Design of peptide-targeted liposomes containing nucleic acids

Adriana O. Santos, Lígia C. Gomes da Silva, Luís M. Bimbo, Maria C. Pedroso de Lima, Sérgio Simões, João N. Moreira

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Anticancer systemic gene silencing therapy has been so far limited by the inexistence of adequate carrier systems that ultimately provide an efficient intracellular delivery into target tumor cells. In this respect, one promising strategy involves the covalent attachment of internalizing-targeting ligands at the extremity of PEG chains grafted onto liposomes. Therefore, the present work aims at designing targeted liposomes containing nucleic acids, with small size, high encapsulation efficiency and able to be actively internalized by SCLC cells, using a hexapeptide (antagonist G) as a targeting ligand. For this purpose, the effect of the liposomal preparation method, loading material (ODN versus siRNA) and peptide-coupling procedure (direct coupling versus post-insertion) on each of the above-mentioned parameters was assessed. Post-insertion of DSPE-PEG-antagonist G conjugates into preformed liposomes herein named as stabilized lipid particles, resulted in targeted vesicles with a mean size of about 130 nm, encapsulation efficiency close to 100%, and a loading capacity of approximately 5 nmol siRNA/mumol of total lipid. In addition, the developed targeted vesicles showed increased internalization in SCLC cells, as well as in other tumor cells and HMEC-1 microvascular endothelial cells. The improved cellular association, however, did not correlate with enhanced downregulation of the target protein (Bcl-2) in SCLC cells. These results indicate that additional improvements need to be performed in the future, namely by ameliorating the access of the nucleic acids to the cytoplasm of the tumor cells following receptor-mediated endocytosis.

LanguageEnglish
Pages433-441
Number of pages9
JournalBBA - Biochimica et Biophysica Acta
Volume1798
Issue number3
Early online date11 Dec 2009
DOIs
Publication statusPublished - 31 Mar 2010
Externally publishedYes

Fingerprint

Liposomes
Nucleic Acids
Peptides
Small Interfering RNA
Ligands
Lipids
Neoplasms
Gene Silencing
Endocytosis
Genetic Therapy
Cytoplasm
Down-Regulation
Extremities
Endothelial Cells
Proteins
arginyl-tryptophyl-N-methylphenylalanyl-tryptophyl-leucyl-methioninamide

Keywords

  • cell line, tumor
  • drug delivery systems
  • endocytosis
  • fluorometry
  • humans
  • liposomes
  • nNucleic acids
  • peptides
  • proto-oncogene proteins c-bcl-2
  • RNA, small interfering
  • transfection

Cite this

Santos, A. O., da Silva, L. C. G., Bimbo, L. M., de Lima, M. C. P., Simões, S., & Moreira, J. N. (2010). Design of peptide-targeted liposomes containing nucleic acids. BBA - Biochimica et Biophysica Acta , 1798(3), 433-441. https://doi.org/10.1016/j.bbamem.2009.12.001
Santos, Adriana O. ; da Silva, Lígia C. Gomes ; Bimbo, Luís M. ; de Lima, Maria C. Pedroso ; Simões, Sérgio ; Moreira, João N. / Design of peptide-targeted liposomes containing nucleic acids. In: BBA - Biochimica et Biophysica Acta . 2010 ; Vol. 1798, No. 3. pp. 433-441.
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Santos, AO, da Silva, LCG, Bimbo, LM, de Lima, MCP, Simões, S & Moreira, JN 2010, 'Design of peptide-targeted liposomes containing nucleic acids' BBA - Biochimica et Biophysica Acta , vol. 1798, no. 3, pp. 433-441. https://doi.org/10.1016/j.bbamem.2009.12.001

Design of peptide-targeted liposomes containing nucleic acids. / Santos, Adriana O.; da Silva, Lígia C. Gomes; Bimbo, Luís M.; de Lima, Maria C. Pedroso; Simões, Sérgio; Moreira, João N.

In: BBA - Biochimica et Biophysica Acta , Vol. 1798, No. 3, 31.03.2010, p. 433-441.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Design of peptide-targeted liposomes containing nucleic acids

AU - Santos, Adriana O.

AU - da Silva, Lígia C. Gomes

AU - Bimbo, Luís M.

AU - de Lima, Maria C. Pedroso

AU - Simões, Sérgio

AU - Moreira, João N.

N1 - Copyright 2009 Elsevier B.V. All rights reserved.

PY - 2010/3/31

Y1 - 2010/3/31

N2 - Anticancer systemic gene silencing therapy has been so far limited by the inexistence of adequate carrier systems that ultimately provide an efficient intracellular delivery into target tumor cells. In this respect, one promising strategy involves the covalent attachment of internalizing-targeting ligands at the extremity of PEG chains grafted onto liposomes. Therefore, the present work aims at designing targeted liposomes containing nucleic acids, with small size, high encapsulation efficiency and able to be actively internalized by SCLC cells, using a hexapeptide (antagonist G) as a targeting ligand. For this purpose, the effect of the liposomal preparation method, loading material (ODN versus siRNA) and peptide-coupling procedure (direct coupling versus post-insertion) on each of the above-mentioned parameters was assessed. Post-insertion of DSPE-PEG-antagonist G conjugates into preformed liposomes herein named as stabilized lipid particles, resulted in targeted vesicles with a mean size of about 130 nm, encapsulation efficiency close to 100%, and a loading capacity of approximately 5 nmol siRNA/mumol of total lipid. In addition, the developed targeted vesicles showed increased internalization in SCLC cells, as well as in other tumor cells and HMEC-1 microvascular endothelial cells. The improved cellular association, however, did not correlate with enhanced downregulation of the target protein (Bcl-2) in SCLC cells. These results indicate that additional improvements need to be performed in the future, namely by ameliorating the access of the nucleic acids to the cytoplasm of the tumor cells following receptor-mediated endocytosis.

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KW - cell line, tumor

KW - drug delivery systems

KW - endocytosis

KW - fluorometry

KW - humans

KW - liposomes

KW - nNucleic acids

KW - peptides

KW - proto-oncogene proteins c-bcl-2

KW - RNA, small interfering

KW - transfection

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DO - 10.1016/j.bbamem.2009.12.001

M3 - Article

VL - 1798

SP - 433

EP - 441

JO - BBA - Biochimica et Biophysica Acta

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JF - BBA - Biochimica et Biophysica Acta

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