affiliation: Department of Radiotherapy and Oncology, University Hospital Frankfurt am Main, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany. Electronic address: franz.roedel@kgu.de.
authors.forename: Franz
authors.initials: F
authors.lastname: Rödel
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry* / Q000276:immunology; D045744:Cell Line, Tumor; D055785:Gene Knockdown Techniques; D015316:Genetic Therapy; D005909:Glioblastoma / Q000235:genetics* / Q000473:pathology / Q000532:radiotherapy / Q000628:therapy*; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology*; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics*; D035683:MicroRNAs / Q000008:administration & dosage* / Q000235:genetics / Q000627:therapeutic use; D053758:Nanoparticles / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics / Q000627:therapeutic use; D000077022:Survivin
chemical_list: D055910:Antibodies, Immobilized; C107489:BIRC5 protein, human; D018840:HSP70 Heat-Shock Proteins; D051034:Inhibitor of Apoptosis Proteins; D035683:MicroRNAs; D000077022:Survivin
keywords: Expression plasmids; Glioblastoma; HSA nanoparticles; RNA interference; Radiosensitization; Survivin