The efficacy of nuclease-resistant Chol-siRNA in primary breast tumors following complexation with PLL-PEG(5K)

Vishakha V. Ambardekar, Rajesh R. Wakaskar, Bhawna Sharma, Joy Bowman, Willy Vayaboury, Rakesh K Singh, Joseph A Vetro

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Modifying the sense strand of nuclease-resistant siRNA with 3'-cholesterol (Chol-*siRNA) increases mRNA suppression after i.v. administration but with relatively low efficacy. We previously found evidence invitro that suggests complexation of Chol-siRNA with PLL-PEG(5K), a block copolymer of poly- l-lysine and 5kDa polyethylene glycol, may increase the efficacy of Chol-siRNA invivo in a PLL block length-dependent manner. In this study, the extent that polyplexes of PLL10-PEG(5K), PLL30-PEG(5K), and PLL50-PEG(5K) protect complexed Chol-siRNA in high concentrations of murine serum and affect the activity of Chol-*siRNA in murine 4T1 breast tumor epithelial cells invitro and in primary orthotopic tumors of 4T1 was compared. PLL-PEG(5K) required 3'-Chol to protect full-length siRNA from nuclease degradation in 90% (v/v) murine serum and protection was increased by increasing PLL block length and nuclease resistance of Chol-siRNA. Polyplexes of Chol-*siLuc suppressed stably expressed luciferase in 4T1-Luc cells to different levels invitro where PLL30>PLL50>PLL10. In contrast, only polyplexes of Chol-*siLuc and PLL30-PEG(5K) or PLL50-PEG(5K) suppressed high levels of luciferase in primary orthotopic tumors of 4T1-Luc after i.v. administration, whereas polyplexes of Chol-*siLuc and PLL10-PEG(5K), inactive Chol-*siCtrl polyplexes of PLL-PEG(5K), or Chol-*siLuc alone had no detectable activity. As a whole, these results indicate that polyplexes of PLL-PEG(5K) increase the efficacy of nuclease-resistant Chol-siRNA in primary breast tumors after i.v. administration in a PLL block length-dependent manner. Thus, complexation of Chol-siRNA with PLL-PEG(5K) may be a promising approach to increase the efficacy of Chol-siRNA in a wide range of primary tumors, metastases, and other tissues but likely requires a PLL block length that balances polymer-related adverse effects, Chol-siRNA bioavailability, and subsequent activity in the target cell.

Original languageEnglish (US)
Pages (from-to)4839-4848
Number of pages10
JournalBiomaterials
Volume34
Issue number20
DOIs
StatePublished - Jul 1 2013

Fingerprint

Phase locked loops
Complexation
Small Interfering RNA
Polyethylene glycols
Tumors
Breast Neoplasms
Luciferases
Neoplasms
Cholesterol
Serum
Biological Availability
Lysine
Block copolymers
Polymers
Epithelial Cells
Tissue
Neoplasm Metastasis
Degradation

Keywords

  • Drug delivery
  • Polymer siRNA complexes
  • RNA interference
  • RNAi
  • SiRNA delivery
  • SiRNA polyplexes

ASJC Scopus subject areas

  • Bioengineering
  • Ceramics and Composites
  • Biophysics
  • Biomaterials
  • Mechanics of Materials

Cite this

The efficacy of nuclease-resistant Chol-siRNA in primary breast tumors following complexation with PLL-PEG(5K). / Ambardekar, Vishakha V.; Wakaskar, Rajesh R.; Sharma, Bhawna; Bowman, Joy; Vayaboury, Willy; Singh, Rakesh K; Vetro, Joseph A.

In: Biomaterials, Vol. 34, No. 20, 01.07.2013, p. 4839-4848.

Research output: Contribution to journalArticle

Ambardekar, Vishakha V. ; Wakaskar, Rajesh R. ; Sharma, Bhawna ; Bowman, Joy ; Vayaboury, Willy ; Singh, Rakesh K ; Vetro, Joseph A. / The efficacy of nuclease-resistant Chol-siRNA in primary breast tumors following complexation with PLL-PEG(5K). In: Biomaterials. 2013 ; Vol. 34, No. 20. pp. 4839-4848.
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AU - Bowman, Joy

AU - Vayaboury, Willy

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