title: Targeted delivery to neuroblastoma of novel siRNA-anti-GD2-liposomes prepared by dual asymmetric centrifugation and sterol-based post-insertion method.
abstract: OBJECTIVE
  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 240 nm; 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.
authors: Department of Pharmaceutical Technology & Biopharmacy, Albert-Ludwigs University, Sonnenstr. 5, 79104 Freiburg, Germany. joanna.adrian@gmail.com
authors: Joanna E
authors: JE
authors: Adrian
authors: Alexander
authors: A
authors: Wolf
authors: Annette
authors: A
authors: Steinbach
authors: Jochen
authors: J
authors: Rössler
authors: Regine
authors: R
authors: Süss
references: J Control Release. 2010 Jun 15;144(3):341-9
references: 20226822
references: Cancer Biol Ther. 2009 May;8(10):874-82
references: 19342881
references: Cancer Res. 2003 Jul 15;63(14):3919-22
references: 12873985
references: Nature. 2001 May 24;411(6836):494-8
references: 11373684
references: Int J Cancer. 1997 Sep 26;73(1):42-9
references: 9334808
references: Methods Mol Biol. 2008;440:15-33
references: 18369934
references: Nature. 2006 May 4;441(7089):111-4
references: 16565705
references: J Pediatr Hematol Oncol. 1997 Sep-Oct;19(5):428-32
references: 9329464
references: J Control Release. 2008 Jan 4;125(1):16-24
references: 18023907
references: J Control Release. 2009 Apr 2;135(1):80-8
references: 19124051
references: EMBO Mol Med. 2009 Jun;1(3):142-51
references: 20049714
references: Pharm Res. 2009 Mar;26(3):529-38
references: 19015959
references: Clin Exp Metastasis. 2003;20(4):311-9
references: 12856718
references: Glycoconj J. 2004;20(5):301-17
references: 15229395
references: Cancer Res. 2008 Nov 1;68(21):9078-86
references: 18974154
references: Nat Rev Drug Discov. 2005 Feb;4(2):145-60
references: 15688077
references: Science. 2008 Feb 1;319(5863):627-30
references: 18239128
references: J Exp Clin Cancer Res. 2009 Nov 06;28:143
references: 19895696
references: Cancer Lett. 2005 Jul 8;225(1):141-9
references: 15922866
references: Biochim Biophys Acta. 2007 Jun;1768(6):1430-9
references: 17493581
references: Clin Cancer Res. 2006 Sep 1;12(17):5018-22
references: 16951216
references: Gene Ther. 2006 Mar;13(6):464-77
references: 16341059
references: Nat Rev Drug Discov. 2007 Jun;6(6):443-53
references: 17541417
references: Nat Rev Cancer. 2003 Mar;3(3):203-16
references: 12612655
references: J Biol Chem. 1959 Mar;234(3):466-8
references: 13641241