Pharmaceutical Research

, 28:2261

Targeted Delivery to Neuroblastoma of Novel siRNA-anti-GD2-liposomes Prepared by Dual Asymmetric Centrifugation and Sterol-Based Post-Insertion Method

  • Joanna E. Adrian
  • Alexander Wolf
  • Annette Steinbach
  • Jochen Rössler
  • Regine Süss
Research Paper

DOI: 10.1007/s11095-011-0457-y

Cite this article as:
Adrian, J.E., Wolf, A., Steinbach, A. et al. Pharm Res (2011) 28: 2261. doi:10.1007/s11095-011-0457-y

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 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.

KEY WORDS

dual asymmetric centrifugationGD2 antibodyneuroblastomapost-insertion techniquesiRNA delivery

ABBREVIATIONS

Ab

antibody

Chol

cholesterol

cryo-TEM

cryo-transmission electron microscope

DAC

dual asymmetric centrifugation

DDAB

dimethyldioctadecylammonium (bromide salt)

DSPE-PEG2000

2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000]

FCS

fetal calf serum

GD2

disialoganglioside

HN buffer

HEPES/NaCl buffer

HSPC

hydrogenated soybean phosphatidylcholine

L

liposomes

PBS

phosphate buffered saline

POPC

1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine

Rhd-PE

rhodamine B 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt

siRNA

short interfering RNA

SPIT

sterol-based post-insertion technique

TL

total lipid

VEGF-A

vascular endothelial growth factor A

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Joanna E. Adrian
    • 1
    • 2
  • Alexander Wolf
    • 2
  • Annette Steinbach
    • 1
  • Jochen Rössler
    • 2
  • Regine Süss
    • 1
  1. 1.Department of Pharmaceutical Technology & BiopharmacyAlbert-Ludwigs UniversityFreiburgGermany
  2. 2.Center of Paediatrics & Adolescent Medicine Division of Paediatric Haematology & OncologyUniversity Medical Center FreiburgFreiburgGermany