caa a22 4500 44531205X CHVBK 20180317142633.0 cr unu---uuuuu 170323e20110901xx s 000 0 eng 10.1007/s11095-011-0457-y doi (NATIONALLICENCE)springer-10.1007/s11095-011-0457-y Targeted Delivery to Neuroblastoma of Novel siRNA-anti-GD2-liposomes Prepared by Dual Asymmetric Centrifugation and Sterol-Based Post-Insertion Method [Elektronische Daten] [Joanna Adrian, Alexander Wolf, Annette Steinbach, Jochen Rössler, Regine Süss] ABSTRACT: Purpose: To optimise and simplify preparation of targeted liposomes for efficient siRNA delivery to neuroblastoma, the most common solid tumour in early childhood. Methods: Liposomes containing siRNA were prepared by combining the novel dual asymmetric centrifugation (DAC) method and the recently optimised sterol-based post-insertion technique (SPIT) to couple anti-GD2 antibody for selective interaction with neuroblastoma cells. Cultured human neuroblastoma cell lines were used to evaluate the efficiency of siRNA delivery. Results: The size of liposomes prepared by DAC ranged from 190 to 240nm; siRNA encapsulation efficiency was up to 50%. An average of 70 and 100 molecules of anti-GD2 antibody per particle were coupled. A significant association of liposomes with neuroblastoma cells as well as effective siRNA delivery was observed only when anti-GD2 antibody was coupled. Preliminary data suggest delivery of siRNA using anti-GD2-liposomes occurs via GD2-mediated endocytosis. Vascular endothelial growth factor A (VEGF-A) was down-regulated using siRNA delivered by anti-GD2-liposomes. Conclusions: DAC and SPIT allow for the straightforward preparation of liposomes for the targeted delivery of siRNA. Anti-GD2-liposomes thus produced can serve as versatile carriers of siRNA to neuroblastoma cells. Springer Science+Business Media, LLC, 2011 dual asymmetric centrifugation nationallicence GD2 antibody nationallicence neuroblastoma nationallicence post-insertion technique nationallicence siRNA delivery nationallicence Ab : antibody nationallicence Chol : cholesterol nationallicence cryo-TEM : cryo-transmission electron microscope nationallicence DAC : dual asymmetric centrifugation nationallicence DDAB : dimethyldioctadecylammonium (bromide salt) nationallicence DSPE-PEG2000 : 2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000] nationallicence FCS : fetal calf serum nationallicence GD2 : disialoganglioside nationallicence HN buffer : HEPES/NaCl buffer nationallicence HSPC : hydrogenated soybean phosphatidylcholine nationallicence L : liposomes nationallicence PBS : phosphate buffered saline nationallicence POPC : 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine nationallicence Rhd-PE : rhodamine B 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt nationallicence siRNA : short interfering RNA nationallicence SPIT : sterol-based post-insertion technique nationallicence TL : total lipid nationallicence VEGF-A : vascular endothelial growth factor A nationallicence Adrian Joanna Department of Pharmaceutical Technology & Biopharmacy, Albert-Ludwigs University, Sonnenstr. 5, 79104, Freiburg, Germany aut Wolf Alexander Center of Paediatrics & Adolescent Medicine Division of Paediatric Haematology & Oncology, University Medical Center Freiburg, Mathildenstr. 1, 79106, Freiburg, Germany aut Steinbach Annette Department of Pharmaceutical Technology & Biopharmacy, Albert-Ludwigs University, Sonnenstr. 5, 79104, Freiburg, Germany aut Rössler Jochen Center of Paediatrics & Adolescent Medicine Division of Paediatric Haematology & Oncology, University Medical Center Freiburg, Mathildenstr. 1, 79106, Freiburg, Germany aut Süss Regine Department of Pharmaceutical Technology & Biopharmacy, Albert-Ludwigs University, Sonnenstr. 5, 79104, Freiburg, Germany aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/9(2011-09-01), 2261-2272 0724-8741 28:9<2261 2011 28 11095 https://doi.org/10.1007/s11095-011-0457-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0457-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Adrian Joanna Department of Pharmaceutical Technology & Biopharmacy, Albert-Ludwigs University, Sonnenstr. 5, 79104, Freiburg, Germany aut NATIONALLICENCE 700 1- Wolf Alexander Center of Paediatrics & Adolescent Medicine Division of Paediatric Haematology & Oncology, University Medical Center Freiburg, Mathildenstr. 1, 79106, Freiburg, Germany aut NATIONALLICENCE 700 1- Steinbach Annette Department of Pharmaceutical Technology & Biopharmacy, Albert-Ludwigs University, Sonnenstr. 5, 79104, Freiburg, Germany aut NATIONALLICENCE 700 1- Rössler Jochen Center of Paediatrics & Adolescent Medicine Division of Paediatric Haematology & Oncology, University Medical Center Freiburg, Mathildenstr. 1, 79106, Freiburg, Germany aut NATIONALLICENCE 700 1- Süss Regine Department of Pharmaceutical Technology & Biopharmacy, Albert-Ludwigs University, Sonnenstr. 5, 79104, Freiburg, Germany aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/9(2011-09-01), 2261-2272 0724-8741 28:9<2261 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer